MX2008002044A - Microorganisms with increased efficiency for methionine synthesis - Google Patents

Abstract

The present invention concerns methods for the production of microorganisms with increased efficiency for methionine synthesis. The present invention also concerns microorganisms with increased efficiency for methionine synthesis. Furthermore, the present invention concerns methods for determining the optimal metabolic flux for organisms with respect to methionine synthesis.

Description

MICROORGANISMS WITH INCREASED EFFICIENCY FOR SYNTHESIS OF METIONININE FIELD OF THE INVENTION The invention lies in the field of fine chemicals that are produced by organisms. Particularly, the present invention concerns methods for the production of microorganisms with increased efficiency for methionine synthesis. The present invention also concerns microorganisms with increased efficiency for methionine synthesis. Additionally, the present invention concerns methods to determine the optimal metabolic flux for organisms with respect to methionine synthesis.

Technological background Amino acids are used for different purposes, one field of application is the use as additives for food in the feed of humans and animals. Methionine is an essential amino acid that must be ingested with food. In addition to being essential for protein biosynthesis, methionine serves as a precursor for different metabolites such as glutathione, S-adenosyl methionine and biotin. It also acts as a donor of methyl groups in various cellular processes. Currently, the world annual production of methionine is around 500,000 tons. Methionine is the first limiting amino acid to feed avian livestock and because of this, it is mainly applied as a dietary supplement. In contrast to other industrial amino acids, methionine is applied almost exclusively as a racemate produced by chemical synthesis (DE 190 64 05). Since animals can metabolize both stereoisomers of methionine, direct feeding of the chemically produced racemic mixture is possible (D 'Mello and Lewis (1978) Effect of Nutrí tíon Deficiencies in Animáis: Amíno Acids, Rechgigl (Ed.) CRC Handbook Series in Nutrition and Food, 441-490). However, there is still a great interest in replacing existing chemical production with a biotechnological process. This is due to the fact that, at lower levels of supplementation, L-methionine is a better source of sulfur amino acids than D-methionine (Katz &Baker, (1975) Poul T. Sci., 545, 1667- 74). Moreover, the chemical process uses rather dangerous chemicals and produces substantial waste streams. An efficient biotechnological process can avoid all these disadvantages of chemical production. For other amino acids such as glutamate, lysine, threonine and tryptophan, it has been known to produce them using fermentation methods. For these purposes, certain microorganisms such as Escheri chia coli (E. coli) and Coryneba cteri um glummicum (C. gl utami cum) have proved to be particularly suitable. The production of amino acids by fermentation also has the particular advantage that only L-amino acids are produced and that environmentally problematic chemicals such as solvents, etc. which are used in chemical synthesis, are avoided. However, the fermentative production of methionine by microorganisms will only be an alternative to chemical synthesis if it allows the production of methionine on a commercial scale at a price comparable to that of chemical production. In the past, there have been attempts to use microorganisms such as E. coli and C. gl utami cum for the production of sulfur-containing compounds that are also commonly referred to as fine chemicals. These methods included classical selection of strains by mutagenesis as well as optimization of culture conditions, for example targeting, oxygen supply, composition of culture media, etc. (Kumar et al. (2005) Biotechnology Advances, 23, 41-61). One of the reasons why the fermentative production of methionine in microorganisms has not proven to be economically interesting probably results from the peculiarity of the biosynthesis and metabolic pathways that lead to methionine. In general, the basic metabolic pathways that lead to methionine synthesis in organisms such as E. coli and C. glutamicum are well known (eg Voet and Voet (1995) Biochemistry, 2nd edition, Jon Wiley & amp;; Sons, Inc and http://www.genome.jp/kegg/metabolism.html). However, the details of the methionine biosynthesis in C. glutamicum and E. coli undergo intensive research and have been recently reviewed in R? ckert et al. (Rückert et al. (2003), J. of Biotechnology, 104, 213-228) and Lee et al. (Lee et al (2003), Appl Microbiol Biotechnol, 62, 459-467). A key step in methionine biosynthesis is the incorporation of sulfur in the main carbon chain. The source of sulfur is regularly sulphate and must be absorbed by microorganisms. The microorganisms must then activate and reduce the sulfate. These steps require an energy input of 7 mol of ATP and 8 mol of NADPH per methionine molecule (Neidhardt et al. (1990) Physiology of the bacterial cell: a molecular approa ch, Sunderland, Massachusetts, USA, Sinauer Associates, Inc. .) In this way, methionine is the amino acid with respect to which a cell must provide most of the energy. As a consequence of this, the microorganisms that produce methionine have developed metabolic pathways that are under strict control with respect to the rate and amount of methionine synthesis (Neidhardt FC (1996) E. coli and S. typhimuri um, ASM Press Washington) . These regulatory mechanisms include, for example, feedback control mechanisms, ie, the metabolic pathways that produce methionine are sub-regulated with respect to their activity once the cell has produced sufficient quantities of methionine. Prior art methodologies for obtaining microorganisms that can be used for industrial-scale production of methionine by microorganisms focused primarily on overcoming the control mechanisms mentioned in the foregoing by identifying genes that are involved in methionine biosynthesis. These genes were then overexpressed or repressed, depending on their respective function with the ultimate goal of increasing the amount of methionine produced. In this context, the amount of methionine has been defined as the amount of methionine obtained by amount of cell mass or as the amount of methionine obtained by time and volume (space-time-yield) or as a combination of both factors ie, cell mass and space-time-performance. For example, WO 02/10209 describes the overexpression or repression of certain genes in order to increase the amount of methionine produced. Recently, Rey et al. (Rey et al. (2003), J. Biotechnol., 103, 51-65,) identified the McbR transcriptional repressor that controls the expression of genes involved in methionine biosynthesis such as met Y (which encodes O-acetyl-L). -homoserinsulfhydrilase), metK (which encodes S-adenosyl-methionine synthetase), hom (which encodes hydrosensitrogenase), cysK (which encodes L-cysteine synthase), cysl (which encodes NADPH-dependent sulfite reductase) and ssuD (which encodes alcansulfonate monooxygenase). Even though these approaches allowed the construction of strains of microorganisms that produced more methionine compared to the wild type with the amount of methionine being calculated by cell mass or by time and volume (space-time yield), an organism has not been described so far. industrially competitive methionine overproducer (Mondal et al (1996) Folia Microbiol. (Praha), 416, 465-72, (Kumar et al. (2005) Biotechnology Advances, 23, 41-61).

SUMMARY OF THE INVENTION It has been found that the amount of methionine produced by an organism that is typically calculated as the amount of methionine per kilogram of cell mass or by time and volume is not a sufficient indicator as to whether a methionine producing organism can be considered as an economically interesting and commercially viable alternative to the chemical production of this amino acid. Instead, in order to be an economically interesting alternative to the chemical synthesis method, a high-efficiency methionine-producing organism is required, that is, an organism that makes possible a high space-time performance of methionine over the basis of the energy input of the production system that can be represented by the amount or contribution of a carbon source such as glucose that is being consumed for the production of methionine. Thus, when deciding whether a methionine-producing organism can be considered as an alternative to chemical synthesis, the key parameter should not be the amount of methionine produced by weight of cell mass, but the efficiency, that is, the molar amount of methionine produced by the amount of energy input consumed by the system, for example in the form of glucose. In this context, it has also been found that in order to produce methionine at a high efficiency in a microorganism, the metabolic pathways of the organism that directly or indirectly contribute to the methionine synthesis must be used in an optimum manner with respect to the synthesis of methionine In this way, for an efficient production of methionine by an organism, the metabolic flow through the metabolic pathways must be modified. Modification may not only be required for those pathways that are directly involved in the synthesis of the methionine backbone, but also those pathways that provide additional substrates such as sulfur atoms in different oxidative states, nitrogen in the reduced state such as ammonia. , additional carbon precursors including Cl carbon sources such as serine, glycine and formate, methionine precursors and different tetrathydrofolate metabolites that are substituted with carbon in N5 and NIO. In addition, the energy for example in the form of reduction equivalents such as NADH, NADPH, FADH2 can be involved in the pathways leading to methionine. In this way, a microorganism that produces methionine very efficiently may require a high metabolic flow through the pathways that lead to the construction of methionine and provide precursors of it, but may require only low metabolic fluxes through pathways of biosynthesis for example of other amino acids. It is therefore an object of the present invention to identify the optimal metabolic flux through the pathways involved directly or indirectly in the synthesis of methionine in order to identify potential organisms that can be very efficient in methionine synthesis. A further object of the present invention is to provide methods for predicting the ideal metabolic flow through the various metabolic pathways of an organism for methionine synthesis in order to achieve an efficient methionine biosynthesis. A further object of the present invention is to provide methods for obtaining organisms that have an increased efficiency in methionine synthesis. The present invention is also directed to organisms that are more efficient with respect to the methionine synthesis. These and other objects, since they will become apparent from the subsequent description, are resolved by the subject as defined in the independent claims. The dependent claims relate to some of the modalities contemplated by the invention. In the course of the present invention an analysis of metabolic pathways, also referred to as elemental flow mode analysis or extreme pathway analysis, was used to study the metabolic properties of organisms with respect to the methionine synthesis. While previous metabolic pathway analysis has been described in the prior art for other cellular systems (Papin et al. (2004) Trends Biotechnol. 228, 400-405; Schilling et al. (2000) J. T eor. Biol, 2033, 229-248; Schuster et al. (1999) Trends Biotechnol. 172, 53-60), this type of analysis has not been considered with respect to efficiency of methionine production in organisms such as C. gl utamicum and E. coli The analysis of metabolic pathways commonly allows the calculation of a solution space that contains all possible distributions of equilibrium flow of a metabolic network. Therefore, the stoichiometry of the metabolic network studied, including energy, precursors as well as co-factors requirements are considered completely. In the present invention, this analysis in elementary flow mode was carried out for the first time with respect to the efficiency of methionine production when comparing the metabolic networks of the main industrial producers of amino acids such as C. glutamicum and E. coli For this purpose, models of biochemical reactions were constructed for C. gl utamicum and E. coli (see in the following). The models included all the relevant routes of sulfur metabolism that involve all the routes linked to methionine production. These models were constructed from the current biochemical knowledge of the organisms investigated (see below). Based on these models, the optimal metabolic flux through the various pathways was calculated in order to predict which pathways should be used more or less intensively in order to increase the efficiency of methionine production. In calculating these models, a model organism was obtained which, for a given set of conditions including the presence of external metabolites such as the carbon source and the sulfur source, may be optimal for methionine production. The present invention in this way concerns a method for designing an organism with increased efficiency for methionine synthesis. This method comprises the steps of describing or assigning parameters to an initial organism synthesizing methionine by means of a plurality of parameters, which are obtained on the basis of metabolic pathways already known in relation to methionine synthesis and which are related to the metabolic flux through the reaction of these pathways, and then determining an organism with increased efficiency for methionine synthesis by modifying at least one of the plurality of parameters and / or introducing at least one additional parameter such that the Methionine synthesis efficiency compared to the efficiency of methionine synthesis of the initial methionine synthesizer organism. Using this method, in this way it is possible to predict a theoretical organism that must allow efficiency in synthesis of methionine. The detailed execution of the method is described later. For the purposes of the invention, these parameters were defined in relation to the individual reactions of the metabolic network considered. In this way, the parameters for optimization were defined in relation to the existence of a reaction in the organism used, the stoichiometry of a reaction and the reversibility of the reaction. As a consequence, the parameters are related to the metabolic flux through the various reactions of the network. The present invention also relates to a device for designing an initial organism with increased efficiency for methionine synthesis, the device comprises a processor adapted to carry out the steps of the aforementioned method to predict optimized pathways for an organism with increased synthesis of methionine. The invention also relates to a computer readable medium in which a computer program for designing an organism with increased efficiency for methionine synthesis is stored. The means that can be read by computer that when being executed by a processor is adapted to carry out the steps of the method mentioned in the above to design a theoretically optimized organism with increased efficiency of methionine synthesis. The invention further relates to a program element for designing an organism with increased efficiency for methionine synthesis which, when it is being executed by a processor, it is adapted to carry out the steps of the method mentioned in the above. The invention also relates to methods for producing organisms with increased efficiency of methionine synthesis which make use of the foregoing predictions when genetically modifying a wild-type organism in order to influence the metabolic flux of that organism so that it is seems more like the forecasts of the methods mentioned in the above. This can be achieved by genetically modifying the organism so that the metabolic flux through a certain reaction path increases and / or decreases. Genetic modifications can be introduced by recombinant DNA technology. Furthermore this can also be achieved by other techniques such as but not limited to mutation and selection processes such as chemical or UV mutagenesis and subsequent selection by growth in media containing analogs of substrates, which leads to resistant strains with improved characteristics. The invention also relates to methods for producing organisms with increased efficiency of methionine synthesis that make use of the foregoing predictions when genetically modifying an organism that is not a wild-type organism, but that has already been genetically modified before, preferably to produce methionine in an increased yield of mass and / or time-space. Such organisms may be organisms that are known as methionine overproducers and include for example organisms in which genes for sulfate uptake, genes for cysteine biosynthesis and genes for methionine synthesis as well as genes for conversion of oxaloacetate to aspartate semialdehyde are overexpressed. In such organisms which have already been genetically modified the predictions mentioned in the above regarding increased efficiency of methionine synthesis can be implemented in order to influence the metabolic flux of that organism so that it more closely resembles the predictions of the methods mentioned. in the above. This can be achieved by genetically modifying the organism so that the metabolic flux through a certain reaction path increases and / or decreases. Genetic modifications can be introduced by recombinant DNA technology. Furthermore this can also be achieved by other techniques such as but not limited to mutation and selection processes such as chemical or UV mutagenesis and subsequent selection by growth in media containing analogs of substrates, which leads to resistant strains with improved characteristics. It has surprisingly been found that the theoretical predictions obtained with respect to a wild-type organism can be used to increase the efficiency of methionine synthesis also in an organism that already carries mutations for example in pathways related to methionine synthesis or example accessory ways that are related to them. In this way, it does not seem necessary that the theoretical forecasts are calculated on the basis of the respective starting organism but that the theoretical predictions obtained for a wild-type organism may be sufficient. However, the present invention certainly also considers a mode in which an optimal metabolic flow is calculated on the basis of an initial organism that already provides some of the mutations mentioned in the foregoing so that the prognostics can be used to genetically modify the organism. Particularly, the present invention relates to methods for producing microorganisms of the genus Corynebacterium um and Escherichia with increased efficiency of methionine production comprising the steps of increasing and / or introducing metabolic flux through pathways that have been used to construct the model mentioned in the above. These methods may additionally include the steps of at least partially lowering the metabolic flux through the aforementioned pathways. The present invention also relates to organisms with an increased efficiency of methionine synthesis that can be obtained by any of the methods mentioned in the above.

In addition, the present invention relates to the use of such an organism to produce methionine and to methods for producing methionine by culturing the organisms mentioned above and isolating methionine.

BRIEF DESCRIPTION OF THE FIGURES Figure 1 shows a network of stoichiometric reactions of a wild type organism C. gl utamicum that was used for elementary flow mode analysis. Figure 2 shows the analysis of metabolic pathways of C. gl utamicum and E. coli for methionine synthesis. Figure 3 shows the metabolic flux distribution of a wild type organism C. glutamicum with maximum theoretical yield of methionine. Figure 4 shows the metabolic flux distribution of a wild type E. coli organism with maximum theoretical methionine yield. Figure 5 shows the analysis of metabolic pathways of C. gl utamicum for synthesis of methionine with different carbon and sulfur sources. Figures 6 to 9 show various vectors which are used in the examples of the modalities. Figure 10 shows an optimized metabolic flux distribution of a strain of C. glutamicum in which additional metabolic pathways have been included.

DETAILED DESCRIPTION OF THE PRESENT INVENTION Before describing in detail how the method mentioned above can be carried out in order to identify a theoretical optimized organism with increased efficiency of methionine synthesis, the following definitions are given. The term "microorganism synthesis efficiency" describes the carbon yield of methionine. This efficiency is calculated as a percentage of the energy input that entered the system in the form of a carbon substrate. Throughout the invention this value is given in percentage values ((mole of methionine) (mole of carbon substrate) "1 x 100" unless otherwise indicated.) The term "increased efficiency of methionine synthesis" It is related to a comparison between an organism that has been modeled theoretically by the methods mentioned in the above and that has a higher efficiency of methionine synthesis compared to the initial model organism that was used to assign parameters. Methionine synthesis "can also describe the situation in which an organism that has been genetically modified for example provides an increased efficiency of methionine synthesis compared to the respective starting organism.The term" metabolic pathway "is related to a series of reactions that are part of the metabolic network that is used in the theoretical model mentioned in the above to design an organis or with improved methionine synthesis. The term "metabolic pathway" also describes a series of reactions that take place in a real organism. A metabolic pathway can comprise a well-known series of reactions since these are known from standard textbooks such as for example respiratory chain, glycosylation, tricarboxylic acid cycle, etc. Alternatively, the metabolic pathways may be defined separately for the purposes of the present invention. The term "metabolic flux" describes the amount of energy input that is fed into the system, for example in the form of a carbon source such as glucose and that passes through the reactions of the metabolic network of an organism or model theoretical mentioned in the above. Each reaction of the network will usually contribute to the overall metabolic flow. As a consequence, a metabolic flux can be assigned to each reaction of the network. As the elementary flow modes are calculated on the basis of the stoichiometry of the various reactions of the network model, the flows are typically given as relative molar values, normalized to the energy uptake rate which is measured in the form of glucose, say, the flows are given in mol (substance) x (mol of glucose) "1 x 100. The term" modified metabolic flux "is related to a situation in which the metabolic flux through a certain reaction or a metabolic pathway of an organism that has been genetically modified, it increases or decreases compared to the starting organism. This term is also related to the situation where, according to the theoretical method mentioned above to determine or design an optimized organism for methionine synthesis, the theoretical metabolic flux through a certain metabolic reaction or metabolic pathway increases or decreases when changing the parameters of the theoretical metabolic network. If the term "approaching the metabolic flow" is used in the context of the present invention, this is related to genetically modifying organisms in order to increase and / or decrease and / or introduce the metabolic flux through the synthesis pathways. of methionine that have been used to build the theoretical model mentioned in the above. As the genetic modifications are selected on the basis of the predictions of the model mentioned in the above, the metabolic flux of the genetically modified organisms, in comparison with the respective starting organism, must more closely resemble the metabolic flow of the optimized model mentioned above. previous. The terms "express", "expressing", "expressed" and "expression" refer to the expression of a gene product (for example, a biosynthetic enzyme of a one-way gene or reaction defined and described in this application) to a level at which the enzymatic activity resulting from this encoded protein or the pathway or reaction to which it refers allows metabolic flow through this pathway or reaction in the organism in which this gene / pathway is expressed. The expression can be made by genetic alteration of the microorganism that is used as a starting organism. In some embodiments, a microorganism can be genetically altered (e.g., engineered) to express a gene product at an increased level relative to that produced by the starting microorganism or in a comparable microorganism that has not been altered. Genetic alteration includes, but is not limited to, altering or modifying regulatory sites or sequences associated with the expression of a particular gene (for example by adding strong promoters, inducible promoters or multiple promoters or by removing regulatory sequences so that the expression is constitutive), modify the chromosomal location of a particular gene, alter nucleic acid sequences adjacent to a particular gene such as a ribosome binding site or transcription terminator, increase the copy number of a particular gene, modify proteins (for example, regulatory proteins, suppressors, enhancers, transcriptional activators and the like) involved in the transcription of a particular gene and / or translation of a particular gene product, or any other conventional means to destabilize in its regulation the expression of a particular gene using a routine in the art (including but not limited to The use of antisense nucleic acid molecules, for example, to block the expression of repressor proteins). The terms "overexpress", "overexpressing", "overexpressed" and "overexpression" refer to expression of a gene product (e.g., a methionine biosynthetic enzyme or sulfate reduction pathway enzyme or cysteine biosynthetic enzyme or a gene). or a defined route or reaction and described in this application) at a higher level than that present prior to a genetic alteration of the starting microorganism. In some embodiments, a microorganism can be genetically altered (e.g., engineered) to express a gene product at an increased level relative to that produced by the starting microorganism. Genetic alteration includes, but is not limited to, altering or modifying sequences or regulatory sites associated with the expression of a particular gene (for example, by adding strong promoters, inducible promoters or multiple promoters or by removing regulatory sequences so that the expression is constitutive), modify the chromosomal location of a particular gene, alter nucleic acid sequences adjacent to a particular gene such as a ribosome binding site or transcription terminator, increase the copy number of a particular gene, modify proteins (e.g. regulatory proteins, suppressors, enhancers, transcriptional activators and the like) involved in the transcription of a particular gene and / or translation of a particular gene product, or any other conventional means to destabilize in its regulation the expression of a particular gene using a routine in the technique (including but not limited to the use of antisense nucleic acid molecules, for example, to block the expression of repressor proteins). Examples for overexpression of genes in organisms such as C gl utami cum can be found in Eikmanns et al (Gene. (1991) 102, 93-8). In some embodiments, a microorganism may be physically or environmentally altered to express a gene product at an increased or lower level relative to the level of expression of the gene product by the starting microorganism. For example, a microorganism can be treated with or cultured in the presence of a known or suspected agent to increase the transcription of a particular gene and / or translation of a particular gene product so that the transcription and / or translation is enhanced or increased. Alternatively, a microorganism may be cultured at a selected temperature to increase the transcription of a particular gene and / or translation of a particular gene product so that the transcription and / or translation is enhanced or increased. The terms "destabilize in its regulation," "destabilized in its regulation" and "destabilization of regulation" refer to alteration or modification of at least one gene in a microorganism, where the alteration or modification results in increasing efficiency of the production of methionine in the microorganism in relation to the production of methionine in the absence of alteration or modification. In some embodiments, a gene that is altered or modified encodes an enzyme in a biosynthetic pathway, so that the level or activity of the biosynthetic enzyme in the microorganism is altered or modified. In some embodiments, at least one gene encoding an enzyme in a biosynthetic pathway is altered or modified so that the level or activity of the enzyme is enhanced or increased relative to the level in the presence of the unaltered or wild-type gene. In some modalities, the biosynthetic pathway is the methionine biosynthetic pathway. In other modalities, the biosynthetic pathway is the cysteine biosynthetic pathway. The destabilization of the regulation also includes altering the coding region of one or more genes to yield, for example, an enzyme that is resistant to feedback or has a specific higher or lower activity. Also, destabilization of the regulation also covers genetic alteration of genes that encode transcriptional factors (for example, activators, repressors) that regulate the expression of genes in the biosynthetic pathway of methionine and / or cysteine. The phrase "pathway or reaction destabilized in its regulation" refers to a pathway or biosynthetic reaction in which at least one gene encoding an enzyme in a pathway or biosynthetic reaction is altered or modified so that the level or activity of at least one biosynthetic enzyme is altered or modified. The phrase "via destabilized in its regulation" includes a biosynthetic pathway in which more than one gene has been altered or modified, thereby altering the level and / or activity of the corresponding gene products / enzymes. In some cases the ability to "destabilize in its regulation" one way (for example, to destabilize in its regulation simultaneously more than one gene in a given biosynthetic pathway) in a microorganism arises from the particular phenomenon of microorganisms in which more than one enzyme (for example, two or three biosynthetic enzymes) are encoded by genes that occur adjacent to each other in a contiguous piece of genetic material called an "operon." In other cases, in order to destabilize a pathway in their regulation, a number of genes must be destabilized in regulation in a series of sequential design stages. The term "operon" refers to a coordinated unit of genetic material that contains a promoter and possibly a regulatory element associated with one or more, preferably at least two, structural genes (e.g., genes encoding enzymes, e.g., enzymes biosynthetics). The expression of structural genes can be regulated in a coordinated manner, for example, by regulatory proteins that bind to the regulatory element or by antitermination of transcription. Structural genes can be transcribed to give a single mRNA that encodes all of the structural proteins. Due to the coordinated regulation of the genes included in an operon, the alteration or modification of the single promoter and / or regulatory element may result in alteration or modification of each gene product encoded by the operon. Alteration or modification of a regulatory element includes, but is not limited to, removing the promoter (s) and / or endogenous regulatory elements, adding strong promoters, inducible promoters or multiple promoters or removing regulatory sequences so that the expression of gene products is modify, modify the chromosomal location of the operon, alter nucleic acid sequences adjacent to the operon or within the operon such as a ribosome binding site, use of codons, increase the number of copies of the operon, modify proteins (e.g., regulatory proteins , suppressors, enhancers, transcriptional activators and the like) involved in the transcription of the operon and / or translation of the gene products of the operon, or any other conventional means to destabilize in their regulation the expression of genes routinely in the art (including, but not limited to, the use of antisense nucleic acid molecules, by example, to block the expression of repressor proteins). In some embodiments, the recombinant microorganisms described herein have been engineered to overexpress a gene or gene product derived by bacteria. The terms "derived by bacteria" and "derived from bacteria" refer to a gene that is found naturally in bacteria or a gene product that is encoded by a bacterial gene. The term "organism" for the purposes of the present invention refers to any organism that is commonly used for the production of amino acsuch as methionine. In particular, the term "organism" is related to prokaryotes, lower eukaryotes and plants. A preferred group of the organisms mentioned in the foregoing comprises actinobacteria, cyanobacteria, proteobacteria, Chloroflexus a uran tiacus, Pirellula sp. 1, halobacteria and / or methanococci, preferably corynebacteria, mycobacteria, streptomyces, salmonella, Escheri chia coli, Shigella and / or Pseudomonas. Particularly preferred microorganisms are selected from Coryneba cteri um glummicum, Escherichia coli, microorganisms of the genus Ba cillus, particularly Baci lus subtilis, and microorganisms of the genus Streptomyces. The term "initial organism" is used to describe the organism and the metabolic network that has been used to assign the initial set of parameters for the model mentioned in the foregoing according to independent claim 1. The term "starting organism" refers to the organism that is used for genetic modification to increase the confidence of methionine production. A starting organism can be either a wild-type organism or an organism that already carries mutations. The starting organism can be identical to the initial organism. The starting organisms, for example, can be methionine overproducers. The term "wild-type organism" is related to an organism that has not been genetically modified. The term overproducer of methionine is related to an organism that has been altered by genetic manipulation, by mutation and selection or by any other method known and that overproduces more methionine than the wild-type strain that was used to obtain a methionine overproducer. The organisms of the present invention, however, may also comprise yeasts such as Schizosaccharomyces pombe or cerevisiae and Pichia pastoris. Plants are also considered by the present invention for the production of amino acids. Such plants can be monocots or dicots such as crop plants, food plants or monocotyledonous or dicotyledonous forage plants. Examples for monocotyledons plants are plants that belong to the genera of oats (oats), triticum (wheat), sécale (rye), hordeum (barley), oryza (rice), panicum, pennisetum, setaria, sorghum (millet), zea (corn) and similar. Dicotyledonous cultivation plants include, among other cotton, legumes as legumes and in particular alfalfa, soybeans, rapeseed, tomato, sugar beet, potato, ornamental plants as well as trees. Additional crop plants may comprise fruits (in particular apples, pears, cherries, grapes, citrus, pineapple and bananas), oil palms, tea bushes, cocoa trees and coffee trees, tobacco, henequen as well as, relative to plants medicinal, rauwolfia and digitalis. Particularly preferred are grains of wheat, rye, oats, barley, rice, corn and millet, sugar beet, rapeseed, soy, tomato, potato and tobacco. Additional culture plants can be taken from US 6, 137, 030. The term "metabolite" refers to chemical compounds that are used in the metabolic pathways of organisms as precursors, intermediates and / or end products. Such metabolites can not only serve as chemical building blocks, but can also exert a regulatory activity on enzymes and their catalytic activity. It is known from the literature that such metabolites can inhibit or stimulate the activity of enzymes (Stryer, Biochemistry, (1995) W. H. Freeman &Company, New York, New York). For the purposes of the present invention, the term "external metabolite" comprises substrates such as glucose, sulfate, thiosulfate, sulfite, sulfur, ammonia, phosphate, metal ions such as Fe2 + Mn2 + Mg2 +, Co2 + Mo02 + and oxygen etc. In certain embodiments, the (external) metabolites comprise the so-called Cl metabolites. These latter metabolites can function, for example, as methyl donors and comprise compounds such as formate, methanol, formaldehyde, methanethiol, dimethyldisulfide etc. The term "products" comprises methionine, cysteine, glycine, lysine, trehalose, biomass, C0, etc. Before describing the invention with respect to its particular embodiments, a general introduction is given as to how the forecasts were obtained by elementary flow analysis. The elementary flow analysis begins with the formulation and implementation of all metabolic reactions relevant to growth and production of methionine. The required information can be collected from public databases such as KEGG (http://www.genome.jp/kegg/) and others. The model is then configured accordingly and reflects the natural potential of the wild type organism and serves as the starting point for further development of methionine overproductive model strains. To obtain an initial model, models of biochemical reactions for methionine synthesis were constructed. For this purpose, models were constructed that include all the relevant routes of the central carbon and sulfur metabolism that involve all relevant pathways linked to methionine production as they are known from the literature. If a pathway for a certain organism, such as for example E. coli, it is known that it does not occur in another organism such as C. gl utamicum, the reactions of specific pathways of the organism were only considered in the model for that specific organism and were omitted for the other organisms when the model for the initial organism is constructed. After an initial model has been obtained, the pathways of other organisms that are known not to occur in the model organism can then be considered, that is, introduced, for further optimization. The different biochemical reactions that contribute to a metabolic network can be obtained for example from standard textbooks, scientific literature or Internet links such as http: // www. genome jp / kegg / metabolism. html An analysis in elementary flow mode was then performed as described in the literature (see for example Papin et al. (2004) vide supra, Schilling et al. (2000) vide supra, Schuster et al. (1999) vide supra). The modes of elementary flow are calculated on the basis of the stoichiometry of the various reactions. The specific kinetics of each reaction is usually not taken into consideration. When constructed, a metabolic network typically comprises endless cycles of reversible pathways and reactions. Various routes of the tracks can be taken in this way in order to arrive at a compound such as methionine. In this way, depending on which route is taken, the energy requirements for production of the same compound can change within the same network. As a consequence, if the various reactions of a network are described by parameters and put into an algorithm such as METATOOL software (Pfeiffer et al. (1999) Bioinforma ti cs, 153, 251-257; Schuster et al. (1999) vide supra), the network can be modified and optimized in order to identify the route that allows the most efficient methionine synthesis. For the purposes of the present invention, the analysis of metabolic pathways was carried out using the METATOOL program. The version used for the present invention (meta 4.0.l_double.ex) is available on the Internet at http: //www.biozentrum.uni-wuerzburg. from / bioinformatik / computing / metatool / pinguin. biology uni-j ena from / bioinformatik / networks /. The mathematical details of the algorithm are described by Pfeiffer et al.

(Pfeiffer et al. (1999) vide supra). If the analysis of metabolic pathways is carried out using the METATOOL program, several hundred modes of elementary flow result for each situation investigated. For each of these flow modes, the carbon yields of methionine, as indicated in the above, were calculated as a percentage of the carbon that entered the system as a substrate. For the various flow modes, the carbon yield of the biomass can be calculated as a percentage of the energy that entered the system as a carbon substrate. This parameter can be calculated in this way as ((mole of biomass) (mole of substrate) -1 x 100). Co-substrates other than glucose, such as formate, formaldehyde, methanol, methanethiol or its dimethyldisulfide dimer can also be considered accordingly. The comparative analysis of all these modes of elementary flow obtained for a certain network scenario then allows the determination of the maximum theoretical efficiency for the synthesis of methionine. In this way, one obtains a theoretical prediction of the optimal metabolic flow through the metabolic network of an organism that must have an optimal efficiency for the synthesis of methionine. The details of such theoretical analysis of metabolic flux are described in the experimental section. The method for determining or theoretically designing such an organism with increased efficiency for methionine synthesis constitutes the subject of independent claim 1. The theoretical forecasts obtained by these methods can then be put into practice by genetically modifying the respective organism in order to intensify or reduce the metabolic flux through those routes identified by the forecast model. Surprisingly, theoretical predictions can also be put into practice according to model predictions by genetically altering a starting organism, which is not identical to the initial organism. Such a starting organism in this way may not be a wild-type organism, but organisms that are already genetically modified. In one embodiment, the starting organism may be, for example, a methionine overproducer, ie, a genetically modified organism that is already known to produce more methionine than the respective wild-type organism. Although theoretical predictions have not been calculated for such a methionine overproducer, they still allow building genetically modified organisms on the basis of the methionine overproducer that provides an increased efficiency of methionine synthesis. If, for example, theoretical predictions imply that methionine synthesis is more efficient if the metabolic flux through the pentose phosphate pathway (PPP) is increased, an organism is genetically modified for that purpose. This can be done, for example by increasing the amount and / or activity of the enzymes that catalyze certain stages of the PPP in order to channel more metabolic flow through this path compared to a non-genetically modified organism that is grown from another way exactly under the same conditions. The flow to the PPP can also be intensified, for example, by downregulating the enzymatic activity in an irreversible reaction of another parallel pathway that redirects the metabolic flow to the PPP. The flow through the PPP can also be intensified by introducing specific mutations in genes that encode proteins that are involved in enzymes of the PPP cycle such as mutations in pyruvate carboxylase as described by Onishi et al. (Appl Mi crobiol Biotechnol. (2002), 58, 217-23). These altered genes contain mutations compared to genes derived from so-called wild-type strains. These mutations can lead to altered enzymatic activity or sensitivity to molecular feedback inhibitors. Correspondingly, if the theoretical model requires a reduction of the metabolic flow to the pentose phosphate pathway, the quantity and / or activity of the enzymes of this pathway can be reduced. Metabolic flux analysis can also be used to transfer the results generated from one organism to another. In this way, if it is found by analysis in elementary flow mode, for example in E. A certain pathway with increased activity is crucial for an efficient synthesis of methionine, and if this pathway is obviously not used or does not occur in another organism such as C. glutamicum, this pathway can be introduced into the respective organism by introducing the genes that encode the enzymatic activities of this route in the respective organism. By this methodology, it is not only possible to optimize the microorganisms with respect to the synthesis of methionine by optimizing their endogenous metabolic pathways, but also to introduce an exogenous metabolic pathway in order to further intensify the synthesis of methionine and / or increase the efficiency in synthesis. In view of this situation, the present invention also relates to a method for producing an organism that is selected from the group of prokaryotes, lower eukaryotes and plants with increased efficiency of methionine synthesis compared to the starting organism comprising the steps of : to. carry out the analysis in elementary flow mode mentioned above to obtain a theoretical forecast about the optimal metabolic flux distribution in an organism that is optimized with respect to the methionine synthesis, and b. genetically modify an organism in a determined way to modify the metabolic pathways existing in the organism so that the metabolic flow of the organism approximates the theoretical model of stage a) in comparison with the organism of initiation and / or c. genetically modify an organism in a determined way to introduce exogenous metabolic pathways in the organism so that the metabolic flux of the organism approximates the theoretical model of stage a) in comparison with the organism of initiation and / or d. provide external metabolites in sufficient quantities to channel the metabolic flow through the metabolic pathways of b) and e). A further aspect of the present invention relates to a method that implements the theoretical predictions of flow distribution in an organism that is optimized for methionine synthesis by producing an organism that is selected from the group of prokaryotes, lower eukaryotes, and plants at : • modify metabolic flux through at least one of the following metabolic pathways by genetic modification of organisms: • phosphotransferase system (PTS) and / or pentose phosphate pathway (PPP) and / or glycolysis (EMP ) and / or tricarboxylic acid cycle (TCA) and / or glyoxylate deviation (GS) and / or • anaplerosis (AP) and / or respiratory chain (RC) and / or sulfur assimilation (SA) and / or synthesis of methionine (MS) and / or serine / cysteine / glycine synthesis (SCGS) and / or • glycine cleavage system (GCS) and / or • transhydrogenase conversion (THGC) and / or • via 1 (Pl) and / or • track 2 (P2) and / or • track 3 (P3) and / or • track 4 (P4) and / or • track 5 (P5) and / or • track 6 (P6) and / or • track 7 (P7), and / or • enter a metabolic flow through at least one of the following exogenous metabolic pathways by genetic modification of organisms: • glycine cleavage system (GCS) and / or • transhydrogenase conversion (THGC) and / or • thiosulfate reductase system (TRS) and / or • sulfate reductase system ( SARS) and / or • sulfite reductase system (SRS) and / or • formate converter system (FCS) and / or • methanethiol converter system (MCS), and / or • culture organisms in the presence of: a. sulfate and / or b. sulfite and / or c. sulfur and / or d. thiosulfate and / or e. compounds containing organic sulfur and / or f. Cl metabolites such as formate, formaldehyde, methanol, methanethiol or its dimethyl disulfide dimer. Organisms that have been genetically modified for the purpose of implementing the prognoses for a model organism with increased efficiency of methionine biosynthesis are also an object of the present invention. As mentioned in the above, for the calculation of the optimal metabolic flow through a metabolic network for methionine synthesis, the specific metabolic pathways of the organism that lead to this amino acid are used. Additionally, specific stoichiometries of specific organisms must be considered for each constructed metabolic network. The stoichiometries can be taken from the sources mentioned in the above. Even when such metabolic networks can differ between organisms such as E. coli to C. glutamicum, Figure 1 shows a set of reactions that were used to calculate the initial metabolic network using C. gl utami cum as an example. As this set is only a minimum set of reactions for a metabolic network that contributes to the synthesis of methionine, additional pathways were considered for other organisms such as E. coli if their existence was known. However, if the following generally refers to a certain metabolic pathway or a specific enzyme, these general references are all related to the reactions shown in Figure 1 unless otherwise indicated. This seems justified since these reactions were largely identical in E. coli and C. glutamicum. For the purposes of the present invention, the various reactions are grouped into the following groups of routes: phosphotransferase system (PTS) via pentose phosphate (PPP) glycolysis (EMP) • tricarboxylic acid cycle (TCA) glyoxylate deviation (GS) anaplerosis (AP) respiratory chain (RC) sulfur assimilation (SA) • synthesis of methionine (MS) synthesis of serine / cysteine / glycine (SCGS) glycine cleavage system (GCS) conversion of transhydrogenase (THGC) via 1 (Pl) • lane 2 (P2) lane 3 (P3) lane 4 (P4) lane 5 (P5) lane 6 (P6) • lane 7 (P7) • thiosulfate reductase system (TRS) • sulfate reductase system ( SARS) • sulfite reductase system (SRS) • formate converter system (FCS) • methanethiol converter system (MCS). The simple pathways can be subdivided into the following reactions that are catalyzed by enzymes designated Rn. Abbreviations are used to define these reactions. The manner in which these definitions are to be understood for the purposes of the invention is explained with respect to the phosphotransferase system. This explanation also applies to the other reactions. For the purposes of the present invention, the phosphotransferase system (PTS) comprises the reaction of external glucose to glucose-6-phosphate (G6P). This reaction is catalyzed by the Rl enzyme which is phosphotransferase. This enzyme uses phosphoenolpyruvate as a donor of phosphorus groups (see Figure 1). For the purposes of the invention this reaction is described as: Rl in order to produce more G6P The individual reactions of the various pathways mentioned in the above in this manner are defined with respect to the enzymes that catalyze the reaction and the products resulting from the reactions. It is not indicated whether such reaction may or may not require energy in the form of ATP, NADH and / or NADPH or other co-factors, but can be taken from Figure 1. Specific stoichiometry is also not indicated, as this may vary from organism to organism. In general, the educts and energy input of the reaction are also not indicated. These data can be taken from standard textbooks or scientific publications about the various organisms. For the purposes of the present invention, the pentose phosphate pathway is characterized by the following reactions: R3 in order to produce GLC-LAC R4 in order to produce 6-P-Gluconate R5 in order to produce RIB-5P R6 in order to produce XYL-5P R7 in order to produce RIBO-5P R8 in order to produce S7P and GA3P R9 in order to produce E-4p and F6P RIO in order to produce F6P and GA3P R2 with the In order to produce G6P For the purposes of the present invention, the pathway of glycolysis (EMP) is characterized by the following reactions: Rll in order to produce F-1,6-BP R13 in order to produce DHAP and GA3P R14 in order to produce GA3P R15 in order to produce 1,3-PG R16 in order to produce 3-PG R17 in order to produce 2-PG R18 in order to produce R19 PEP in order to produce Pyr For For the purposes of the present invention, the tricarboxylic acid (TCA) cycle is defined by the following reactions: R20 with the In order to produce Ac-CoA R21 in order to produce CIT R22 in order to produce Cis-ACO R23 in order to produce ICI R24 in order to produce 2-OXO R26 in order to produce SUCC-CoA R27 with the In order to produce SUCC R28 in order to produce FUM R29 in order to produce MAL R30 in order to produce OAA For the purposes of the present invention, the glyoxylate (GS) deviation pathway is defined by the following reactions: R21 in order to produce CIT R22 in order to produce Cis-ACO R23 in order to produce ICI R31 in order to produce GLYOXY and SUCC R32 in order to produce MAL R28 in order to produce FUM R29 in order to produce produce MAL R30 in order to produce OAA For the purposes of the present invention, the path of anaplerosis (AP) is defined by the following reactions: R34 in order to produce OAA R33 / R36 in order to produce OAA for the purposes of the present invention, the respiratory chain (RC) is defined by the following reactions: R59 that catalyzes: 2NADH + 02ex + 4ADP = 2NAD + 4ATP R60 that catalyzes: 2FADH + 02ex + 2 ADP = 2FAD + 2ATP For the purposes of the present invention, the sulfur assimilation route (SA) is defined by the following reactions: R55 in order to produce H2SO3 R58 in order to produce H2S For the purposes of the present invention, the synthesis route Methionine (MS) is defined by the following reactions: R37 in order to produce Asp R47 in order to produce ASP-P R48 in order to produce ASP-SA R39 in order to produce HOM R40 in order to produce O-AC-HOM R46 in order to produce CYSTA R49 in order to produce HMOCYS R54 in order to produce HOMOCYS R52 in order to produce MET R53 in order to produce METex. For the purposes of the present invention, the of serine / cysteine / glycine synthesis (SCGS) is defined by the following reactions: R41 in order to produce 3-PHP R42 in order to produce SER-P R43 in order to produce SER R44 in order to produce OR -AC-SER R45 in order to produce CYS R38 in order to produce ucir M-THF and Glicinaex For the purposes of the present invention, pathway 1 (Pl) comprises the following reactions: R25 in order to produce GLU For the purposes of the present invention, lane 2 (P2) comprises the following reactions: R33 / R36 in order to produce OAA R30 in order to produce MAL R57 in order to produce PYR + C02 For the purposes of the present invention, lane 3 (P3) ) comprises the following reaction: R56 which catalyzes: ATP = ADP For purposes of the present invention, line 4 (P4) comprises the following reactions: R62 which catalyzes: GTP + ADP = ATP + GDP For the purposes of the present invention , lane 5 (P5) is defined by the following reactions: R50 catalyzing: ATP + acetate = ADP + acetyl For the purposes of the present invention, lane 6 (P6) comprises the following reactions: R51 catalyzing: acetyl P + HCoA = Acetyl CoA For purposes of the present invention, lane 7 (P7) comprises the following reaction: R61 which catalyzes: 6231 NH3ex + 233 S04ex + 205 G6P + 308 F6P + 879 RIB0-5P + 268 E4P + 129 GA3P + 1295 3-PG + 652 PEP + 2604 PYR + 3177 AC-CoA + 1680 OAA + 1224 2-OXO + 16429 NADPH = BIOMASSex + 16429 NADP + 3177 H-CoA + 1227 C02e? As stipulated in the above, the stoichiometry will vary from organism to organism and can be taken from the literature or the Internet pages mentioned in the above. Additionally, the metabolic network of certain organisms such as E. coli or C. gl utami cum may comprise additional reaction pathways. Such additional routes, as used for the purposes of the present invention, include: • glycine cleavage system (GCS) and / or • transhydrogenase conversion (THGC) and / or • thiosulfate reductase system (TRS) and / or • sulfate reductase system (SARS) and / or • sulfite reductase system (SRS) and / or • formate converter system (FCS) and / or • methanethiol converter system (MCS) For the purposes of the present invention, the glycine cleavage system (GCS) comprises the following reactions: R71: in order to produce M-HPL R72: in order to produce Methylene-THF The person skilled in the art knows very well that the reactions of R71 and R72 are catalyze at least three proteins, ie gcvH, P and T (see Tables 1 and 2) . gcvP catalyzes the decarboxylation of glycine to C02 and an aminomethyl group, while GcvH is a carrier of the aminomethyl group (R71). A description of the glycine cleavage system can be found in Neidhardt F.C. (1996) E. coli and S. typhimuri um, ASM Press Washington. gcvT is involved in the transfer of the Cl unit from protein H to tetrahydrofolate and the release of NH3 (R72).

The reaction is then typically completed by the fourth subunit which is lipoamide dehydrogenase. The lipoamide dehydrogenase encoded by lpdA functions as the electronic transfer of NAD to NADH. This dehydrogenase is borrowed from the multiple subunit pyruvate dehydrogenase and is commonly called lpdA. For the purposes of the present invention the GCS in this manner can be summarized as: R71 / 72: in order to produce Methylene-THF For the purposes of the present invention, the GCS can optionally also comprise the following additional reaction: R78: with in order to produce Methyl-THF Strictly speaking, R78 does not belong to GCS since it only serves to provide Methyl-THF. However, in organisms in which R78 does not occur, R78 can be implemented along with the other GCS reactions. In organisms in which R78 already occurs, this may not be necessary. For purposes of the present invention, the transhydrogenase conversion system (THGC) comprises the following reaction: R70: in order to produce NADPH For the purposes of the present invention, THGC may also comprise the following reaction: R81: with in order to produce NADPH While R70 for example can be a cytosolic transhydrogenase, R81 for example can be a transmembrane transhydrogenase. For the purposes of the present invention, the thiosulfate reductase system (TRS) comprises the following reactions: R73: in order to metabolize thiosulfate to sulfide and sulfite For the purposes of the present invention, the TRS may additionally comprise: R82: with in order to import extracellular H2S2O3 into the cell and / or R45a: in order to produce more S-Sulfocysteine and / or R49: in order to produce more S-Sulfocysteine. R45a and / or R49 convert Thiosulfate to S-Sulfo-Cysteine and thus belong to the SRS. For the purposes of the present invention, the sulfate reductase (SARS) system comprises the following reaction: R80: in order to metabolize sulfate to sulfite For the purposes of the present invention, the sulfite reductase (SRS) system comprises the following reaction: R74 in order to metabolize sulfite to sulfide For the purposes of the present invention, the formate converting system (FCS) comprises the following reactions: R75: in order to produce 10-formyl-THF R76: in order to produce Methylene-THF R78: in order to produce Methyl-THF For the purposes of the present invention, the methanethiol converting system (MCS) comprises the following reactions: R77 for the purpose of methyl sulfhydryl O-Acetyl-homoserine with methanethiol For the purposes of the present invention, the 8 (P8) pathway comprises the following reaction: R79 with in order to degrade formyl-THF to formate and tetrahydrofolate Specific examples for the enzymes mentioned in the above are given in the following table. Additional reactions can be found in the introduction of reactions given further in the following.

Table 1 The access numbers in the above are the official access numbers of the Genbank or are synonyms for access numbers that are cross-referenced in the Genbank. These numbers can be found and found at http://www.ncbi.nlm.nih.gov/. The present invention also envisions that the metabolic flux through other pathways and reactions can be modulated by theoretical or genetic manipulation of organisms to produce organisms with increased efficiency of methionine synthesis as long as it is known, for example in the scientific literature, that these reactions participate directly or indirectly in the methionine synthesis. These pathways and reactions, of course, can also be implemented in the theoretical analysis in elementary flow mode. Organisms (genetically modified) obtained by these methods are also part of the invention. As mentioned in the foregoing, in accordance with the present invention the actual metabolic flux in an organism must be approximated to the optimum theoretical flux for an organism with increased methionine synthesis, as determined by the analysis in the elemental flux mode of According to claim 1. For purposes of the present invention, "approximate" means that the metabolic flux of the genetically modified organism as a result of the genetic modification is more similar to the metabolic flux of the theoretical predictions than the metabolic flux does. of the starting organism. As already stipulated, the modulation of the metabolic flux of the starting organism can be influenced by the genetic alteration of the organism, for example by influencing the quantity and / or the activity of the enzymes that catalyze specific reactions of the considered network. Additionally, the metabolic flux can be influenced by the use of certain nutrients and external metabolites such as sulfate, thiosulfate, sulfite and sulfur and Cl compounds such as formate-formaldehyde, methanol-methanethiol and dimethyldisulfide. While the influence of external metabolites such as thiosulfate, formate or methanethiol will be explained in more detail later, general examples are given in the following for the genetic modification of organisms. In the following, it will be explained with respect to a specific reaction how the metabolic flux through a certain way can be channeled by the genetic modification of an organism. These explanations apply correspondingly to other reactions. If, for example, the theoretical model obtained or the model organism designed according to the method of the present invention predicts that for an efficient synthesis of methionine the metabolic flux should mainly be channeled towards the PPP, a real organism with increased metabolic flux through This can be obtained by genetically influencing the amount and / or activity of the reactions mentioned above that are part of the PPP. In this way, the metabolic flux can increase towards PPP by increasing the amount and / or activity of R3, which leads to the formation of more GLC-LAC. In the same way, increasing the amount and / or activity of R4, R5, R6, R7, R8, R9 or RIO can increase the metabolic flux towards PPP. The same can be achieved by increasing the activity of R2 towards the production of G6P. If the theoretical model obtained by the method of the present invention predicts a reduction of the metabolic flux through the TCA, this can be achieved by reducing the amount and / or activity of the following enzymes R21, R22, R23, R24, R26, R27, R28, R29 or R30. How it is that the activity and / or quantity of an enzyme can be increased or reduced is apparent to the experienced person and will also be exemplified in the following. For general purposes, however, it should be noted that in a metabolic pathway, as in Figure 1, certain reactions can be considered as irreversible. While almost any reaction of a biological network is an equilibrium reaction that is able to proceed in both directions, irreversible reactions are commonly considered as reactions in which, by the contribution of for example energy, the reaction is predominantly oriented in one direction, so that the equilibrium of the reaction lies almost exclusively on one side of the reaction. In the case of PPP, such irreversible reactions, for example, are reactions catalyzed by R3 and R5, both of which are favored by the formation of NADPH. Other irreversible reactions, since this term is used in the context of this invention, for example are R16 of the EMP, R24 of the TCA, etc. Irreversible reactions are indicated in Figure 1 by arrows pointing in only one direction. If, in the context of the present invention, it is stated that the metabolic flux through a certain reaction pathway is increased by increasing the amount and / or activity of the catalyzing enzyme in that direction, then this assertion must be seen in the context of how reactions are defined in the above. Increasing or decreasing the amount and / or activity of an enzyme must be understood with respect to the direction in which the reaction must also be pushed or channeled. Since the reactions of the various pathways of the metabolic network according to the present invention have been defined by an enzyme and the product that is formed by that enzyme, it is clearly understood by the person skilled in the art to increase the amount and / or activity of an enzyme or decrease the amount and / or activity of an enzyme, influences the amount and / or activity of the enzyme in such a way that more or less product is obtained. In this way, if for example it is stated that the activity of the R6 enzyme increases, then in view of the description of this reaction mentioned in the above, this means that by increasing the amount and / or activity of R6, the amount of XYL -5P is increased. Similarly, if it is stated that the amount and / or activity of R23 is increased, this refers to a situation where the amount and / or activity of R23 is increased to produce more ICI. Correspondingly, if for example the amount and / or activity of R29 decreases, then this means that the amount and / or activity of R29 is reduced in order to produce less MAL. If the theoretical model organisms with increased efficiency in methionine require for example an increase in metabolic flux through a certain pathway, in one embodiment of the invention it may be sufficient to modify the amount and / or activity of only one enzyme in that reaction pathway. Alternatively, the amount and / or activity of various enzymes of this metabolic pathway can be modified. If, for example, the theoretical model obtained by elementary flow analysis suggests for example increasing the metabolic flux through the PPP and the TCA while the metabolic flux through the CR should be reduced, this can be achieved by increasing the quantity and / or activity of only one enzyme of the PPP and the TCA cycle while the activity and / or amount of only one enzyme of the RC can be reduced. Alternatively, the amount and / or activity of various or all enzymes of these routes can be influenced at the same time. The person skilled in the art will also realize that what can be defined in the foregoing as an enzymatic reaction that is carried out by a single enzymatic activity, can in fact be a series of enzymatic (sub) steps by various enzymes that they all provide the indicated global activity (for example sulphite or thiosulfate reductase). The overall enzymatic activity indicated (see the R numbers in the above) can also be composed of various subunits. In this case the metabolic flux through the reactions identified in the above can be influenced by modifying the activity and / or amount of at least one of the enzymes that carry out one of the only (sub) stages or at least one of the subunits. Therefore, the genes encoding (sub) stages or subunits can be considered as part of the overall respective enzymatic activity. With respect to the glycine cleavage system, the experienced person knows that the genes gcvT, and / or H, and / or P and / or L (lpdA) (see Tables 1 and 2) are involved in this system. The metabolic flux through this system that is defined in the above by the reactions R71, R72 and R71 / R72 in this way can be increased or introduced for example by over-expression of at least one of the genes identified in the above or their counterparts Increasing metabolic flux can also be achieved by over-expressing four of these genes or only two or three of these genes. Genes can be overexpressed together, for example in a naturally occurring operon or in an artificial operon constructed using promoters. Additionally, it may also be useful to overexpress the lpdA gene together with the genes gcvH, P, T (see Tables 1 and 2 again). With respect to the methionine synthesis system, the experienced person knows that the reactions: R47, R48, R39, R40 R46, R49, R52, R53, R54 are involved in the synthesis of methionine. For overexpression of transhydrogenase (R70 and R81) at least one of the udh, pntA and / or pntB genes or their homologs (see Tables 1 and 2) can be overexpressed. The genes can also be overexpressed together for example in a naturally occurring operon or in an artificial operon constructed using promoters. One can, of course, in addition or alternatively also overexpress a gene for a transmembrane transhydrogenase such as udhA and pntA, B. For overexpression of the Thiosulfate-Reductase (R73, R45a, R49 and / or R82) the subunit B genes of Cytochrome thiosulfate reductase, thiosulfate reductase electron transport protein and / or thiosulfate reductase precursor can be overexpressed either alone or in combination for example in a naturally occurring operon or in an artificial operon constructed using promoters. For these purposes the genes phsA, B and / or C or their homologs can be used (see Tables 1 and 2). Similarly, the genes of an ABC transporter such as YP_224929 can be overexpressed. For overexpression of the pentose phosphate pathway, the Glucose-6-phosphate dehydrogenase, OPCA, transketolase, transaldolase, 6-phosphogluconolactone dehydrogenase genes or their homologues (see Tables 1 and 2) can be overexpressed either alone or in any combination of 2, 3, 4 or more genes for example in a naturally occurring operon or in an artificial operon constructed using promoters. For overexpression of the sulfite reduction system (R74) the anaerobic sulfite reductase genes subunit A, B and C can be overexpressed either alone or together, for example in a naturally occurring operon or in an artificial operon constructed using promoters. The genes dsrA, dsrB and / or dsrC or their homologs (see Tables 1 and 2) can be used for these purposes. With respect to the formate converting system (FCS), the metabolic flux can be modified and in some modalities increased or introduced by modifying the amount and / or activity of at least one of the following genes that are selected from the group of Formiato-THF- ligase, Formyl-THF-cycloligase, Methylene-THF-dehydrogenase, 5,10-Methylene-THF-reductase, Methylene-THF-Reductase. Homologs thereof can also be used (see Tables 1 and 2). Metabolic flux through FCS can also be increased by overexpression of any of these genes. The sulfate reductase system (SARS, R80) can be considered to consist of sulfate-adenylate transferase subunit 1 (NP_602005) and sulfate-adenylate transferase subunit 2 (NP_602006) which constitutes the ATP: adenylyltransferase sulfate, adenosine 5'-phosphosulfate kinase (EC: 2.7.1.25), 3 '-phosphoadenosine 5' -phosphosulfate (PAPS) reductase (EC: 1.8.4.8), NCgl2717) and sulfite reductase, (EC: 1.8.1.2, CAF20840) A preferred target for modification may be the amount and / or activity of enzymes that are considered to be irreversible within the meaning of the present invention. In this way, the theoretical models obtained by the analysis of metabolic flux for organisms that show an increased efficiency for methionine synthesis give the person experienced in the technique a clear guide on what genetic manipulations the experienced person should consider to obtain a microorganism. with such optimized metabolic flux. The person skilled in the art will then distinguish the decisive enzymes that are all well known to him from constructing the theoretical metabolic network and will influence the amount and / or activity of these enzymes by the genetic modification of the organism. How such organisms can be obtained by genetic modification belongs to general knowledge in the art. o By genetically corrected organisms according to the present invention, the metabolic flux in these organisms can be corrected in order to increase the efficiency of methionine synthesis so that these organisms are characterized in that methionine is produced with an efficiency of less 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50 %, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80% or at least 85%. In the theoretical part of the experimental section we describe what resembles the optimal metabolic flow modes for C. gl utamicum and E. coli with increased efficiency of methionine production. While there is stipulated in detail how these models were calculated, what reactions were considered and what stoichiometries were used, the general conclusions from these models are listed in the following. The following section should therefore be understood as an instruction for the person skilled in the art, whose metabolic pathways must be genetically modified in order to approximate the metabolic flow through these routes to the optimum values as obtained for the theoretical model . A work schedule will then be given in the practical part of the experimental section to illustrate, for certain enzymes, what specific measure should be taken for genetic manipulation.

C. glutami cum One object of the present invention relates to a microorganism of the genus Coryneba cteri um that has been genetically modified in order to increase and / or introduce a metabolic flux through at least one of the following pathways in comparison with the starting organism: • phosphotransferase system (PTS) and / or • pentose phosphate pathway (PPP) and / or • sulfur assimilation (SA) and / or anaplerosis pathway (AP) and / or via synthesis of methionine (MS) and / or serine / cysteine / glycine synthesis (SCGS) and / or glycine cleavage system (GCS) and / or transhydrogenase conversion (THGC) and / or via 1 (Pl) and / or via 2 (P2) and / or thiosulfate reductase system (TRS) and / or sulfite reductase system (SRS) and / or • sulfate reductase system (SARS) and / or formate converter system (FCS) and / or converter system of methanethiol (MCS) and / or At the same time such optimized microorganism must optionally have at least a reduced metabolic flux or through at least one of the following routes: glycolysis (EMP) and / or tricarboxylic acid cycle (TCA) and / or glyoxylate deviation (GS) and / or respiratory chain (RC) and / or • via 3 (P3) and / or via 4 (P4) and / or via 7 (P7) and / or R19 in order to produce less pyruvate and / or R35 in order to produce less PEP and / or • R79 in order to produce less THF. pathway of anaplerosis (AP) and / or via methionine synthesis (MS) and / or serine / cysteine / glycine synthesis (SCGS) and / or glycine cleavage system (GCS) and / or transhydrogenase conversion (THGC) and / or via 1 (Pl) and / or via 2 (P2) and / or thiosulfate reductase system (TRS) and / or sulfite reductase system (SRS) and / or • sulfate reductase system (SARS) and / or Formate Converting System (FCS) and / or Methanethiol Converting System (MCS) and / or At the same time such optimized microorganism must optionally have at least a reduced metabolic flux through at least one of the following routes: glycolysis ( EMP) and / or cycle of tricarboxylic acids (TCA) and / or deviation of glyoxylate (GS) and / or respiratory chain (RC) and / or • via 3 (P3) and / or via 4 (P4) and / or via 7 (P7) and / or R19 in order to produce less pyruvate and / or R35 in order to produce less PEP and / or • R79 in order to produce less THF.

The present invention relates to a method for producing a microorganism of the genus Corynebacterium with increased production efficiency of methionine comprising the following steps. • increase and / or introduce metabolic flux through at least one of the following pathways compared to the start by genetic modification of the organism: phosphotransferase system (PTS) and / or • pentose phosphate pathway (PPP) and / or assimilation of sulfur (SA) and / or pathway of anaplerosis (PA) and / or via methionine synthesis (MS) and / or serine / cysteine / glycine system (SCGS) and / or • glycine cleavage system (GCS) and / or transhydrogenase conversion (THGC) and / or pathway 1 (Pl) and / or pathway 2 (P2) and / or thiosulfate reductase system (TRS) and / or • sulfite reductase system (SRS) and / or sulfate reductase system (SARS) and / or formate converter system (FCS) and / or methanethiol converter system (MCS) and / or at least partially decrease the metabolic flux through at least one of the following pathways compared to the start by the genetic modification of the organism: glycolysis (EMP) and / or tricarboxylic acid cycle (TCA) and / or deviation of glyoxylate (GS) and / or respiratory chain (RC) and / or pathway 3 (P3) and / or pathway 4 (P4) and / or pathway 7 (P7) and / or • R19 in order to produce less pyruvate and / or R35 in order to produce less PEP and / or R79 in order to produce less THF. One embodiment of the present invention relates to a method for producing a microorganism of the genus Coryneba cterium with an increased efficiency for methionine synthesis wherein with respect to PTS, the amount and / or activity of the following enzyme is increased and / or introduced. in comparison with the starting body: a. Rl in order to produce more G6P; and / or with respect to PPP, the amount and / or activity of the following enzymes is increased / increased and / or introduced compared to the starting organism: a. R3 in order to produce more GLC-LAC and / or b. R4 in order to produce more 6-P-Gluconate and / or c. R5 in order to produce more RIB-5P and / or d. R6 in order to produce more XYL-5P and / or e. R7 in order to produce more RIB0-5P and / or f. R8 in order to produce more S7P and GA3P and / or g. R9 in order to produce more E-4p and F6P and / or h. RIO in order to produce more F6P and GA3P and / or i. R2 in order to produce more G6P; and / or with respect to SA, the amount and / or activity of the following enzymes is increased / increased and / or introduced compared to the starting organism: a. R55 in order to produce more H2S03 and / or b. R58 in order to produce more H2S and / or with respect to AP, the amount and / or activity of the following enzyme is increased and / or introduced compared to the starting organism: a. R33 in order to produce more OAA and / or with respect to MS, the amount and / or activity of the following enzyme is increased and / or introduced compared to the starting organism: a. R37 in order to produce more Asp and / or b. R39 in order to produce more HOM and / or c. R40 in order to produce more O-AC-HOM and / or d. R46 in order to produce more CYSTA and / or e. R47 in order to produce more ASP-P and / or f. R48 in order to produce more ASP-SA and / or g. R49 in order to produce more HOMOCYS and / or h. R52 in order to produce more MET and / or i. R53 in order to produce more METex and / or j. R54 in order to produce more HOMOCYS and / or with respect to SCGS, the amount and / or activity of the following enzyme is increased and / or introduced compared to the starting organism: a. R38 in order to produce more M-THF and Glycine and / or b. R44 in order to produce more O-AC-SER and / or c. R45 in order to produce more CYS and / or with respect to CGS, the amount and / or activity of the following enzymes is increased / increased and / or introduced compared to the starting organism: a. R71 in order to produce more M-HPL and / or b. R72 in order to produce more methylene-THF c. R78 in order to produce more Methylene-THF and / or with respect to THGC, the amount and / or activity of the following enzyme is increased and / or introduced compared to the starting organism: a. R70 in order to produce more NADPH and / or b. R81 in order to produce more NADPH and / or with respect to Pl, the amount and / or activity of the following enzymes is increased / increased and / or introduced compared to the starting organism: a. R25 in order to produce more Glu; and / or with respect to P2, the amount and / or activity of the following enzymes is increased / increased and / or introduced compared to the starting organism: a. R33 and / or R36 in order to produce more OAA and / or b. R30 in order to produce more MAL and / or c. R57 in order to produce more Pyr; and / or with respect to TRS, the amount and / or activity of the following enzyme is increased and / or introduced compared to the starting organism: a. R73 in order to metabolize thiosulfate to sulfur and sulphite and / or b. R82 to transport more external HS203 towards the cell and / or with respect to SRS, the amount and / or activity of the following enzyme is increased as compared to and / or introduced to the starting organism: a. R74 in order to metabolize sulfite to sulfide and / or with respect to FCS, the amount and / or activity of the following enzymes is increased / increased and / or introduced compared to the starting organism: a. R75 in order to produce 10-formyl-THF and / or b. R76 in order to produce Methylene-THF from 10-formyl-THF and / or c. R78 in order to produce more Methylene-THF and / or with respect to MCS, the amount and / or activity of the following enzyme is increased and / or introduced compared to the starting organism: a. R77 for the purpose of methyl sulfhydryl O-Acetyl-homoserine with methanethiol and / or with respect to SARS, the amount and / or activity of the following enzyme (s) is increased / increased and / or introduced compared to the starting organism : to. R80 in order to produce more sulfite and / or with respect to EMP, the amount and / or activity of the following enzyme (s) is reduced / reduced at least partially compared to the starting organism: a. Rll in order to produce less F-1,6-BP and / or b. R13 in order to produce less DHAP and GA3P and / or c. R14 in order to produce less GA3P and / or d. R15 in order to produce less 1,3-PG and / or e. R16 in order to produce less 3-PG and / or f. R17 in order to produce less 2-PG and / or g. R18 in order to produce less PEP and / or h. R19 in order to produce less Pyr; and / or with respect to TCA, the amount and / or activity of the following enzyme (s) is reduced / reduced at least partially as compared to the starting organism: a. R20 in order to produce less Ac-CoA and / or b. R21 in order to produce less CIT and / or c. R22 in order to produce less Cis-ACO and / or d. R23 in order to produce less ICI and / or e. R24 in order to produce less 2-0X0 and / or f. R26 in order to produce less SUCC-CoA and / or g. R27 in order to produce less SUCC and / or h. R28 in order to produce less FUM and / or i. R29 in order to produce less BAD and / or j. R30 in order to produce less OAA; and / or with respect to GS, the amount and / or activity of the following enzyme (s) is reduced / reduced at least partially as compared to the starting organism: a. R21 in order to produce less CIT and / or b. R22 in order to produce less Cis-ACO and / or c. R23 in order to produce less ICI and / or d. R31 in order to produce less GLYOXY and SUCC and / or e. R32 in order to produce less BAD and / or f. R28 in order to produce less FUM and / or g. R29 in order to produce less BAD and / or h. R30 in order to produce less OAA; and / or with respect to RC, the amount and / or activity of the following enzyme (s) is reduced / reduced at least partially as compared to the starting organism: a. R60; and / or the amount and / or activity of the following enzyme (s) is reduced / reduced at least partially as compared to the starting organism: a. R19; and / or the amount and / or activity of the following enzyme (s) is reduced / reduced at least partially as compared to the starting organism: a. R35; and / or with respect to P3, the amount and / or activity of the following enzyme (s) is reduced / reduced at least partially as compared to the starting organism: a. R56; and / or with respect to P4, the amount and / or activity of the following enzyme (s) is reduced / reduced at least partially as compared to the starting organism: a. R62; and / or with respect to P7, the amount and / or activity of the following enzyme (s) is reduced / reduced at least partially as compared to the starting organism: a. R61, and / or with respect to P8, the amount and / or activity of the following enzyme (s) is reduced / reduced at least partially as compared to the starting organism: a. R79. A further embodiment of the present invention relates to a method for producing a microorganism of the genus Coryneba cterium with an increased efficiency for metine synthesis wherein • the amount and / or activity of the following enzyme (s) is increased / increased and / or introduce in comparison with the starting organism: 1. R3 in order to produce more GLC-LAC and / or 2. R4 in order to produce more 6-P-Gluconate and / or 3. R5 in order to produce more RIB-5P and / or 4. RIO in order to produce more F6P and GA3P and / or 5. R2 in order to produce more G6P and / or 6. R55 in order to produce more H2S03 and / or 7. R58 in order to produce more H2S and / or 8. R71 in order to produce more M-HPL and / or 9. R72 in order to produce more Methylene-THF and / or . R78 in order to produce Methyl-THF and / or 11. R76 in order to produce more Methylene-THF and / or 12. R70 in order to produce more NADPH and / or 13. R81 in order to produce more NADPH and / or 14. R25 in order to produce more Glu and / or 15. R33 and / or R36 in order to produce more OAA and / or 16. R30 in order to produce more MAL and / or 17. R57 in order to produce more Pyr and / or 18. R80 in order to metabolize sulphate to sulfite and / or 19. R73 in order to metabolize thiosulfate towards sulfur and sulfite and / or 20. R74 in order to metabolize sulfite to sulfide and / or 21. R82 to transport more external H2S20 to the cell and / or 22. R75 in order to produce 10-formyl-THF and / or 23. R76 in order to produce Methylene-THF from 10-formyl-THF and / or 24. R78 in order to produce more Methylene-THF and / or . R77 for the purpose of methyl-sulfhydryl O-Acetyl-homoserine with methanethiol and / or • the amount and / or activity of the following enzyme (s) is reduced / reduced at least partially as compared to the starting organism: 1. Rll in order to produce less F-1,6-BP and / or 2. R19 in order to produce less Pyr and / or 3. R20 in order to produce less Ac-CoA and / or 4. R21 in order to produce less CIT and / or 5. R24 in order to produce less 2-OXO and / or 6. R26 in order to produce less SUCC-CoA and / or 7. R27 in order to produce less SUCC and / or 8. R31 in order to produce less GLYOXY and SUCC and / or 9. R32 in order to produce less BAD and / or 10. R35 in order to produce less PEP and / or 10. R79 in order to produce less THF A further embodiment of the present invention relates to a method for producing a microorganism of the genus Coryneba cterium with an increased efficiency for methionine synthesis, wherein • the amount and / or activity of the following enzyme is increased and / or introduced in comparison with the starting organism: 1. R3 in order to produce more GLC-LAC and / or 2. R4 in order to produce more 6-P-Gluconate and / or 3. R5 in order to produce more RIB-5P and / or 4. RIO in order to produce more F6P and GA3P and / or 5. R2 in order to produce more G6P and 6. R55 in order to produce more H2S03 and / or 7. R58 in order to produce more H2S and 8. R71 in order to produce more M-HPL and / or 9. R72 in order to produce more Methylene-THF and / or . R78 in order to produce Methyl-THF and / or 11. R76 in order to produce more Methylene-THF and / or 12. R70 in order to produce more NADPH and / or 13. R81 in order to produce more NADPH and / or 14. R25 in order to produce more Glu and / or 15. R33 and / or R36 in order to produce more OAA 16. R30 in order to produce more MAL 17. R57 in order to produce more Pyr 18. R80 in order to metabolize sulphate to sulfite and / or 19. R75 in order to produce 10-formyl-THF and / or 20. R76 for the purpose of producing Methylene-THF from 10-formyl-THF and / or 21. R77 for the purpose of methyl-sulfhydrylating O-Acetyl-homoserine with methanethiol and / or the amount and / or activity of the following enzymes are at least partially reduced compared to the starting organism: 1. Rll in order to produce less F-1,6-BP and / or 2. R19 in order to produce less Pyr and / or 3 R20 in order to produce less Ac-CoA and / or 4. R21 in order to produce less CIT and / or 5. R24 in order to produce less 2-OXO and / or 6. R26 in order to produce less SUCC-CoA and / or 7. R27 in order to produce less SUCC and / or 8. R 31 in order to produce less GLYOXY and SUCC and / or 9. R32 in order to produce less MAL and / or 10. R35 in order to produce less PEP and / or 11. R79 in order to produce less THF. Any organism obtained by these methods is also an object of the present invention.

The Coryneba cteri um microorganisms used for these methods can be selected from the group consisting of Coryneba cterium glutami cum ATCC 13032, Corynebacterium acetoglummicum ATCC 15806, Coryneba cteriumum acetoa cidophilum ATCC 13870, Corynebacterium thermoaminogenes FERM BP-1539, and Corynebacteri um melassecola ATCC 17965, Coryneba cteri um gl utami cum KFCC 10065 and Corynebacterium gl utamicum ATCC21608 Corynebacterium gl utamicum DSM 17322 The abbreviations KFCC stand for Korean Federation of Culture Collection, while the abbreviation ATCC stands for the American Type Strain Culture Collection and the abbreviation DSM stands for the Germán Resource Center for Biological Material. Of particular interest are the genetically modified organisms of the genus Corynebacterium, where the metabolic flux through the following pathways is introduced: • glycine cleavage system • transhydrogenase conversion • thiosulfate reductase system • sulfate reductase system • converter system formate • methanethiol converter system.

If a methanethiol converting system is introduced into Corynebacterium, the thiosulfate reductase system and formate converting system may become obsolete. It has been found that these additional path systems mentioned above contribute significantly to the optimal metabolic flux for efficient synthesis of methionine in E. coli (see in the following). According to the theoretical predictions, the inclusion of these metabolic pathways in C. gl utami cum should also increase the efficiency of C. gl utamicum for methionine synthesis. Thus, one aspect of the present invention relates to organisms that have been genetically modified in order to increase metabolic flux through any of the aforementioned pathways. By genetically correcting C. gl utami cum according to the present invention, the metabolic flux in these organisms can be corrected in order to increase the efficiency of methionine synthesis so that these organisms are characterized in that methionine is produced with an efficiency of at least 10%, of at least 20%, of at least 30%, of at least 35%, of at least 40%, of at least 45%, of at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80% or at least 85%.

The present invention is not related, as far as C gl utamicum is concerned, to the strains deactivated in? MabR described in Rey et al. (2003) vide supra.

E. coli An aspect of the present invention relates to a microorganism of the genus Escheri chia () that has been genetically modified in order to increase and / or introduce a metabolic flux through at least one of the following routes compared to the start: phosphotransferase system (PTS) and / or phosphotransferase system (PTS) and / or glycolysis (EMP) and / or tricarboxylic acid cycle (TCA) and / or • glyoxylate (GS) deviation and / or pathway anaplerosis (AP) and / or methionine (MS) synthesis pathway and / or serine / cysteine / glycine system (SCGS) and / or pathway 1 (Pl) and / or • sulfur assimilation (SA) and / or system glycine cleavage (GCS) and / or transhydrogenase conversion (THGC) and / or thiosulfate reductase system (TRS) and / or sulfite reductase system (SRS) and / or • sulfate reductase system (SARS) and / or • formate converter system (FCS) and / or • methanethiol converter system (MCS) and / or • serine / cysteine / glycine synthesis ( SCGS). These microorganisms with increased methionine synthesis efficiency are optionally also characterized by at least a decreased metabolic flux through at least one of the following pathways compared to the start that can also be achieved by genetic modification: • pentose pathway phosphate (PPP) • synthesis of methionine (MS) and / or • via 3 (P3) and / or • via 4 (P4) and / or • via 7 (P7) and / or • via 8 (P8). In some modalities, the metabolic flow through PPP may not decrease but may increase. In addition to the microorganisms of the genus Escherichia mentioned in the above, the present invention also relates to a method for producing microorganisms of the genus Escherichia with increased production efficiency of methionine comprising the following steps: • increase and / or introduce the metabolic flux to through at least one of the following pathways compared to the start by genetic modification of the organism: phosphotransferase system (PTS) and / or glycolysis (EMP) and / or tricarboxylic acid cycle (TCA) and / or deviation of glyoxylate (GS) and / or anaplerosis pathway (AP) and / or methionine synthesis pathway (MS) and / or serine / cysteine / glycine system (SCGS) and / or • via 1 (Pl) and / or assimilation of sulfur (SA) and / or glycine cleavage system (GCS) and / or transhydrogenase conversion (THGC) and / or thiosulfate reductase system (TRS) and / or • sulfite reductase system (SRS) and / or system of sulfate reductase (SARS) and / or formate converting system (FCS) and / or methanethiol converting system (MCS) and / or serine / cysteine / glycine synthesis (SCGS) and / or • at least partially decrease the metabolic flux through less one of the following ways compared to the start by the genetic modification of the organism: • via pentose phosphate (PPP) • path 3 (P3) and / or • via 4 (P4) and / or • via 7 (P7) ) and / or • R19 and / or • R35 and / or • R79. Additional aspects of the present invention are the methods for producing a microorganism of the genus Escheri chia with an increased efficiency for methionine synthesis wherein with respect to PTS, the amount and / or activity of the following enzyme is increased in comparison with the organism of start: a. Rl in order to produce more G6P; and / or with respect to EMO, the amount and / or activity of the following enzyme (s) is increased / increased and / or introduced compared to the starting organism: a. R2 in order to produce more F6P and / or b. Rll in order to produce more F-1,6-BP and / or c. R13 in order to produce more DHAP and GA3P and / or d. R14 in order to produce more GA3P and / or e. R15 in order to produce more 1,3-PG and / or f. R16 in order to produce more 3-PG and / or g. R17 in order to produce more 2-PG and / or h. R18 in order to produce more PEP and / or i. R19 in order to produce more Pyr; and / or with respect to TCA, the amount and / or activity of the following enzyme (s) is increased / increased and / or introduced compared to the starting organism: a. R20 in order to produce more Ac-CoA and / or b. R21 in order to produce more CIT and / or c. R22 in order to produce more Cis-ACO and / or d. R23 in order to produce more ICI and / or e. R24 in order to produce more 2-0X0 and / or f. R26 in order to produce more SUCC-CoA and / or g. R27 in order to produce more SUCC and / or h. R28 in order to produce more FUM and / or i. R29 in order to produce more MAL and / or j. R30 in order to produce more OAA; and / or with respect to AP, the amount and / or activity of the following enzyme is increased and / or introduced compared to the starting organism: a. R33 in order to produce more OAA and / or with respect to MS, the amount and / or activity of the following enzyme is increased and / or introduced compared to the starting organism: a. R37 in order to produce more Asp and / or b. R39 in order to produce more HOM and / or c. R40 in order to produce more 0-AC-HOM and / or d. R46 in order to produce more CYSTA and / or e. R47 in order to produce more ASP-P and / or f. R48 in order to produce more ASP-SA and / or g. R49 in order to produce more HOMOCYS and / or h. R52 in order to produce more MET and / or i. R53 in order to produce more METex and / or j. R54 in order to produce more HOMOCYS and / or with respect to SCGS, the amount and / or activity of the following enzyme is increased and / or introduced compared to the starting organism: a. R38 in order to produce more M-THF and * Glycine ex and / or b. R44 in order to produce more O-AC-SER and / or c. R45 in order to produce more CYS and / or with respect to GS, the amount and / or activity of the following enzyme (s) is increased / increased and / or introduced compared to the starting organism: a. R21 in order to produce more CIT and / or b. R22 in order to produce more Cis-ACO and / or c. R23 in order to produce more ICI and / or d. R31 in order to produce more GLYOXY and SUCC and / or e. R32 in order to produce more MAL and / or f. R28 in order to produce more FUM and / or g. R29 in order to produce more MAL and / or h. R30 in order to produce more OAA; and / or with respect to Pl, the amount and / or activity of the following enzymes is increased / increased and / or introduced compared to the starting organism: R25 in order to produce more Glu; and / or with respect to SA, the amount and / or activity of the following enzymes is increased / increased and / or introduced compared to the starting organism: a. R55 in order to produce more H2S03 and / or b. R58 in order to produce more H2S; and / or with respect to GCS, the amount and / or activity of the following enzymes is increased / increased and / or introduced compared to the starting organism: a. R71 in order to produce more M-HPL and / or b. R72 in order to produce more methylene-THF and / or c. R78 in order to produce more Methyl-THF and / or with respect to claiming THGC, the amount and / or activity of the following enzyme is increased and / or introduced compared to the starting organism: a. R70 in order to produce more NADPH from NADH and / or b. R81 in order to produce more NADPH from NADH and / or with respect to TRS, the amount and / or activity of the following enzymes is increased / increased and / or introduced compared to the starting organism: a. R73 in order to metabolize thiosulfate to sulfur and sulphite and / or b. R82 to transport more external H2S203 towards the cell and / or with respect to SRS, the amount and / or activity of the following enzyme is increased and / or introduced compared to the starting organism: a. R74 in order to metabolize sulfite to sulfide and / or with respect to SARS, the amount and / or activity of the following enzyme is increased and / or introduced compared to the starting organism: a. R80 in order to metabolize sulfate to sulfite and / or with respect to FCS, the amount and / or activity of the following enzymes is increased / increased and / or introduced compared to the starting organism: a. R75 in order to produce 10-formyl-THF and / or b. R76 in order to produce Methylene-THF from 10-formyl-THF and / or c. R78 in order to produce more Methyl-THF with respect to MCS, the amount and / or activity of the following enzyme is increased and / or introduced compared to the starting organism: a. R77 for the purpose of methyl sulfhydryl O-Acetyl-homoserine with methanethiol and / or with respect to SCGS, the amount and / or activity of the following enzyme is increased in comparison with the starting organism: a. R44 in order to produce more O-Ac-SER and / or b. R45 in order to produce more CYS; and / or with respect to PPP, the amount and / or activity of the following enzyme (s) is increased / increased as compared to the starting organism: a. R3 in order to produce more GLC-LAC and / or b. R4 in order to produce more 6-P-Gluconate and / or c. R5 in order to produce more RIB-5P and / or d. R6 in order to produce more XYL-5P and / or e. R7 in order to produce more RIBO-5P and / or f. R8 in order to produce more S7P and GA3P and / or g. R9 in order to produce more E-4p and F6P and / or h. RIO in order to produce less F6P and GA3P and / or i. R2 in order to produce 1 more G6P; and / or with respect to PPP, in some embodiments the amount and / or activity of the following enzyme (s) may also be at least partially reduced compared to the starting organism: j. R3 in order to produce less GLC-LAC and / or k. R4 in order to produce less 6-P-Gluconate and / or 1. R5 in order to produce less RIB-5P and / or m. R6 in order to produce less XYL-5P and / or n R7 in order to produce less RIB0-5P and / or R8 in order to produce less S7P and GA3P and / or p. R9 in order to produce less E-4p and F6P and / or q. RIO in order to produce less F6P and GA3P and / or r. R2 in order to produce less G6P; and / or with respect to P3, the amount and / or activity of the following enzyme (s) is at least reduced / reduced compared to the starting organism: a. R56; and / or with respect to P4, the amount and / or activity of the following enzyme (s) is at least reduced / reduced compared to the starting organism: a. R62; and / or with respect to P7, the amount and / or activity of the following enzyme (s) is at least reduced / reduced compared to the starting organism: a. R61. the amount and / or activity of the following enzyme (s) is at least reduced / reduced compared to the starting organism: a. R19 in order to produce less pyruvate; and / or b. R35 in order to produce less PEP; and / or c. R79 in order to produce less tetrahydrofolate. A further embodiment of the invention with respect to the genus Escheri chia relates to a method for producing Escherichia microorganisms with increased efficiency of methionine synthesis, wherein the amount and / or activity of the following enzymes is increased and / or introduced in comparison with the starting organism: 1. Rl in order to produce more G6P and / or 2. R2 in order to produce more F6P and / or 3. Rll in order to produce more F-1,6-BP and / or 4. R19 in order to produce more Pyr and / or 5. R20 in order to produce more Ac-CoA and / or 6. R21 in order to produce more CIT and / or 7. R24 in order to produce plus 2-OXO and / or 8. R26 in order to produce more SUCC-CoA and / or 9. R31 in order to produce more GLYOXY and SUCC and / or 10. R32 in order to produce more MAL and / or 11. R25 in order to produce more Glu and / or 12. R55 in order to produce more H2S03 and / or 13. R58 in order to produce more H2S and / or 14. R71 in order to produce more M- HPL and / or 15. R72 with in order to produce more M-THF and / or 16. R78 in order to produce more Methyl-THF and / or 17. R76 in order to produce more Methylene-THF and / or 18. R70 in order to produce more NADPH and / or 19. R81 in order to produce more NADPH and / or 20. R80 in order to metabolize sulfate to sulfite and / or 21. R73 in order to metabolize thiosulfate to sulfide and sulphite and / or 22. R82 to transport more external H2S03 to the cell and / or 23. R74 in order to metabolize sulfite to sulfide and / or 24. R75 in order to produce 10-formyl-THF and / or 25. R76 for the purpose of producing Methylene-THF from 10-formyl-THF and / or 26. R77 for the purpose of methyl-sulfhydrylating O-Acetyl-homoserine with methanethiol and / or 27. R44 in order to produce more O -Ac-SER and / or 28. R45 in order to produce more CYS; • the amount and / or activity of the following enzyme (s) is reduced / reduced at least partially compared to the starting organism: 1. R19 in order to produce less pyruvate; and / or 2. R35 in order to produce less PEP; and / or 3. R79 in order to produce less tetrahydrofolate. The microorganism of the genus Escheri chia that can be obtained by any of the methods mentioned above is selected from the group comprising for example Escherichia coli.

In some modalities that relate to organisms such as E. coli and C. gl utami cum, the metabolic flux is generated by overexpression of the following enzymatic activities: R37, R38, R39, R40, R44, R45, R46, R47, R48, R49, R52, R53, R54 and / or R58. The organisms E. coli and C. gl u tamicum in which any combination of the R-numbers mentioned above or any of the genes that are part of these catalytic activities are overexpressed, also form an object of the invention. Organizations such as E. coli and C. gl utami cum in which any combination of R70, R81, R71 / R72, R73, R82, R74, R75, R76, R77, R78, and / or R80 together with R37, R38, R39, R40, R44 , R45, R46, R47, R48, R49, R52, R53, R54 and / or R58 or the genes that are part of these catalytic activities are overexpressed, they also form an object of the invention. Organizations such as E. coll and C. gl utamicum in which an enzymatic activity of the group consisting of R70, R81, R71 / R72, R73, R82, R74, R75, R76, R77, R78, and R80 together with either R37, R38, R39 , R40, R44, R45, R46, R47, R48, R49, R52, R53, R54 and / or R58 or the genes that are part of these catalytic activities are overexpressed, they also form an object of the invention. Organizations such as E. col i and C. gl utamicum in which two enzymatic activities of the group consisting of R70, R81, R71 / R72, R73, R82, R74, R75, R76, R77, R78, and R80 together with either of R37, R38, R39 , R40, R44, R45, R46, R47, R48, R49, R52, R53, R54 and / or R58 or the genes that are part of these catalytic activities are overexpressed, they also form an object of the invention. Organisms such as E. coli and C. gl utamicum in which three enzymatic activities of the group consisting of R70, R81, R71 / R72, R73, R82, R74, R75, R76, R77, R78, and R80 together with any of R37, R38, R39, R40, R44, R45, R46, R47, R48, R49, R52, R53, R54 and / or R58 or the genes that are part of these catalytic activities are overexpressed, they also form an object of the invention. Organizations such as E. coli and C. gl utami cum in which four enzymatic activities of the group consisting of R70, R81, R71 / R72, R73, R82, R74, R75, R76, R77, R78, and R80 together with either of R37, R38, R39, R40, R44, R45, R46, R47, R48, R49, R52, R53, R54 and / or R58 or the genes that are part of these catalytic activities are overexpressed, they also form an object of the invention. Organizations such as E. coli and C. glutamicum in which five enzymatic activities of the group consisting of R70, R81, R71 / R72, R73, R82, R74, R75, R76, R77, R78, and R80 together with either R37, R38, R39, R40, R44, R45, R46, R47, R48, R49, R52, R53, R54 and / or R58 or the genes that are part of these catalytic activities are overexpressed, they also form an object of the invention.

Organisms such as E. coli and C. gl utamicum in which six enzymatic activities of the group consisting of R70, R81, R71 / R72, R73, R82, R74, R75, R76, R77, R78, and R80 together with any of R37, R38, R39, R40, R44, R45, R46, R47, R48, R49, R52, R53 , R54 and / or R58 or the genes that are part of these catalytic activities are overexpressed, they also form an object of the invention. Organizations such as E. coli and C. gl utamicum in which seven enzymatic activities of the group consisting of R70, R81, R71 / R72, R73, R82, R74, R75, R76, R77, R78, and R80 together with either of R37, R38, R39 , R40, R44, R45, R46, R47, R48, R49, R52, R53, R54 and / or R58 or the genes that are part of these catalytic activities are overexpressed, they also form an object of the invention. Organizations such as E. coli and C. gl utamicum in which eight enzymatic activities of the group consisting of R70, R81, R71 / R72, R73, R82, R74, R75, R76, R77, R78, and R80 together with either of R37, R38, R39 , R40, R44, R45, R46, R47, R48, R49, R52, R53, R54 and / or R58 or the genes that are part of these catalytic activities are overexpressed, they also form an object of the invention. Organizations such as E. coli and C. gl utamicum in which nine enzymatic activities of the group consisting of R70, R81, R71 / R72, R73, R82, R74, R75, R76, R77, R78, and R80 together with either of R37, R38, R39 , R40, R44, R45, R46, R47, R48, R49, R52, R53, R54 and / or R58 or the genes that are part of these catalytic activities are overexpressed, they also form an object of the invention. Organizations such as E. coli and C. gl utamicum in which at least one enzymatic activity of the group consisting of R70, R81, R71 / R72, R73, R82, R74, R75, R76, R77, R78 and R80 together with any of R37, R38 , R39, R40, R44, R45, R46, R47, R48, R49, R52, R53, R54 and / or R58 or the genes that are part of these catalytic activities are overexpressed and also at least one enzyme activity of the group consisting of of R19, R35 and R79 decreases, they also form an object of the invention. Organizations such as E. coli and C. gl utami cum in which at least two enzymatic activities of the group consisting of R70, R81, R71 / R72, R73, R82, R74, R75, R76, R77, R78 and R80 together with either of R37, R38, R39, R40, R44, R45, R46, R47, R48, R49, R52, R53, R54 and / or R58 or the genes that are part of these catalytic activities are overexpressed and also at least one enzyme activity of the group that consists of R19, R35 and R79 decreases, they also form an object of the invention. Organizations such as E. coli and C. gl utamicum in which at least three enzymatic activities of the group consisting of R70, R81, R71 / R72, R73, R82, R74, R75, R76, R77, R78 and R80 together with either of R37, R38 , R39, R40, R44, R45, R46, R47, R48, R49, R52, R53, R54 and / or R58 or the genes that are part of these catalytic activities are overexpressed and also at least one enzyme activity of the group consisting of of R19, R35 and R79 decreases, they also form an object of the invention. Organizations such as E. coli and C. gl utamicum in which at least four enzymatic activities of the group consisting of R70, R81, R71 / R72, R73, R82, R74, R75, R76, R77, R78 and R80 together with either of R37, R38 , R39, R40, R44, R45, R46, R47, R48, R49, R52, R53, R54 and / or R58 or the genes that are part of these catalytic activities are overexpressed and also at least one enzyme activity of the group consisting of of R19, R35 and R79 decreases, they also form an object of the invention. Organizations such as E. coli and C. gl utamicum in which at least five enzymatic activities of the group consisting of R70, R81, R71 / R72, R73, R82, R74, R75, R76, R77, R78 and R80 together with either of R37, R38 , R39, R40, R44, R45, R46, R47, R48, R49, R52, R53, R54 and / or R58 or the genes that are part of these catalytic activities are overexpressed and also at least one enzyme activity of the group consisting of of R19, R35 and R79 decreases, they also form an object of the invention. Organizations such as E. coli and C. gl utamicum in which at least six enzymatic activities of the group consisting of R70, R81, R71 / R72, R73, R82, R74, R75, R76, R77, R78 and R80 together with any of R37, R38, R39, R40, R44, R45, R46, R47, R48, R49, R52, R53, R54 and / or R58 or the genes that are part of these catalytic activities are overexpressed and also at least one enzymatic activity of the group consisting of R19, R35 and R79 decreases, they also form an object of the invention. Organizations such as E. coli and C. gl utami cum in which at least seven enzymatic activities of the group consisting of R70, R81, R71 / R72, R73, R82, R74, R75, R76, R77, R78 and R80 together with either of R37, R38, R39, R40, R44, R45, R46, R47, R48, R49, R52, R53, R54 and / or R58 or the genes that are part of these catalytic activities are overexpressed and also at least one enzyme activity of the group that consists of R19, R35 and R79 decreases, they also form an object of the invention. Organizations such as E. coli and C. gl utamicum in which at least eight enzymatic activities of the group consisting of R70, R81, R71 / R72, R73, R82, R74, R75, R76, R77, R78 and R80 together with either of R37, R38 , R39, R40, R44, R45, R46, R47, R48, R49, R52, R53, R54 and / or R58 or the genes that are part of these catalytic activities are overexpressed and also at least one enzyme activity of the group consisting of of R19, R35 and R79 decreases, they also form an object of the invention.

Organisms such as E. coli and C. gl utami cum in which at least nine enzymatic activities of the group consisting of R70, R81, R71 / R72, R73, R82, R74, R75, R76, R77, R78 and R80 together with any of R37, R38, R39, R40, R44, R45, R46, R47, R48, R49, R52, R53, R54 and / or R58 or the genes that are part of these catalytic activities are overexpressed and also at least one Enzymatic activity of the group consisting of R19, R35 and R79 decreases, they also form an object of the invention. Organizations such as E. coli and C. gl utamicum in which at least one enzymatic activity of the group consisting of R70, R81, R71 / R72, R73, R82, R74, R75, R76, R77, R78 and R80 together with either of R37, R38 , R39, R40, R44, R45, R46, R47, R48, R49, R52, R53, R54 and / or R58 or the genes that are part of these catalytic activities are overexpressed and also at least two enzymatic activities of the group consisting of of R19, R35 and R79 decrease, they also form an object of the invention. Organizations such as E. coli and C. gl utamicum in which at least one enzymatic activity of the group consisting of R70, R81, R71 / R72, R73, R82, R74, R75, R76, R77, R78 and R80 together with either of R37, R38 , R39, R40, R44, R45, R46, R47, R48, R49, R52, R53, R54 and / or R58 or the genes that are part of these catalytic activities are overexpressed and also at least three enzymatic activities of the group consisting of of R19, R35 and R79 decrease, they also form an object of the invention. In a preferred embodiment, the invention relates to a C. gl utamicum organism in which the metabolic flux through one of the following routes is introduced and / or increased for example by genetic modification as described above: FCS or GCS or MCS or TRS or THGC. In another preferred embodiment of the invention, these organisms are further grown using Sulfur or Thiosulfate as external sources of sulfur. In another preferred embodiment, the invention relates to a C. glutami cum organism in which the metabolic flux through the following pathways is introduced and / or increased: FCS and GCS, FCS and MCS, FCS and TRS, or FCS and THGC. In another preferred embodiment of the invention, these organisms are further grown using Sulfur or Thiosulfate as external sources of sulfur. In another preferred embodiment, the invention relates to a C. gl utamicum organism in which the metabolic flux through the following pathways is introduced and / or increased: FCS and GCS and TRS, FCS and GCS and TRS, or FCS and GCS and THGC. In another preferred embodiment of the invention, these organisms are further grown using Sulfur or Thiosulfate as external sources of sulfur. In another preferred embodiment, the invention relates to a C. glutamium cum organism in which the metabolic flux through the following pathways is introduced and / or increased: FCS and GCS and MCS and TRS, or FCS and GCS and MCS and THGC. In another preferred embodiment of the invention, these organisms are further grown using Sulfur or Thiosulfate as external sources of sulfur. In another preferred embodiment, the invention relates to a C. gl utamicum organism in which the metabolic flux through the following pathways is introduced and / or increased: FCS and GCS and MCS and TRS and THGC. In another preferred embodiment of the invention, these organisms are further grown using Sulfur or Trosulfate as external sources of sulfur. In another preferred embodiment, the invention relates to a C. gl utamicum organism in which the metabolic flux through the following pathways is introduced and / or increased: FCS and MCS and TRS, or FCS and MCS and THGC. In another preferred embodiment of the invention, these organisms are further grown using Sulfur or Thiosulfate as external sources of sulfur. In another preferred embodiment, the invention relates to a C. gl utami cum organism in which the metabolic flux through the following pathways is introduced and / or increased: FCS and MCS and TRS and THGC. In another preferred embodiment of the invention, these organisms are further grown using Sulfur or Thiosulfate as external sources of sulfur. In another preferred embodiment, the invention relates to a C. gl utami cum organism in which the metabolic flux through the following pathways is introduced and / or increased: FCS and TRS and THGC. In another preferred embodiment of the invention, these organisms are further grown using Sulfur or Thiosulfate as external sources of sulfur. In another preferred embodiment, the invention relates to a C. gl utamicum organism in which the metabolic flux through the following pathways is introduced and / or increased: FCS and MCS and TRS, or FCS and MCS and THGC. In another preferred embodiment of the invention, these organisms are further grown using Sulfur or Thiosulfate as external sources of sulfur. In another preferred embodiment, the invention relates to an organism C glutamine cum in which the metabolic flux through the following pathways is introduced and / or increased: FCS and MCS and TRS and THGC. In another preferred embodiment of the invention, these organisms are further grown using Sulfur or Thiosulfate as external sources of sulfur. In another preferred embodiment, the invention relates to a C. gl utami cum organism in which the metabolic flux through the following pathways is introduced and / or increased: MCS and TRS, or MCS and THGC. In another preferred embodiment of the invention, these organisms are further grown using Sulfur or Thiosulfate as external sources of sulfur. In another preferred embodiment, the invention relates to a C. gl utami cum organism in which the metabolic flux through the following pathways is introduced and / or increased: MCS and TRS and THGC. In another preferred embodiment of the invention, these organisms are further grown using Sulfur or Thiosulfate as external sources of sulfur. In another preferred embodiment, the invention relates to a C. gl utamicum organism in which the metabolic flux through the following pathways is introduced and / or increased: TRS and THGC. In another preferred embodiment of the invention, these organisms are further grown using Sulfur or Thiosulfate as external sources of sulfur. For genetic manipulation in the case of GCS, the expression of R71 and / or R72 may be increased. In the case of THGS, the expression of R70 and / or R81 may be increased. In the case of TRS, the expression of R73, R45a, R49 and / or R82 may be increased. For MCS, the expression of R77 may be increased. In the case of FCS, the expression of R75, R76 and / or R78 may be increased. A preferred embodiment of the invention is represented in Figure 10. In this embodiment, an organism is represented in which the metabolic flux through the following routes is increased and / or introduced compared to the starting organism, for example by means of of the aforementioned genetic manipulations: FCS and GCS and MCS and TRS and THGC. Concomitantly, the use of Sulfur and Thiosulfate as sources of Sulfur is considered. The organisms of the present invention may preferably comprise a microorganism of the genus Corynebacterium, particularly Corynebacterium acetoacidophilum, C. acetoglutamicum, C. efficiens, C. jejeki, C. acetophilum, C. ammoniagenes, C. glutamicum, C. lilium, C. nitrilophilus. or C. spec. Organisms according to the present invention also comprise members of the genus Brevibacterium, such as Brevibacterium harmoniagenes, Brevibacterium botanicum, B. divaraticum, B. flavam, B. healil, B. ketoglutamicum, B. ketosoreductum, B. lactofermentum, B. linens , B. paraphinolyticum and B. spec. As for the genus Escherichia, the present invention concerns eg E. col. As stipulated in the above, the metabolic flux through a specific reaction or specific metabolic pathway can be modified by increasing or decreasing the amount and / or activity of the enzymes that catalyze the respective reactions. With respect to increasing the amount and / or activity of an enzyme, all methods known in the art for increasing the amount and / or activity of a protein in a host such as the organisms mentioned in the above can be used.

Increment or enter the amount and / or activity With respect to increasing the quantity, two basic scenarios can be differentiated. In the first scenario, the amount of the enzyme is increased by the expression of an exogenous version of the respective pin. In the other scenario, the expression of the endogenous pin is increased by influencing the activity of the promoter and / or enhancer element and / or other regulatory activities such as phosphorylation, sumolization, ubiquitination etc. that regulate the activities of the respective pins at a transcriptional, translational or post-translational level. In addition to simply increasing the amount, for example, of the enzymes in Table 1, the activity of the pins can be increased by using enzymes that can carry specific mutations that allow for an increased activity of the enzyme. Such mutations can, for example, inactivate regions of an enzyme that are responsible for feedback inhibition. By mutating these, for example by introducing non-conservative mutations, the enzyme can no longer allow the regulation of feedback and thus the activity of the enzyme can not be down-regulated if more product was produced. Mutations can be introduced into the endogenous copy of the enzyme, or they can be provided by over-expressing a corresponding mutant form of the exogenous enzyme. Such mutations may comprise point mutations, deletions or insertions. Point mutations can be conservative or non-conservative. Additionally, deletions may comprise only two or three amino acids up to complete domains of the respective pin. In this way, the increase of the activity and the amount of a pin can be achieved by different routes, for example by disconnecting inhibitory regulatory mechanisms at the level of transcription, translation, and pin or by increasing the gene expression of a nucleic acid. encoding these pins compared to the start, for example by inducing the endogenous R3 gene or by introducing nucleic acids encoding R3. In one embodiment, the increase in enzyme activity and amount, respectively, compared to the start is achieved by an increase in gene expression of a nucleic acid encoding such enzymes. The sequences can be obtained from the respective database, for example in NCBI (http://www.ncbi.nlm.nih.gov/), EMBL (http://www.embl.org), or Expasy (http: //www.expasy.org/). Examples are given in Table 1. In a further embodiment, the increase in the amount and / or activity of the enzymes of Table 1 is achieved by introducing nucleic acids encoding the enzymes of Table 1 into the organism, preferably C. gl utami cum E. coli. In principle, any pin from different organisms with an enzymatic activity of the pins listed in Table 1, can be used. With the genomic sequences of the nucleic acids of such enzymes from eukaryotic sources containing introns, nucleic acid sequences already processed as the corresponding cDNAs should be used in case the host organism is unable or unable to become capable of splicing the corresponding mRNAs. All nucleic acids mentioned in the description can be, for example, an RNA, DNA or cDNA sequence. In a method according to the present invention, to produce organisms with increased efficiency of methionine synthesis, a nucleic acid sequence encoding one of the enzymes, regulated by feedback or independent of feedback, functional or non-functional defined in the foregoing, it is transferred to a microorganism such as C. gl utamicum or E. coli. , respectively. This transfer leads to an increase in the expression of the enzyme, respectively, and corresponding to more metabolic flux through the desired reaction pathway. In accordance with the present invention, increasing or introducing the amount and / or activity of a protein typically comprises the following steps: a) production of a vector comprising the following nucleic acid sequences, preferably DNA sequences, in 5'-3 'orientation: a sequence functional promoter in the organisms of the invention operably linked thereto, a DNA sequence encoding a protein of Table 1 or functional equivalent parts thereof - a functional terminator sequence in the organisms of the invention b) transfer of the vector of step a) to the organisms of the invention such as C. gl utamicum or E. coli and, optionally, integration into the respective genomes. When functionally equivalent parts of the enzymes are mentioned within the scope of the present invention, fragments of nucleic acid sequences encoding enzymes of Table 1 are implicated, whose expression still leads to proteins having the respective protein length enzymatic activity. total. According to the present invention, the non-functional enzymes have the same nucleic acid sequences and amino acid sequences, respectively, as functional enzymes and functionally equivalent parts thereof, respectively, but have, in some positions, point mutations, insertions or deletions of nucleotides or amino acids, which have the effect that non-functional enzymes are not capable, or only to a very limited extent, of catalyzing the respective reaction. These non-functional enzymes can not be intermixed with enzymes that are still capable of catalyzing the respective reaction, but which are no longer regulated by feedback. The non-functional enzymes also comprise such enzymes of Table 1 which carry point mutations, insertions, or deletions at the level of nucleic acid sequences or amino acid sequence level and are not capable, or however, of interacting with physiological associates of union of enzymes. Such physiological binding partners comprise, for example, the respective substrates. What the non-functional mutants are unable to do is to catalyze a reaction that the wild-type enzyme can, from which the mutant is derived. According to the present invention, the term "non-functional enzyme" does not comprise such proteins that do not have essential sequence homology with the respective functional enzymes at the amino acid and nucleic acid levels, respectively. Proteins incapable of catalyzing the respective reactions and having no essential homology of sequences with the respective enzyme are therefore, by definition, not implied by the term "non-functional enzyme" of the present invention. Non-functional enzymes, within the scope of the present invention, are also referred to as inactivated or inactive enzymes. Therefore, the non-functional enzymes of Table 1 according to the present invention, carrying the point mutations, insertions, and / or eliminations mentioned in the foregoing, are characterized by an essential homology of sequences with the wild-type enzymes. of Table 1 according to the present invention or functionally equivalent parts thereof. In accordance with the present invention, it is generally understood that substantial sequence homology indicates that the nucleic acid sequence or amino acid sequence, respectively, of a DNA molecule or a protein, respectively, is at least 25%, by at least 30%, at least 40%, preferably at least 50%, additionally at least 60%, preferably also at least 70%, also preferably at least 80%, particularly preferably at least 90% %, in particular at least 95% and more preferably at least 98% identical to nucleic acid sequences or amino acid sequences is preferred., respectively, of the proteins of Table 1 or functionally equivalent parts thereof. The identity of two proteins is understood to be the identity of the amino acids over the respective full length of the protein, in particular the identity calculated by comparison with the aid of the Lasergene software by DNA Star, Inc., Madison, Wisconsin (USA). ) applying the CLUSTAL method (Higgins et al., (1989), Comput. Appl. Biosci., 5 (2), 151). The homologies can also be calculated with the help of the Lasergene software by DNA Star, Inc., Madison, Wisconsin (USA) applying the CLUSTAL method (Higgins et al., (1989), Comput. Appl. Biosci., 5 (2), 151). The identity of DNA sequences must be understood accordingly. The method mentioned in the above can be used to increase the expression of DNA sequences encoding enzymes regulated by feedback or independent of feedback, functional or non-functional of Table 1 or functionally equivalent parts thereof. The use of such vectors comprising regulatory sequences, such as promoter and terminator sequences, is known to the person skilled in the art. Additionally, the person skilled in the art knows how a vector of step a) can be transferred to organisms such as C. glutami cum or E. coli and what properties a vector must have to be able to integrate into its genomes. If the enzyme content in an organism such as C. gl utamicum is increased by transferring a nucleic acid encoding an enzyme from another organism, such as E. coli, it is advisable to transfer the amino acid sequence encoded by the nucleic acid sequence for example from E. coli by reverse translation of the polypeptide sequence according to the genetic code towards a nucleic acid sequence comprising mainly those codons, which are used more often due to the use of specific codons of the organism. The use of codons can be determined by means of computer evaluations of other known genes of the relevant organisms. According to the present invention, an increase in gene expression and activity, respectively, of a nucleic acid encoding an enzyme of Table 1 is also understood as the manipulation of the expression of the respective endogenous enzymes endogenous to an organism , in particular of C. gl utamicum or E. coli This can be achieved, for example, by altering the promoter DNA sequence for the genes encoding these enzymes. Such alteration, which causes an altered, preferably increased expression rate of these enzymes, can be achieved by the elimination or insertion of DNA sequences. An alteration of the promoter sequence of endogenous genes usually causes an alteration of the expressed amount of the gene and therefore also an alteration of the detectable activity in the cell or in the organism. Additionally, an altered and increased expression, respectively, of an endogenous gene can be achieved by a regulatory protein, which does not occur in the transformed organism, and which interacts with the promoter of these genes. Such a regulator can be a chimeric protein consisting of a DNA binding domain and a transcription activating domain, for example as described in WO 96/06166. An additional possibility to increase the activity and content of endogenous genes is to over-regulate transcription factors involved in the transcription of the endogenous genes, for example by means of overexpression. The measures for overexpression of transcription factors are known to the person skilled in the art and are also described for the enzymes of Table 1 within the scope of the present invention. Additionally, an alteration of endogenous gene activity can be achieved by site-directed mutagenesis of the copies of the endogenous gene. An alteration of the endogenous genes encoding the enzymes in Table 1 can also be achieved by influencing the post-translational modifications of the enzymes. This can happen for example by regulating the activity of enzymes such as kinases or phosphatases involved in the post-translational modification of the enzymes by means of corresponding measures such as overexpression or gene silencing. In another embodiment, an enzyme can be improved in efficiency, or its allosteric control region destroyed so as to prevent feedback inhibition of compound production. Similarly, a degradative enzyme can be removed or modified by substitution, elimination, or addition so that its degradative activity is reduced to the desired enzyme of Table 1 without impairing the viability of the cell. In each case, the yield or overall production rate of one of these desired fine chemicals can be increased. It is also possible that such alterations in the protein and nucleotide molecules of Table 1 may improve the production of other fine chemicals such as other sulfur containing compounds such as cysteine or glutathione, other amino acids, vitamins, cofactors, nutritional drugs, nucleotides, nucleosides , and trehalose. The metabolism of any compound is necessarily intertwined with other biosynthetic and degradative pathways within the cell, and the necessary cofactors, intermediates, or substrates in one pathway are likely to be supplied or bound in another way. Thus, by modulating the activity of one or more of the proteins in Table 1, the production or efficiency of the activity of another biosynthetic or degradative pathway of fine chemicals in addition to those that lead to methionine, can be impacted. The expression and function of the enzymes can also be regulated based on the cellular levels of a compound of a different metabolic process, and the cellular levels of molecules necessary for basic growth, such as amino acids and nucleotides, can critically affect the viability of the microorganism in large-scale cultivation. In this way, the modulation of the biosynthesis enzymes of an amino acid from Table 1 so that they can no longer respond to inhibition by feedback or so that they are improved in efficiency or renewal should result in a superior metabolic flux through the methionine production pathways. The theoretical method of the invention will help to incorporate the effects of these nutrients, metabolites, etc. in the model organisms and in this way will provide a valuable guide to the metabolic pathways that must be genetically modified to increase the efficiency of methionine synthesis. These strategies mentioned above to increase or introduce the amount and / or activity of the enzymes in Table 1 does not mean that they should be limiting; Variations in these strategies will be readily apparent to someone with ordinary skill in the art.

Reduce the amount and / or activity of enzymes To reduce the amount and / or activity of any of the enzymes in Table 1, various strategies are also available. The expression of the endogenous enzymes of Table 1 for example can be regulated by the expression of aptamers that specifically bind to the promoter sequences of the genes. Depending on the binding of the aptamers to stimulate or repress promoter regions, the amount, and thus in this case, the activity of the enzymes of Table 1, is increased or reduced. The aptamers can also be designed to bind specifically to the enzymes themselves and to reduce the activity of the enzymes for example by binding to the catalytic center of the respective enzymes. The expression of aptamers is usually achieved by vector-based overexpression (see above) and, as well as the design and selection of aptamers, is well known to the person skilled in the art (Famulok et al., (1999) Curr. Top Microbiol Immunol, 243, 123-36). Additionally, a decrease in the amount and activity of the endogenous enzymes of Table 1 can be achieved by means of various experimental measures, which are well known to the person skilled in the art. These measures are usually summarized under the term "gene silencing". For example, the expression of an endogenous gene can be silenced by transferring a vector mentioned in the above, having a DNA sequence encoding the enzyme or parts thereof in antisense order, to organisms such as C. gl utamicum and E. . coli This is based on the fact that the transcription of such a vector in the cell leads to an RNA, which can hybridize with the mRNA transcribed by the endogenous gene and therefore prevents its translation. Regulatory sequences operably linked to a nucleic acid cloned in the antisense orientation which direct the continuous expression of the antisense RNA molecule in a variety of cell types, for example viral promoters and / or enhancers, can be chosen, or regulatory sequences can be chosen which direct the specific expression of the tissue or specify the cellular, constitutive type of antisense RNA. The antisense expression vector can be in the form of a recombinant plasmid, phagemid or attenuated virus in which the antisense nucleic acids are produced under the control of a high efficiency regulatory region, whose activity can be determined by the cell type in which the Vector is introduced. For a discussion of the regulation of gene expression using antisense genes see Weintraub, H. et al., Antisense RNA as a molecular tool for genetic analysis, Rewiews-Tren s in Genetics, Vol. 1 (1) 1986. In principle, the antisense strategy can be coupled with a ribozyme method. Ribozymes are catalytically active RNA sequences, which, if coupled to the antisense sequences, catalytically cleave the target sequences (Tanner et al., (1999) FEMS Microbiol Rev. 23 (3), 257-75). This can intensify the efficiency of an antisense strategy. In plants, gene silencing can be achieved by RNA interference or a process known as co-suppression. Additional methods are the introduction of nonsense mutations in the endogenous gene by introducing RNA / DNA oligonucleotides into the organism (Zhu et al., (2000) Na t Biotechnol.18 (5), 555-558) or generating mutants deactivated with the help of homologous recombination (Hohn et al., (1999) Proc. Na ti. Acad. Sci. USA 96, 8321-8323.). To create a homologous recombinant microorganism, a vector is prepared that contains at least a portion of the gene encoding an enzyme of Table 1 in which a deletion, addition or substitution has been introduced, to alter accordingly, for example, to affect functionally, the endogenous gene. Preferably, this endogenous gene is a gene of C. gl utami cum or E. coli, but it can be a homologue of a related bacterium or even of a yeast or plant source. In one embodiment, the vector is designed so that, with homologous recombination, the endogenous gene is functionally affected (i.e., it no longer encodes a functional protein; also referred to as a "deactivated" vector). Alternatively, the vector can be designed so that, with homologous recombination, the endogenous gene mutates or otherwise alters but still encodes the functional protein (eg, the upstream regulatory region can be altered to alter the expression of the endogenous enzyme of Table 1). In the homologous recombination vector, the altered portion of the endogenous gene is flanked at its 5 'and 3' ends by the additional nucleic acid of the endogenous gene to allow homologous recombination to be present between the exogenous gene carried by the vector and a gene endogenous in the (micro) organism. The additional flanking endogenous nucleic acid is of sufficient length for successful homologous recombination with the endogenous gene. Typically, several kilobases of flanking DNA (at the 5 'and 3' ends) are included in the vector (see for example, Thomas, KR, and Capecchi, MR (1987) Cell 51: 503 for a description of homologous recombination vectors ). The vector is introduced into a microorganism (for example, by electroporation) and cells are selected in which the introduced endogenous gene has recombined in homologous manner with the endogenous enzymes of Table 1, using techniques known in the art. In another embodiment, an endogenous gene for the enzymes of Table 1 in a host cell is affected (eg, by homologous recombination or other genetic means known in the art) so that the expression of its protein product does not occur. In another embodiment, an endogenous or introduced gene of the enzymes of Table 1 in a host cell has been altered by one or more point mutations, deletions, or inversions, but still encodes a functional enzyme. In yet another embodiment, one or more of the regulatory regions (e.g., a promoter, repressor, or inducer) of an endogenous gene for the enzymes of Table 1 in a (micro) organism has been altered (e.g., by deletion). , truncation, inversion, or point mutation) so that the expression of the endogenous gene is modulated. One of ordinary skill in the art will appreciate that host cells that contain more than one of the genes encoding the enzyme in Table 1 and modifications of the proteins can be easily produced using the methods of the invention, and are intended to included in the present invention. Additionally, a gene repression (but also gene overexpression) is also possible by means of specific DNA binding factors, for example factors of the type of zinc finger transcription factors. Additionally, factors that inhibit the target protein itself can be introduced into a cell. The protein binding factors for example may be the aptamers mentioned in the above (Famulok et al., (1999) Curr Top My crobiol Immunol 243, 123-36). As additional factors of protein binding, whose expression in organisms causes a reduction in the amount and / or activity of the enzymes in Table 1, antibodies specific for the enzymes can be considered. The production of monoclonal, polyclonal, or recombinant antibodies specific to the enzymes follows standard protocols (Guide to Protein Purification, Meth, Enzymol, 182, pp. 663-679 (1990), M. P. Deutscher, ed.). Antibody expression is also known from the literature (Fiedler et al., (1997) Immunotechnology 3, 205-216; Maynard and Georgiou (2000) Annu., Rev. Biomed, Eng. 2, 339-76). The techniques mentioned are well known to the person skilled in the art. Therefore, he also knows what dimensions the nucleic acid constructs used for example for antisense methods and what complementarity, homology or identity should have the respective nucleic acid sequences must have. The terms complementarity, homology and identity are known to the person skilled in the art. Within the scope of the present invention, sequence homology and homology, respectively, are generally understood to imply that the nucleic acid sequence or the amino acid sequence, respectively, of a DNA molecule or a protein, respectively, is at least 25%, at least 30%, at least 40%, preferably at least 50%, further preferred at least 60%, also preferably at least 70%, also preferably at least 80%, preferably particular at least 90%, preferred in particular at least 95% and more preferably at least 98% identical to the nucleic acid sequences or amino acid sequences, respectively, of a known DNA or RNA molecule or protein, respectively. In the present, the degree of homology and identity, respectively, refers to the full length of the coding sequence.

The term "complementarity" describes the ability of a nucleic acid molecule to hybridize with another nucleic acid molecule due to the hydrogen bonds between two complementary bases. The person skilled in the art knows that two nucleic acid molecules must not have a complementarity of 100% in order to be able to hybridize with each other. A nucleic acid sequence, which will hybridize with another nucleic acid sequence, is preferred to be at least 30%, at least 40%, at least 50%, at least 60%, preferably at least 70% , particularly preferably at least 80%, also particularly preferably at least 90%, particularly preferred at least 95% and most preferably at least 98 or 100%, respectively, complementary to the other nucleic acid sequence . The nucleic acid molecules are identical if they have identical nucleotides in identical 5 '-3' order. Hybridization of an antisense sequence with an endogenous mRNA sequence typically occurs in vivo under cellular or in vi tro conditions. According to the present invention, the hybridization is carried out in vivo or in vi tro under conditions that are sufficiently astringent to guarantee a specific hybridization. The astringent hybridization conditions in vi tro are known to the person skilled in the art and can be taken from the literature (see for example Sambrook et al., Molecular Cloning, Cold Spring Harbor Press). The term "specific hybridization" refers to the case where a molecule preferentially binds to a certain nucleic acid sequence under stringent conditions, if this nucleic acid sequence is part of a complex mixture for example of DNA or RNA molecules. The term "astringent conditions" therefore refers to conditions, under which a nucleic acid sequence preferentially binds to an objective sequence, but not, or at least to a significantly reduced extent, to other sequences. The astringent conditions depend on the circumstances. The longer sequences hybridize specifically at higher temperatures. In general, the astringent conditions are chosen such that the hybridization temperature lies about 5 ° C below the melting point (Tm) of the specific sequence with a defined ionic strength and a defined pH value. Tm is the temperature (with a defined pH value, a defined ionic strength and a defined concentration of nucleic acids), in which 50% of the molecules, which are complementary to an objective sequence, hybridize with the target sequence. Typically, the astringent conditions comprise salt concentrations between 0.01 and 1.0 M sodium ions (or other salt ions) and a pH value between 7.0 and 8.3. The temperature is at least 30 ° C for short molecules (for example for molecules such that they comprise between 10 and 50 nucleotides). In addition, the astringent conditions may comprise the addition of destabilizing agents such as formamide. Typical hybridization and washing buffers are of the following composition.

Pre-hybridization solution: 0.5% SDS 5x 50 mM SSC NaP04, pH 6.8 0.1% Na 5x pyrophosphate Denhardt's reagent 100 μg / salmon sperm Hybridization solution: Pre-hybridization solution lxlO6 cpm / ml of probe (5-10 min 95 ° C) 20x SSC: 3 M NaCl 0.3 M sodium citrate aj pH 7 with 50x HCl of Denhardt reactive: 5 g of Ficoll 5 g of polyvinylpyrrolidone 5 g of Bovine Serum Albumin aj to 500 ml A. dest. A typical procedure for hybridization is as follows: Optional: wash the hybridization membrane 30 min at lx SSC / 0.1% SDS at 65 ° C Pre-hybridization: at least 2 h at 50-55 ° C Hybridization: overnight at 55-60 ° C Washing: 05 min 2x SSC / 0.1% SDS Hybridization temperature 30 min 2x SSC / 0.1% SDS Hybridization temperature 30 min lx SSC / 0.1% SDS Hybridization temperature 45 min 0.2x SSC / 0.1% SDS 65 ° C 5 min O.lx SSC room temperature The terms "sense" and "antisense" as well as "Antisense orientation" are known to the person skilled in the art. Additionally, the person skilled in the art knows how long the nucleic acid molecules to be used for antisense methods should be, and what homology or complementarity they should have relative to their target sequences. Accordingly, the person skilled in the art also knows how long the nucleic acid molecules should be, which will be used for gene silencing methods. For antisense purposes, complementarity over sequence lengths of 100 nucleotides, 80 nucleotides, 60 nucleotides, 40 nucleotides and 20 nucleotides may suffice. Longer nucleotide lengths will certainly also suffice. A combined application of the methods mentioned in the above is also conceivable. If, according to the present invention, DNA sequences are used, which are operatively linked in 5'-3 'orientation to an active promoter in the organism, vectors can generally be constructed which, after transfer to the cells of the organism, allow the overexpression of the coding sequence or cause the suppression or competition and blocking of endogenous nucleic acid sequences and the expressed proteins thereof, respectively. The activity of a particular enzyme can also be reduced by over-expressing a non-functional mutant thereof in the organism. In this way, a non-functional mutant that is not able to catalyze the reaction in question, but is capable of binding, for example, the substrate or co-factor, by means of overexpression, can competitively overcome the enzyme endogenous and therefore inhibit the reaction. Additional methods for reducing the amount and / or activity of an enzyme in a host cell are well known to the person skilled in the art.

Vectors and Host Cells Another aspect of the invention pertains to vectors, preferably expression vectors, which contain a nucleic acid encoding the enzymes of the invention.

Table 1 (or portions thereof) or combinations thereof. As used herein, the term "vector" refers to a nucleic acid molecule capable of transporting another nucleic acid to which it has been linked. One type of vector is a "plasmid," which refers to a loop of double-stranded DNA in which additional segments of DNA can be ligated. Another type of vector is a viral vector, where additional segments of DNA can be linked in the viral genome. Certain vectors are capable of autonomous replication in a host cell into which they are introduced (eg, bacterial vectors having a bacterial origin of replication and mammalian episomal vectors).

Other vectors (e.g., non-episomal mammalian vectors) are integrated into the genome of a host cell upon introduction into the host cell, and are consequently replicated together with the host genome. Moreover, certain vectors are capable of directing the expression of genes to which they bind in an operative way. Such vectors are referred to herein as "expression vectors". In general, expression vectors of utility in recombinant DNA techniques are often in the form of plasmids. In the present specification, "plasmid" and "vector" can be used interchangeably since the plasmid is the most commonly used form of the vector. However, the invention is intended to include such other forms of expression vectors, such as viral vectors (e.g., retroviruses, adenoviruses and adeno-associated viruses replication defective), which serve equivalent functions. The recombinant expression vectors of the invention may comprise a nucleic acid encoding the enzymes of Table 1 in a form suitable for the expression of the respective nucleic acid in a host cell, which means that the recombinant expression vectors include one or more regulatory sequences, selected on the basis of the host cells that will be used for expression, which are operatively linked to the nucleic acid sequence to be expressed. Within a recombinant expression vector, "operably linked" is intended to imply that the nucleotide sequence of interest binds to the regulatory sequence (s) in a particular manner that allows for the expression of the nucleotide sequence (e.g. transcription / translation system in vitro or in a host cell when the vector is introduced into the host cell). The term "regulatory sequence" is intended to include promoters, repressor binding sites, activator binding sites, enhancers, and other expression control elements (eg, terminators, polyadenylation signals, or other elements of secondary structure of mRNA) . Such regulatory sequences are described, for example, in Goeddel; Gene Expression Technology: Methods in Enzymology 185, Academic Press, San Diego, CA (1990). Regulatory sequences include those that direct the constitutive expression of a nucleotide sequence in many types of host cells and those that direct the expression of the nucleotide sequence only in certain host cells. Preferred regulatory sequences are, for example, promoters such as cos-, tac-, trp-, tet-, trp-, tet-, lpp-, lac-, lpp-lac-, laclq-, T7-, T5-, T3 -, gal-, trc-, ara-, SP6-, arny, SP02, e-Pp- or PL, which are preferably used in bacteria. Additional regulatory sequences are, for example, promoters of yeasts and fungi, such as ADCl, MFa, AC, P-60, CYCl, GAPDH, TEF, rp28, ADH, plant promoters such as CaMV / 35S, SSU, OCS, lib4 , usp, STLS1, B33, us or ubiquitin or phaseolin promoters. It is also possible to use artificial promoters. It will be appreciated by one of ordinary skill in the art that the design of the expression vector may depend on factors such as the choice of the host cell to be transformed, the level of expression of the desired protein, etc. The expression vectors of the invention can be introduced into host cells to thereby produce proteins or peptides, including proteins or fusion peptides, encoded by the nucleic acids encoding the enzymes of Table 1. The recombinant expression vectors of the invention can designed for the expression of the enzymes in Table 1 in prokaryotic or eukaryotic cells. For example, genes for the enzymes of Table 1 can be expressed in bacterial cells such as C. glutamicum and E. coli, insect cells (using baculovirus expression vectors), yeast cells and other fungal cells (see Romanos, MA et al. (1992), Yeast 8: 423-488, van den Hondel, CA MJ. (1991) in: More Gene Manipulations in Fungi, JW Bennet &LL Lasure, eds., Pp. 396-428: Academic Press: San Diego, and van den Hondel, CAMJJ &Punt, PJ (1991) in: Applied Molecular Genetics of Fungi, Peberdy, JF et al., Eds., Pp. 1-28, Cambridge University Press: Cambridge), algae and multicellular plant cells (see Schmidt, R. and Willmitzer, L. (1988) Plan t Cell Rep .: 583-586). Suitable host cells are further discussed in Goeddel, Gene Expression Technology: Methods in Enzymology 185, Academic Press, San Diego, CA (1990). Alternatively, the recombinant expression vector can be transcribed and translated in vitro, for example using promoter regulatory sequences of T7 and T7 polymerase. The expression of proteins in prokaryotes is most often carried out with vectors containing constitutive or inducible promoters that direct the expression of fusion or non-fusion proteins. The fusion vectors add a number of amino acids to a protein encoded therein, usually to the amino terminus of the recombinant protein but also to the C-terminus or fuse them within suitable regions in the proteins. Such fusion vectors typically serve three purposes: 1) to increase the expression of the recombinant protein; 2) to increase the solubility of the recombinant protein; and 3) to aid in the purification of the recombinant protein by acting as a ligand in affinity purification. Often, in fusion expression vectors, a proteolytic cleavage site is introduced into the junction of the fusion portion and the recombinant protein to allow separation of the recombinant protein from the fusion portion subsequent to purification of the protein. of fusion. Such enzymes, and their known recognition sequences, include Factor Xa, thrombin and enterokinase. Typical fusion expression vectors include pGEX (Pharmacia Biotech Inc. Smith, D. B. and Johnson, K. (1988) Gene 67: 31-40), pMAL (New England Biolabs, Beverly, MA) and pRIT5 (Pharmacia, Piscataway, NJ) which fuse glutathione S-transferase (GST), maltose binding protein E, or protein A, respectively. Examples of suitable expression vectors in E. coli without inducible fusion include pTrc (Amann et al., (1988) Gene 69: 301-315), pLG338, pACYC184, pBR322, pUC18, pUC19, pKC30, pRep4, pHSl, pHS2, pPLc236, pMBL24, pLG200, pUR290, pIN -III113-Bl, egtll, pBdCl, and pET lid (Studier et al., Gen Expression Technology: Methods in Enzymology 185, Academic Press, San Diego, California (1990) 60-89, and Pouwels et al., Eds. 1985) Cloning Vectors, Elsevier: New York IBSN 0 444 904018). Expression of the target gene from the pTrc vector uses transcription of the host RNA polymerase from a trp-lac hybrid fusion promoter. The expression of the target gene from the pET lid vector uses the transcription of a T7 gnlO-lac fusion promoter mediated by a co-expressed viral RNA polymerase (T7gnl). This viral polymerase is supplied by the BL21 (DE3) or HMS 174 (DE3) host strains of a resident X-profane harboring a T7gnl gene under the transcriptional control of the lacUV 5 promoter. For the transformation of other strains of bacteria, appropriate vectors can be selected For example, plasmids pIJIOl, pIJ364, pIJ702 and pIJ361 are known to be useful for transforming Streptomyces, whereas plasmids pUBUO, pC194, or pBD214 are suitable for the transformation of Bacillus species. Several plasmids for use in the transfer of genetic information in Corynebacterium include pHM1519, pBLl, pSA77, or pAJ667 (Pouwels et al., Eds. (1985) Cloning Vectors.Elsevier: New York IBSN 0 444 904018). One strategy to maximize the expression of recombinant proteins is to express the protein in a host bacterium with an impaired ability to proteolytically cleave the recombinant protein (Gottesman, S., Gen Expression Technology: Methods in Enzymology 185, Academic Press, San Diego, California (1990) 119-128). Another strategy is to alter the nucleic acid sequence of the nucleic acid to be inserted into an expression vector so that the individual codons for each amino acid are those used preferentially in the bacterium chosen for expression, such as C. gl utami cum (Wada et al. (1992) Nucleic Acids Res. 20: 2111-2118). Such alteration of nucleic acid sequences of the invention can be carried out by standard techniques of DNA synthesis. Examples of suitable shuttle vectors of C. gl utami cum and E coli can be found in Eikmanns et al (Gene. (1991) 102, 93-8). In another embodiment, the protein expression vector is an expression vector in yeast. Examples of vectors for expression in the yeast S. cerevisiae include pYepSecl (Baldari, et al., (1987) Embo J. 6: 229-234), 2i, pAG-1, Yep6, Yepl3, pEMBLYe23, pMFa (Kurjan and Herskowitz, (1982) Cell 30: 933-943), pJRY88 (Schultz et al., (1987) Gene 54: 113-123), and pYES2 (Invitrogen Corporation, San Diego, CA). Vectors and methods for construction of vectors suitable for use in other fungi, such as filamentous fungi, include those detailed in: van den Hondel, C.A.M. J. J. & Punt, P. J. (1991) in: Applied Molecular Genetics of Fungi, J.F. Peberdy, et al., Eds., P. 1-28, Cambridge University Press: Cambridge, and Pouwels et al., Eds. (1985) Cloning Vectors. Elsevier: New York (IBSN 0444 904018). For the purposes of the present invention, an operative link is understood as the sequential disposition of • promoter, coding sequence, terminator and, optionally, additional regulatory elements in such a way that each of the regulatory elements can fulfill its function, in accordance with its determination, when the coding sequence is expressed. In another embodiment, the proteins of Table 1 may be expressed in unicellular plant cells (such as algae) or in plant cells of higher plants (e.g., spermatophytes, such as crop plants). Examples of plant expression vectors include those detailed in: Becker, D., Kemper, E., Schell, J. and Masterson, R. (1992) Plan t Mol. Biol. 20: 1195-1197; and Bevan, M. W. (1984) Nucí. Acid Res. 12: 8711-8721, and include pLGV23, pGHlac +, pBIN19, pAK2004, and pDH51 (Pouwels et al., Eds. (1985) Cloning Vectors.Elsevier: New York IBSN 0 444 904018). For other expression systems suitable for prokaryotic and eukaryotic cells see chapters 16 and 17 of Sambrook, J. et al. Molecular Cloning: A Laboratory Manual. 3rd ed. , Cold Spring Harbor Laboratory, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, 2003. For the purposes of the present invention, an operative link is understood as the sequential arrangement of promoter, coding sequence, terminator and, optionally, regulatory elements. additional in such a way that each of the regulatory elements can fulfill its function, according to its determination, when the coding sequence is expressed.

In another embodiment, the recombinant expression vector in mammals is capable of directing the expression of the nucleic acid preferentially in a particular cell type, for example in plant cells (for example, tissue-specific regulatory elements are used to express the nucleic acid ). The tissue-specific regulatory elements are known in the art. Another aspect of the invention pertains to host organisms or cells in which a recombinant expression vector of the invention has been introduced. The terms "host cell" and "recombinant host cell" are used interchangeably herein. It is understood that such terms refer not only to the particular target cell but also to the progeny or potential progeny of such a cell. Since certain modifications may occur in successive generations due to mutation or environmental influences, such progeny may, in fact, not be identical to the progenitor cell, but are still included within the scope of the term as used herein. A host cell can be any prokaryotic or eukaryotic cell. For example, an enzyme of Table 1 can be expressed in bacterial cells such as C. gl utamicum or E. coli, insect cells, yeast or plants. Those of ordinary skill in the art know other suitable host cells.

The vector DNA can be introduced into prokaryotic or eukaryotic cells by conventional transformation or transfection techniques. As used herein, the terms "transformation" and "transfection", "conjugation" and "transduction" are intended to refer to a variety of techniques recognized in the art for introducing foreign nucleic acid (e.g., linear DNA or RNA (eg. example, a linearized vector or a single gene construct without a vector) or nucleic acid in the form of a vector (eg, a plasmid, phage, phasmid, phagemid, transposon or other DNA) in a host cell, including co-precipitation with calcium phosphate or calcium chloride, DEAE-dextran-mediated transfection, lipofection, natural competence, chemical mediated transfer, or electroporation Suitable methods for transforming or transfecting host cells can be found in Sambrook, et al. (Molecular Cloning: A Laboratory Manual, 3rd edition, Cold Spring Harbor Laboratory, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, 2003), and other laboratory manuals. To identify and select these members, a gene encoding a selection marker (e.g., antibiotic resistance) is generally introduced into the host cells together with the gene of interest. Preferred selection markers include those that confer resistance to drugs, such as G418, hygromycin and methotrexate. The nucleic acid encoding a selection marker can be introduced into a host cell in the same vector as that encoding the enzymes in Table 1 or can be introduced into a separate vector. Cells stably transfected with the introduced nucleic acid can be identified by drug selection (for example, cells that have incorporated the selection marker gene will survive, while the other cells will die). In another embodiment, recombinant microorganisms may be produced that contain selected systems that allow regulated expression of the introduced gene. For example, the inclusion of a gene of Table 1 in a vector by placing it under the control of the lac operon allows the expression of the gene only in the presence of IPTG. Such regulatory systems are well known in the art. In one embodiment, the method comprises culturing the organisms of the invention (in which a recombinant expression vector encoding for example an enzyme of Table 1 has been introduced, or in whose genome a gene encoding an enzyme type has been introduced) wild or altered) in a medium suitable for production of methionine. In another modality, the method further comprises isolating methionine from the medium or the host cell. It has been stipulated in the foregoing that in order to modulate the metabolic flux of an organism, the amount and / or activity of the enzymes in Table 1 that catalyze a metabolic network reaction may be increased or reduced. However, in order to modify the metabolic flux of an organism to produce an organism that is more efficient in methionine synthesis, changing the amount and / or activity of an enzyme is not limited to the enzymes listed in Table 1. Any enzyme that is homologous to the enzymes of Table 1 and performs the same function in another organism can be perfectly suitable to modulate the amount and / or activity in order to influence the metabolic flow by means of over-expression. The definitions for homology and identity have been given in the above. In the following table, examples of homologs are given to some of the enzymes Rl to R61 of Table 1 which can be used for the purposes of the present invention for example by over-expressing them in C. glutami cum or E. coli in order to increase the amount and / or activity of the respective enzymes: Table 2 Function Name Sequence numbers of accession products of gene product access related / alternative proteins, proteins with conserved activity (EC number) recovery system CAA52144.1 YP_079782.1; NP_980596.1; YP _021091.1 Glycine cleavage,; AA11139.1; YP_148276.1, YP_175989.1; Glycine protein T from B? B6565.1; NP_464875.1; NP_470723.1; (R71 / R72) amino methyl- YP_013965.1; ZP_00231385.1; ZP_005385 trapsferase 77.1; NP_692823.1; NP_764775.1; YPJ 88 676.1; NP_372059.1; C? G43268.l; YP_253 296.1; YP_186433.1, YP_041008.1; NP_62 1985.1; ZP_00560257.1; AAU84894.1; ZP_ 00574838.1; YP_075748.1; NP_143816.1; C AB50682.1; NP_662997.1; NP_972230.1; A? 82124.1, ZP_OO590815.1; ZP_00528533 .1; B? D85568.1; NP_228029.1; ZP_005111 04.1; ZP_005 1209.1; ZP_00532593.1; ZP_ 00525162.1; ZP_00355911.1; YP_112880.1; ZP_00399764.1; YP_143792.1; CAD7544 8.1; YP_004126.1; ZP_00535960.1; ZP_003 34892.1; C? G35027.1; C? F23008.1; YP_0 10643.1; NP_951437.1; NP_840694.1; EAN 28088.1; YP_125490.1; YP_094168.1;? AO 91208.1, YP_122478.1; NP_110817.1, NP_ 214308.1; CAC12478.1; ZP_00602311.1; Y P_256007.1; E? M94548.1; B? B66249.1; NP 342409.1; YP_023949.1; ZP_00054699.1; ?? K25314.1; ZP_00270640.1; YP_16945 5.1; NP_148400.1; ZP_00577149.1; ZP_003 75766.1; ZP_00303628.1; NP_280387.1; A? V46419.1; E? O04791.1; C? C32302.1; C? I36361.1;? Z12256. 1; NP_013914.1; YP _034020.1; BAC74698.1; Q827D7; ZP_003 96527.1; EAK92665.1; EAK92694.1; ZP_0 0292858.1; ZP_00625542.1; NP_298674.1; EA014274.1; NP_883104.1; NP_887405.1; ZP_0O038971.2; NP_778843.1; ZP_003276 36.1; EAA72140.1; NP_879086.1; NP_1025 91.1; CAJ04455.1; C? G61762.1; NP_71641 2.1; C AG88846.1; ZP_00582521.1; CAD 17 083.1;? AM36086.1; YP_191522.1; EAM7 5690.1; CAH00726.1; CAF26479.1; AAF11 360.1; YP_202186.1; ZP_00308932.1; EAA 47849.1; ZP_00278041.1; CA A81076.1; CA A91000.1, CAA85353.1; EAM85167.1; YP _071681.1; ZP_00638529.1; CAB 16911.1; EAN06353.1; BAD35509.1; EAL84525.1; YP_118701.1; AAQ24377.1; YP_223286.1; NP_541539.1; ZP_00634544.1; CAB16916. 1 BAD82264.1;? AN47559.1; NP_772393. YP_000299.1; CAB16918.1; CAD52982. XP_314216.2; NP_930808.1; AAD56281. AAQ66378.1; AA076254.1; AAN33907. CAG73659.1; XP_219785.3; ZP_001116 07.1; ZP_00585062.1; AAL21928.1; AAH4 2245.1; AAX66900.1; AAS46734.1; NP_94 9187.1; NP_989653.1; AAC31228.1;; AAA 63798.1; ZP_00004510.1; CAA42443.1; ZP _00555139.1; YP_132995.1; NP_251135.1; NP_636487.1; ZP_00213263.1; ZP_001401 78.2; ZP_00516950.1; AAB82711.1; AAZ2 7773.1; EAA61388.1; ZP_00167208.2; EA N04068.1; AAO07159.1; NP_936747.1; ZP _00499479.1; NP_806663.1; AAW42121.1; CAG83849.1; NP_923192.1; NP_441838.1; ZP_00593912.1; CAA52146.1; BAA14286. 1; Q8XD33; AAA69071.1; EAM28058.1; ZP _00494650.1; ZP_00489316.1; ZP_004706 29.1; YP_152074.1; ?? M14125.1; ?? M91 322.1; ZP_00569907.1; ZP_00459442.1; NP _755358.1; ZP_00436271.1; NP_613061.1; CAE59244.1; BAC38022.1; CAA91099.1; ZP_00264533.1; AAC46780.1; AAG58030. 1; YP_261725.1; AAL24244.1; NP_708666. 1; CAH07776.1; Q8YNF9; YP_099306.1; Z P_00474391.1; AAL57651.1; YP_055456.1; NP_682393.1; NP_893785.1; YP_172756. 1; C? H74116.1; ZP_00622638.1; YP_23 1 88.1; NP_791106.1; NP_967658.1; EAN856 56.1; ZP_00379711.1; XP_520482.1; A AN6 6613.1; ZP_00162707.1; ZP_00417692.1; Y P_264083.1; AAQ61092.1; NP_216348.1; N P_8555l5.1; CAE76410.1; AAO39460.1; C AE22343.1; NP_800311.1; YP_206661.1; Z P_00506636.1; ZP_00417032.1; YP_16489 0.1; XP_760322.1; YP_266091.1; YP_1564 73.1; C? C46126.; ZP_00244924.1; E? N9 1112.1; XP_637330.1; NP_960479.1; YP_1 60522.1; ZP_00317484.1; ZP_00412767.1; NP_253900.1; ?? Q00873.1; NP_532152.1; ZP_00151464.1; E? O09970.1; ZP_006310 36.1; ZP_00141690.2; NP_302381.1; CAGO 8109.1; CAE08889.1; YP_263017.1; AAN7 0757.1; ZP_ 0264788.1; XP_538655.1; AA Z27305.1; AAN17423.1;; AAO38610.1; CA B85154.1; AAW89976.1; AA044232.1; NP _789087.1; ZP_00496567.1; H57478.1; AAS16361.1; AAL33595.1; ZP_00048418. 1; CAA72255.1; XP_598207.1; BAD82265. 1; ?? K26613.1; AAM93931.1; XP_517277 .1; CAB50683.1; NP_143817.1; AAL82123. 1; BAD82266.1; EAN80863.1; BAD85569. 1; ZP_00574837.1; BAB26854.1; ZP_00560 256.1; AAU84893.1; AAD 33990.1; YP_175 990.1;; ZP_00525161.1; NP_148401.1; ZP_00589915.1; YP_079783 .1; ZP_00233535.1; ZP_00577150.1; NP_22 8028.1; ZP_00238490.1; NP_980597.1; YP_ 021092.1; AAP11140.1; YP_038289.1; BA B06534.1; ZP_00399763.1; YP_148277.1; XP_606427.1; AAL04442.1; YP_075749.1; ZP_00527784.1; NP_692824.1; ZP_003348 95.1; C? D75449.1; NP_840693.1; ZP_0059 1425.1; CAF23007.1; ZP_00054162.1; NP_ 621986.1; CAC12479.1; NP_662508.1; ZP_00512041.1; AAV46420.1; ZP_00375765.1; NP_110816.1; NP_280388.1; BAB66248.1; ZP_00538578.1; NP_951436.1; ZP_00367 500.1; YP_169454.1; AAX77943.1; YP_094 170.1; YP_122480.1; YP_125492.1;; ZP_00 270641.1; YP_178313.1; XP_473181.1; ZP_ 00535961.1; NP_372060.1; ZP_00602310.1; YP_255986.1; CAE05443.2; CAB72709.1; CAG43269.1; NP_342410.1; BAB95353.1; AAA92036.1; YP_186434.1; YP_041009.1; AAM63785.1; XP_584346.1; AAM51302.1; AAF99780.1; ZP_00563258.1; EAN09988. 1; NP_148290.1; YP_253295.1; ZP_006272 78.1; YP_256008.1; AA091209.1; XP_6552 57.1; ZP_00303629.1; CAC11669.1; NP_39 4004.1; E? M93962.1; CAB57769.1; C? B5 0124.1; ZP_00520055.1; ZP_00369709.1; E AN28089 .1; CAC 12627.1; AAB85605.1; Q 9HI38; AAH26135.1; EAM24571.1; AAC0 3768.1; AAP98645.1; CAJ18422.1; NP_213 757.1; CAA05909.1; NP_142859.1; BAD32 415.1; BAC31437.1; NP_947805.1; YP_080 623.1; CAH93356.1; ZP_00600159.1; NP_7 75139.1; ZP_00400734.1; CAA09590.2; XP _321361.2;? L81283.1; BAB81491.1; ZP _00506445.1; ZP_00533640.1; YP_160688. 1; NP_764776.1; NP_967201.1; AAH52991. 1; AA038289.1; NP_837365.1; XP_521504. 1; NP_371248.1; NP_753970.1; CAG42465. 1; AAC74750.1; AAU93791.1; Q7ADI4; 1 K MK; 1 JF9; BAA86566.1; YP_170184.1; YP_ 165179.1; NP_624289.1; AAX77932.1; CA D14721.1; BAD84717.1; ZP_00048472.2; A? F73526.1; XP_641773.1; ZP_00570672; .1; ZP_00400089.1; NP_391148.1; NP_001007 947.1; CAE08806.1; CAB50330.1; YP_192 068.1; ZP_00152262.1; ZP_00502920.1; YP _096188.1; NP_796975.1; YP_254084.1; Y P_l 24440.1; BAD86003.1; CAH04407.1; B AB07188.1; Q9K7A0; XP_313472.2; NP_1 03175.1; AAF63783.1; AAN58019.1; YP_1 27437.1; AAM62682.1; CAG90500.1; AAB 85857.1; ZP_00387883.1; XP_562557.1; O2 7433; ZP_00170920.2; C ?? 63066.1; E? N0 8419.1; CAB73027.1; ZP_00335679.1; AA B85866. 1; ZP_00129032.1; YP_178855.1; YP_145876.1; AA V94639.1; NP_623991.1; YP_04O300.1; YP_040204.1; YP_080545.1 ; YP_253952.1; ZP_00638523.1; NP_37136 8.1; YP_255707.1; AAU38404.1; NP_08567 8.1; AAK68403.2; BAB66416.1; XP_62494 4.1; ZP_00135005.2; ZP_00595047.1; NP_9 50100.1; ZP_00458730.1; Q75173.1; ? P99129.1; YP_004062.1; XP_601722.1; C AC45089.1; CAA07007.1; EAL01528.1; XP _694140.1; NP_770978.1; A? C49935.1; EA A21518.1 recovery system Q8FE66. NP 391159: Glycine cleavage: YP_152075.1; NP_806664.1; NP_755359.1 Glycine protein H; Q8FE66;? AX66901.1; ZP_00585063.1; N (R71 R72) P_716411.1; ZP_00582520.1; NP_930809.1; YP_071682.1; ZP_00638530.1; CAG7365 8.1; YP_156474.1; ZP_00634545.1; YP_131 233.1, YP_268018.1; ZP_00417033.1; AAN 70758.1; ZP_00141691.2; NP_253901.1;? AZI 8651.1; E? 018275.1; ZP_00264789.1; ZP_00474390.1; ZP_00499480.1; ZP_0045 9441.1; EAM28059.1; ZP_00318113.1; AA Q61093.1; NP_790167.1; AAO91210.1; CA H37374.1; YP_169453.1; AAW49868.1; NP _840692.1; ? AO07160.1; YP_263019.1; Q9 K0L7; YP_160523.1; NP_800312.1; ZP_00 334896.1; ZP_00278042.1; YP_233358.1; A AW90049.1; ZP_00213264.1; AAF96187.1; CAB84042.1; AAM37905. 1; ZP_0065847 8.1; YP_112591.1; YP_132994.1; YP_2004 34.1; ZP_00593911.1; YP_206660.; ZP_00 167207.1; XP_464281.1; NP_638224.1; E? M75684.1; ZP_00574836.1; ZP_00151463. 1; YP_270507.1; CAD17082.1; BAD45416. 1; YP_004124.1; YP_143790.1; YP_075750 .1; ?? F11361.1; NP_465948.1; C? G35029. 1; NP_960473.1; ZP_00568923.1; EAN0635 2.1; ZP_00506635.1; NP_532153.1; YP_223 285.1; NP_541538.1; YP_014985.1; AAO63 775.1; CAA20174.1; ZP_00399762.1; YP_0 10645.1; NP_471849.1; AA077626.1; YP_094171.1; NP_693309.1; BAC70485.1; NP_2 28027.1; YP_253966.1; 1ZK0; YP_148857. 1; YP_046528.1; YP_040289.1; ZP_001311 07.1; YP_176481.1; NP_297474.1; Q9PGW 7; ZP_00355907.1; ZP_00651773.1; NP_77 8393.1; YP_021882.1; NP_855509.1; CAF2 6480.1; CAG42548.1; ZP_00389942.1; NP_ 981423.1; AAP11863.1; NP_302386.1; NP_ 764151.1; YP_266090.1; ZP_00237746.1; Z P_00244923.1; ZP_00396528.1; YP_18807 7.1; CAC46127.1; ZP_00375764.1; ZP_005 60255.1; ZP_00270642.1; BAB07203.1; EA N28788.1; NP_216342.1; C? H09835.1; NP _251136.1; YP_185749.1; AAT49691.1; ZP _00264532.1; NP_102590.1; YP_034021.1; NP 879085.1; AAP54618.1; ZP_00625541. 1; ZP_00525160.1 AAV46421.1; ZP_00412 765.1; C AH02746.1; CAA81075.1; CAB 16 912.1; CAB16710.1; CAA85761.1; ZP_006 22637. l CAB 16914.; CAA85759.1; NP_1 43205.1; YP_101635.1; ZP_00380048.1; ZP _00004511.1; ZP_00414055.1; ?? C61829. I; ?? L81616.1; YP_261724.1; C? B49742. 1; AAN66614.1; Q9V0G 1; YP_080558.1; B? D84339.1; AAK25316.1; NP_949186.1; C AJ13836.1;? AQ67414.1; 1DXM; CAA857 68.1; CAA85757.1; P93255; CAJ13723.1; CA? 85755.1; ZP_00292299.1; ZP_00308328 .1; NP_772392.1; CAI36363.1; NP_147622. 1; NP_391159.1; ZP_00577152.1; CAA857 56.1; AAQ66080.1; AAV94183.1; CAA887 34.1; ZP_00547429.1; NP_621789.1; CA? 8 5760.1; AAR37471.1; ZP_00555140.1; AA W47059.1; ZP_00303630.1; NP_621987.1; AAG48828.1; CAG86839.1; YP_191521.1; YP_055457.1; ZP_00631037.1; YP_118696 .1; CAA85767.1; XP_637044.1; A? M64413 .1; CAC19751.1; ZP_00379709.1; NP_2803 89.1; CAF23006.1; EA025275.1; AAU8489 CAF92157.1, CAF92157.1, EAN04067.1; AAH91548 .1; YP_164889.1; NP_213756.1; C? G62852 .1; A? W49010.1; CAA94317.1; ZP_005277 86.1; YP_234189.1; C? D52976. 1; NP_9676 50.1; C? E66592.1; E ?? 66192.1; AAW277 08.1; ZP_00565058.1; XP_536768.1; BAB6 6246.1; CAA95820.1; AAL68248.1;? AH1 4745.1; NP_791107.1; EAL90537.1; XP_57 9628.1; XP_316586.2; AAP88829.1; NP_00 4474.2; NP_080848.1; NP_951435.1; XP_5 23434.1; C? A85754.1; Q9N121;? AS5984 8.1; YP_172757.1; ZP_00534758.1; ZP_005 91423.1; ZP_00512042. 1; ZP_00661686.1; NP_883103.1; ZP_00054163.1; NP_598282 .1; AAW31875.1; XP_756407.1; AAH7621 2.1; CAF99616.1; CAA94316.1; NP_95306 7.1; EAA72139.1; NP_110807.1; AAS5231 5.1; ZP_00575020.1; P20821; NP_662509.1; CAG33353.1; CAE63163.1; ZP_00589916 .1; XP_217678.1; YP_256010.1; XP_58283 5.1; C? C12487.1; NP_394822.1; ZP_00535 962.1; NP_001004372.1; EAL29812.1; CA B05472.1; XP_615385.1; EAL03567.1; O22 535; AAH82740.1; AAX07637.1; NP_9264 77.1 EAM93557.1; XP_584988.1; AAM92 707.1; ZP_00515529.1; ZP_00530979.1; A AH81062.1; ZP_00111606.1; NP_682468.1; Q8Dffi2; EAA77334.1; AAN47560.1; AAL 33596.1; ZP_00162706.2; NP_342412.1; Q8 G4Z7; ZP_00120558.2; ZP_00136571.1; NP _972232.1; CAE 18120.1; Q8YNF8; XP_604 979.1; CAE08890.1 CAE76092.1; YP_023 402.1; NP_009355.2; P39726; CAD75450.1; NP_440920.1; CAE47935.1; XP_498178.1; NP_893786.1; ZP_00327635.1; EAN76953. 1; ZP_00140179.2; EAN99694.1; AAZ1469 6.1; CAE22344.1; EAN83079.1; B AB26349 .1; YP_169819.1; AAX78078.1; AAQ00874 .1; ?? C36844.1; C? G78944.1; XP_694123. 1; ZP_00050263.1; ?? 044734.1; XP_4141 65.1; ZP_00654389.1; ZP_00575538.1; XP_518701.1; BAB66989.1; NP_342534.1; XP_ 583383.1; NP_213643.1; YP_255059.1; NP _214139.1; NP_213280. 1; ZP_00540302.1; YP_055788.1; AAP05107.1; AAF39393.1; ZP_00399135.1; NP_701199.1; CAD75020 .1; XP_343995.2; AAP98380.1; NP_300490 .1; XP_635521.1; YP_219766.1; XP_63705 9.1; ?? V71 155.1; XP_739104.1; E ?? 1807 6.1; NP_219787.1; ZP_00400682.1; NP_967 270.1; ZP_00384691.1; ZP_00401280.1; NP _816146.1; ZP_00526092.1; NP_213714.1; YP_255058.1; NP_785822.1; BAB66988.1; NP_253484.1; ZP_00141248.1; ZP_003991 37.1; XP_676766.1; EAO22790.1; NP_3425 72.1; AAF07900.1; ZP_00152140.2; XP_51 8003.1; AAA23866.1; AAP06383.1; A? M9 9940.1; ZP_00523572.1;? H09065.1; YP_ 039780.1; NP_735539.1; NP_326272.1; YP _252181 .1; NP_802319.1; XP_637062.1; A? K70873.1; ZP_00523574.1; NP_784119.1; XP_356748.3; YP_115837.1; XP_520481. 1; BAB94166.1; XP_527720.1; NP_975513. 1; ZP_00511802.1; ZP_00660576.1; XP_59 8843.1; ZP_00335312.1; ZP_00501149.1; Z P_00496208.1; ZP_00488962.1; ZP_00481 092.1; ZP_00470769.1; YP_112037.1; ZP_0 0315286.1; ZP_00591859.1; NP_622848.l; CAB59889.1; ZP_00566435.1; ZP_006337 79.1; AAG13505.2; CAB16915.1; ZP_0058 5787.1; ZP_00531610.1; AAW51218.1; YP _263215.1; AAK22373.1; AAT58044.1; EA A20319.1; NP_842410. 1; EAN32517.1; NP _532021.1; AAH75478.1; NP_533976.1; ZP _00385854.1; AAL04441.1; AAK89915.1; BAD16654.1; NP_216731.1; NP_770578.1; ZP 00547768.1; NP_522769.1; ZP_003830 64.1; ZP_00322497.1; YP_022846.1; NP_96 1247.1; YP_238125.1; ZP_00412124.1; XP_ 475165.1; XP_470945.1; NP_910410.1; NP _635909.1; CAE01575.2; XP_419793.1; ZP _00170555.1; NP_217017.1;; ZP_00556997 .1; AAP99073.1; NP_297341.1; AAX73221. 1; NP_214109.1; NP_440434.1; CAG87711. 1; ZP_00051435.1; BAB80778.1; XP_39338; 9.2; ZP_00651791.1; ZP_00269419.1; ZP_0 0423019.1; YP_170419.1; NP_001006383. 1; XP_482561.1; NP_778293.1; CAB03400. 1; CAH04871.1; XP_65512J1; AAF61288. 1; ZP_00356683.1; ZP_00565878.1; CAJ01 708.1; XP_758384.1; AAU38115.1; AAK23 858.1; XP_688912.1; BAB06344. recovery system CAA52144 NP_390336; NP_708668.1; Q8XD32; NP_7 glycine cleavage: 55360.1; 1VLO; YP_152076.1; AAX66902. glycine protein P 1; YP_071683.1; NP_670593.1; NP_930810 (R71 / R72) glycine .1; C? G73657.1; ZP_00585064.1; ZP_0063 dehydrogenase 4546.1; NP_716410.1; ZP_00638531.1; ZP_00582519.1; YP_156475.1; YP_268017.1; Z (decarboxylating) P_00141692.2; ZP_00417034.1; NP_25390 2.1jAAT51348.1; ZP_00318114.1; YP_233 357.1; AAO91211.1; AAM37906.1; ZP_002 64790.1; NP_790166.1; YP_125494.1; YP_ 263020.1; YP_122482.1; YP_094172.1; AA N70759.1; Q5ZZ93; YP_20O433.1; NP_638 225.1; YP_1 12882.1; NP_778394.1; NP_84 0691.1; EAO 18276.1; NP_297476.1; ZP_00 651772.1; YP_l 60524.1; AAQ61094.1; ZP_ 00334897.1; YP_104498.1; ZP_00213265.1; EAM28060.1; NP_887403.1; NP_883102. 1; ZP_00151462.1; ZP_00654390.1; NP_87 9084.1; ZP_00459440.1; ZP_00454903.1; Z P_00278043.1; ZP_00499481.1; EAN28090 .l; CAD1708l.l, Q9K0L8; AAZ18652.1; C? B84041.1; Q9JVP2; ZP_00167206.1; YP_ 169452.1; AAW90051.1; AAW49997.1; ZP _00593910.1; ZP_00419736.1; ZP_005602 54.1; YP_041010.1; YP_186435.1; YP_253 294.1; NP_764777.1; YP_079784.1; ZP_003 25013.1; YP_1725O4.1; BAB06535.1; NP_6 21988.1; NP_390337 .1; NP_464873.1; ZP_0 0233534. 1; ZP_00165293.2; YP_013963.1; NP_470721.1; NP_681534.1; BAB76308.1; ZP_00399761.1; ZP_00517920.1; NP_6928 25.1; Q8CXD9; NP_926476.1; Q7NFJ5; YP _075751.1; NP_228026.1; ZP_00162705.1; YP_148278.1; AAQ00895.1; 1 YX2; ZP_00 355906.1; ZP_00111605.1; AAU84891.1; N P_441988.1; YP_085561.1; ZP_00238491.1; NP_980598.1; ZP_00530333.1; NP_66266 7.1; AAP11141.1; CAE22389.1; CAE08940. 1 EA025274.1; ZP_00574835.1; ZP_00511 544.1; NP_967651.1; YP_004123.1; ZP_005 90889.1; YP_175991.1; ZP_00538579.1; YP _143789.1; ZP_00533333.1; ZP_00307846. 1; ZP_00396529.1; ZP_00660916.1; ZP_005 88126.1; ZP_00525159.1; ZP_00531283.1; NP_893804.1; C? I36362.1; AA079689.1; AAX16385.1; NP_393488.1; CAH07007.1; CAA20175.1; NP_110577.1; ZP_00292300 .1; YP_055458.1; BAB59199.1; BAC70484. 1; EAM94419.1; NP_960885.1; ZP_004127 66.1; YP_117906.1; NP_301653.1; NP_972 233.1; NP_295535.1; AAQ66593.1; NP_216 727.1; P64220; ZP_00646130.1; YP_023281 .1; CAC11159.1; 067441; EAM73669.1; CA F23005.1; ZP_00656447.1; AAN47561.1; Y P_000301.1; CAD75451.1; Q8F935; AAV4 6422.1; Q5 V230; ZP_00631038.1; AAO071 63.1; ZP_00379710.1;? AK25317.1; ZP_00 574284.1; ZP_00549460.1; AAL33597.1; N P_936752.1; XP_473945.1; CA? 81081.1; Z P_00054164.1; ZP_00577153.1; C? B 16917 .1; NP_143049.1; O58888; CAB50008.1; BA D86224.1; AAB38502. 1; YP_164888.1; EA L00308.1; NP_280390.1; YP_206658.1; Q9 HPJ7; CAA52800.1; EAL00186.1; ZP_0013 0510.2; AAP21169.1; NP_800315.1; CAA8 1077.1; CAA94902.1; NP_789581.1; CAA1 0976.1; AA044735.1; XP_756577.1; YP_26 6089.1; CAB 11698.1; ZP_00535963.1; NP_ 949185.1; XP_629708.1; ZP_00270643.1;? AK87256.1; AAL81465.1; YP_261727.1; N P_010302.1; EAN06351.1; YP_010902.1; C AG77727.1; AAB05000.1; ZP_00303631.1; NP_001006021.1; ZP_00625540.1; CAG85 941.1; NP_951434.1; NP_772391.1; CAF93 361.1; CAH02226.1; YP_270503.1; CAF26 481.1; ZP_00601921.1; XP_394029.2; ZP_0 0474388.1; CAC46128.1; AAN66611.1; ZP _00264535.1; NP_251132.1; ZP_00375763. 1; ZP_00140175.2; ZP_00555141.1; YP_23 4187.1; AAT51611.1; ZP_00622636.1; ZP_ 00506634.1; NP_791105.1; AAR21108.1; Z P_00417689.1; EAN86200.1; AAW42395.1; EAL33114.1; B AB66247.1; NP_532154.1; YP_034022.1; NP_102589.1; XP_331926.1 ; EAN85387.1; CAG58515.1;; AAB37080.1; ZP_00244922.1; YP_l 91520.1; CAJ09347. 1; C? J09346.1; C? E64583.1; ?? N33909.1; YP_223284.1; NP_214006.1; B ?? 02967.1; BAA03512.1; AAF52996.1; AAX33383.1; XP_322034.2; NP_541537.1; NP_0010138 36.1; EAA68431.1; NP_001014026.1; XP_6 20786.1; EAA51066.1; EAN04066.1; XP_5 41886.1; YP_256009.1; Q9TSZ7; YP_1329 90.1; NP_990119.1; AAH07546.2; EAA657 91.1; Q9YBA2; EAL9O405.1; CAC41491.1; NP_342411.1; EAN79919.1; XP_517018.1; NP_107651.1; ZP_00004512.2; B ?? 12709 .1; NP_148104.1; AAV94849.1; YP_266710 .1; XP_516459.1; NP_105581.1; ZP_006200 69.1; ZP_00631895.1; AAL 13520.1; YP_04 7137.1; AAF68432 .3; CAC46853.1; XP_54 2460.1; ZP_00460296.1; ZP_00554633.1; Z P_OO282393.1; NP_lO29O9.1; YP_235303. 1; AAQ87218.1; ZP_00213445.1; NP_1060 44.1; ZP_00169723.2; ZP_00500952.1; ZP_ 00489849.1; ZP_00480439.1; ZP_0045023 5.1; ZP_0O436058.1;? AY59105.1; EAM32 228.1;? AW21506.1; C ? D14805.1; ZP_004 10721.1; NP_792264.1; Q46337; NP_52160 9.1; BAD97818.1; 1X31; NP_534554.1; ZP_ 00602139.1; AA 1 489.1; YP_266475.1; E AA22341.1; XP_318114.2; CAC47432.1; Z P_00004192.1; NP_534790.1;? AL52901.1; ZP_00565350.1; ZP_00657316.1; YP_134 767.1; ZP_00379371.1; YP_134764.1; XP_7 42683.1; A? N29180.1; ZP_00602144.1; C? G35030.1; EAM31940.1; ZP_00556129.1; CAC49374.1; ZP_00050264.2; ZP_003797 41.1; NP_705537.1; ZP_00471825.1; NP_10 4736.1; ZP_00600293.1; AAY87206.1; YP_ 266690.1; AAL76414.1; AAC31611.1; AA R38319.1; ZP_00658996.1; YP_266673.1; NP_105928.1; ZP_00630198.1; YP_266631 .1; NP_254105.1; NP_107666.1; NP_10428 9.1; NP_102901.1; V96623.1; B AC7466 2.1; AAV95607.1; ZP_00620942.1; ZP_006 45790.1; CAC41486.1; EAN05741.1; AAN 65213.1; YP_269196.1; ZP_00620654.1; ZP _00660136.1; NP_885663.1; NP_436414.1; NP_881143.1; AAM75070.1; NP_572162.2; ZP_00379745.1; CAA39468.1; ZP_002645 73.1; YP_262784.1; ZP_00380782.1; ZP_00 278582.1; AAV94866.1; XP_414684.1; YP_ 237780.1; ZP_00602141.1; YP_165138.1; N P_790307.1; NP_62O8O2.2; NP_534700.1; AAV95690.1; EAL32452.1; AAF21941.1; AAN65956.1; NP_083048.1; AAV94935.1; CAC46854.1; CAC41470.1; AAH24126.1; NP_037523.2; ZP_00565365.1; NP_102887 .1; YP_134760.1; AAQ87217.1; AAH89599 .1; ZP_00554816.1;? AK16482.1; NP_1030 85.1; ZP_00620147.1; NP_102906.1; ZP_00 556188.1; XP_307967.2; ZP_00622120.1; C AC47100.1; NP_106776.1; C? H90377.1; Z P_00554918.1; CAD31640.1; XP_672544.1, AAT81177.1;? AK27867.2; AAD33412.1; NP_107653.1; CAI12276.1; B? D97122.1; Z P_00410737.1; AAD43585.1; NP_103793.1; XP_526883.1; XP_548398.1; C? B63337.2; YP_266661.1; ZP_00521798.1; EAA60688 .1; AAL51865.1; AAV95026.1; EAL26357. 1; ZP_00619907.1; CAE74368.1; NP_10319 0.1; ?? V94915.1; AAG55663.1; XP_39583 1.2; AAK92969.1; NP_446116.1; AAF5779 6.1; AAN7138O.l; ZPJ) OO05411.1; CAE58 942.1; E ?? 70894.1; AAV94873.1; ZP_006 31887.1; ZP_00629673.1; BAB34711.1; ZP _00619923.1; ZP_00622863 .1; EAL85410. 1; AAL76413.1; AAR38318.1; CAD47921. I; AAH03456.1; NP_102854.1; AAH76859. 1; AAL04443.1; AAH68953.1; YP_165137. 1; XP_580581.1; ZP_00050273.2; ZP_0055 6400.1; ZP_00555517.1; AAV96627.1;? A V93943.1; AAH81271.1; AAK87410.1; ZP_ 00556086.1; AAV93879.1; AAH44792.1; X P 27208.1; ZP_00327983.1; ZP_0055421 3.1; NP_532318.1; CAI 12274.1; AAH22388 .1; XP_546052.1; XP_676622.1; AAV93533 .1; YP_265671.1; AAV47350.1; AAV95190, 1; CAD31286.1; ZP_00516133.1; ZP_0062 0892.1;? H55193.1;? AY82706.1; ZP_00 052785.2; AAR38102.1 LpdA system- P0A9P0 NP_706070.2; NP_752095.1; protein cleavage CAA24742.1; AAX64059.1; glycine AAL19118.1; NP_804043.1; (R71 / R72) YP_149503.1; CAG76686.1; YP_069256.1; NP_930833.1; NP_935564.1; ?? O 10051.1; AAF95555.1; ZP_00585786.1; AAK02977.1; NP_798896.1; AAC46405.1; ZP_00122566.1; ZP_00132373.2; YP_131302.1; YP_205561.1; NP_716063.1; ZP_00637900.1; ZP_00633839.1; ZP_00582828.1; AAU37941.1; ZP_00134358.2; ZP_00157402.1; AAX88688.1; NP_439387.1; ZP_00154973.1; AAP96400.1; YP_154852.1; YP_271444.1; ZP_00464633.1; ZP_00451158.1 ZP_00212747.1; ZP_00500723.1 ZP_00486500.1; ZP_00463487.1 ZP_00467577.1; ZP_00423839.1 ZP_00423458.1; ZP_00283805.1 ?? O90013.1; ZP_00595215.1; ZP_00170705.2; NP_879789.1; YP 23783.1; NP_889077.1; YP_126870.1; NP_240038.1; YP_095531.1; C? D15305.1; ?? Q58205.1; NP_883762.1; NP_770362.1; YP 70418.1; CAA61894.1; AAV29309.1; CAB 84783.1; AAF41719.1; ZP_00565931.1; CAA59171.1; CAA54878.1; AAW89295.1; CAA62435.1; ZP_00150164.2; 1BHY; 10JT; CAA61895.1; YP 57096.1; CAA57206.1; AAM38502.1; NP_635936.1; YP_199361.1; NP_660554.1; NP_842161.1; ZP_00507350.1; NP_779995.1; YP_115390.1; ZP_00651360.1; NP_298158.1; AAR38073.1; EAOl 7659.1; ZP_00245305.1; 15 AAR38213.1; AAR38090.1; NP_777818.1; B AC24467.1; NP_891227.1; NP_879460.1; NP_878457.1; C? D71978.1; YP_078853.1; AAK50273.1; AAK50266.1;? AF11916.1; NP_389344.1; ZP_00396676.1; 20 EAO21015.1; CAA37631.1; 1EBD; YP_146914.1; BAB06371.1; YP_085309.1; AAP10890.1; YP_020826.1; YP_175913.1; Q04829; AAN03817.1; NP_692336.1; ?? G17888.1; ZP_00474314.1; NP_764349.1; ??? 99234.1; 1LPF; NP_250278.1; XP_475628.1; YP_253771.1; YP_143499.1; YP_257414.1; AAF34795.3; AAF79529.1; YP_040483.1; YP_005722.1; YP_074243.1; AAN23154.1; AAK50305.1; AAS20O45.1; ZP_00540244.1; 10 EAN07674.1; AAC26053.1; NP_815077.1; NP_908725.1; ZP_00307577.1; AAS47493.1; ? AF34796.1; CAA 11554.1; YP_013676.1; ZP_00317120.1; AAV48381.1; CAB84413.1; AAB30526.1; NP_969527.1; 15 BAB44156.1; NP_464580.1; XP_635122.1; AAF41363.1; ?? K50280.1; ZP_00397330.1; YP 265659.1; NP_470384.1; CAA44729.1; AAW89611.1; 1 DXL; ZP_00401182.1; ZP_00418304.1; NP_792022.1; ZP_00625011.1; AAD53185.1; 3LAD; EAN08634.1; AAH18696.1; CAH93405.1; YP_235092.1; NP_967737.1;; CAJ08862.1; NP_945538.1; NP_763632.1; BAE00452.1; B? D92940.1; NP_000099.1; 1 ZMD; A? B01381.1; NP_999227.1; NP_767089.1; AAS47708.1; AAR21288.1; AAA35764.1; YP_034342.1; EAN90443.1; E AN96941.1; C AA61483.1; AAN69768.1; AAF 12067.1; P31052; NP_105199.1; ZP_00263252.1; AAH62069.1; CAA72132.1; 10 NP_031887.2; CAG58981.1; CAF26798.1; EAN80618.1; ?? N15202.1; CA? 72131.1; ZP_00269527.1; CAD72797.1; ZP_00554136.1; CAD61860.1; AAC53170.1; CAF05589.1; ZP_00622437.1; CAG81278.1; ZP_00284261.1; ZP_00497224.1; 15 EA 93183.1; ZP_00492121.1; NP_533297.1; AAS53883.1; YP_160845.1; AAV93660.1; C? G31211.1; C 49991.1; AAM93255.1; AAK11679.1; ZP_00427535.1; YP_258846.1; A? N30810.1; ZP_00449174.1; 20 CAF92514.1; AAQ91233.1; AAH44432.1; XPJ20877.2; AAH56016.1; YP_222565.1; CAC47627.1; AAA96487.1; ZP_00464142.1; NP_280867.1; YP_067405.1; CAB65609.1; ?? L51327.1; ?? K22329.1; YP_246823.1; NP_105334.1; XP_758608.1; CAH00655.1; NP_772974.1; AAB88282.1; ZP_00211386.1; EAN27796.1; AAN70931.1; EAL29693.1; ZP_00340462.1; ZP_00153792.2; ZP_00579524.1; AAZ17978.1; NP_266215.1; AAN33719.1; 10 ?? D30450.1; ZP_00383074.1; ZP_00597315.1; CAC47514.1; AAF49294.1; YP_223465.1; NP_220840.1; NP_360330.1; E? A26462.1; CAA39235.1; ZP_00578463.1; YP_047424.1; AAM36402.1; BAB03935.1; AAN69982.1; NP_116635.1; 15 ZP_00654346.1; CAG85768.1; 1V59; NP_623271.1; ?? 65618.1; ZP_00305550.1; XP_623438.1; ZP_00007570.1; ZP O0320049.1; AAN75183.1; ZP_00323583.1; YP_200681.1; 1LVL; ZP_00151187.2; AAP03132.1; CAD14973.1; ZP_00630163.1; ZP_00139957.1; ÑP_250940.1; ÑP_636857.1; CAB05249.2; ZP_00166998.2; ?? N75720.1; C 62982.1; ZP_00265019.1; ZP_00384289.1; AAV47687.1; ZP_00303079.1; NP_842316.1; XP_331 183.1; AAV28779.1; A? N48422.1; ZP_00597992.1; AAN75618.1 AAV28746.1; ZP_00267415.1 ZP_00650982.1; AAQ58749.1 NP_298837.1; AAN75159.1; 10 NP_778978.1; ZP_00575798.1; YP_002403.1; ?? B97089.1; ZP_00511405.1; YP_005669.1; EA021998.1; XP_613473.1; ZP_00245417.1; ZP_00210841.1; ZP_00561492.1; YP_259638.1; EA016949.1; NP_785656.1; 15 CAF23812.1; ZP_00055963.2; YP 43553.1; NP_953492.1; CAA63810.1; CAF22875.1; AAV89136.1; ZP_00536790.1; ?? F39644.1; ZP_00621355.1; ZP_00486105.1; ZP_00020745.2; ZP_00589771.1; YP_ 180376.1; NP_220072.1; 20 CAI27032.1; EAL87307.1; YP_112273.1; ZP_00376179.1; ZP_00498294.1; ZP_00492099.1; ZP_00463379.1; ZPJ30217095.1; CAI27980.1; AAK23707.1; CAD60736.1; ZP_00268854.1; E? A77706.1; ZP_00629856.1; NP_879905.1; EAA51976.1; NP_885384.1; CAI29613.1; AAA91879.1; ZP_00376555.1; ZP_00141283.2; NP_253516.1; NP_300890.1; AAB40885.1; AAN03814.1; ZP_00644737.1; AA036548.1; AAP98791.1; YP_079735.1; NP_388690.1; 10 AAP05672.1; NP_966507.1; P95596; EAN04065.1; NP 32124.1; ZP_00507305.1; NP_948204.1; ZP_00557093.1; YP_220287.1; ZP_00642506.1; ZP_00591535.1; NP 02193.1; NP_771418.1; AAA19188.1; AAK72471.1; 15 AA 72470.1; NP_345630.1; NP_756887.1; ZP_00404212.1; AAK72472.1; AAN50085.1; CAC46029.1; YP_001129.1; AAL64341.1; ZP_00526430.1; ZP_00308867.1; YP_053282.1; YP_036862.1; ZP_00210426.1; 20 ZP_00625423.1; ZP_00601791.1; YP_045732.1; YP_016277.1; P54533; AAV95488.1; AAW71149.1; YP_021029.1; YP_153983.1; ZP_00240355.1; XP 95801.2; EAN08156.1; ?? P94898.1; NP_326592.1; ?? P 1 1076.1; ZP_00239726.1; NP 980528.1; ZP_OO620223.1; ZP_00512893.1; YP H9413.1; NF 48088.1; XP_678378.1; YP_180009.1; NP_735347.1; CAI26632.1; AAN30046.1; BAB04498.1; AAN57909.1; ZP_00373647.1; NP_692788.1; 10 ZP_00053288.1; EAM72947.1; YP_078075.1; ZP_00006401.1; EAA16706.1; YP_221832.1; XP_742153.1; ?? L52038.1; NP_966125.1; YP_247286.1; AAA74473.1; ZP_00589476.1; CAI38117.1; A? 078292.1; EAA26057.1; BAD11090.1; ZP_00545191.1; CAE73952.1; YP_060098.1; BAB05544.1; NP_802451.1; NP 60876.1; AAL97648.1; ZP_00154188.2; ZP_00331725.1; AAV62625.1; CAG35032.1; YP 084091.1; ZP_00366080.1; 20 YP_139515.1; YP_121481.1; ZP_00340821.1; ZP_00531539.1; ZP_00630106.1; CAB06298.1; 0557324.1; CAE11227.1; AAX88061.1; ZP _00155680.2; NP_439206.1; ZP_00557867. 1;? AG56574.1; ZP_00156893.2; NP_7538 72.1;? AB06233.1; ZP_00557301.1; NP_80 5734.1; ZP_00053862.2; AAL21424.1; NP_ 752960.1; AAX64824.1; AAL19899.1; NP_ 309006.2; CAB50383.1; P18775; EAM9467 7.1; ZP_00550524.1; AAX66433.1; AAL 19 562.1; ZP_00600783.1; BAB34402.1; AAC 73980.1; YP_074347.1; AAG55381.1; NP_8 36552.1; NP_753873.1; ZP_00509629.1; E AM95020.1; AAG56575.1; ZP_00557953.1; BAB35717.1;? AX64548.1; C? E09880.1; AAX65422.1;? AC74660.1; P77783; YP_1 50613.1; ZP_00550244.1; AAL20417.1; NP _805214.1; YP_075885.1; NP_805215.1;? AX65421.1; NP_106243.1; YP_151327.1; A AX68089.1; BAB36807.1; ZP_00559319.1; AAG57632.1; AAL23129.1; CAB49710.1; NP_463170.2; CAG74160.1; BAB65038.1; YP_160885.1; YP_079335.1; NP_805998.1; YP_119157.1;; sulfite converts AAL21442 YP_149649.1; NP_804183.1; AAX66448.1; sulfite reductase to AAK79480.1; BAB81146.1; AA036949.1; anaerobic sulfide ZP_00576801.1; BAB81244.1; ZP_001451 subunit A 79.1; ZP_00662254.1; ZP_00575186.1; ZP_ R (74) 00536228.1; B? D86261.1; AAL81453.1; N Dsr? PJ 43176.1; ZP_00667024.1; NP_951149.1 49781.1; CAB49861.1; ZP_00511991.1; NP _143177.1; ZP_00416521.1; ZP_00528292. 1; AAL81454.1; NP_662769.1; YP_124839. 1; YP_127719.1; ZP_00532276.1; YP_0964 76.1; ZP_0051 1237.1; ZP_00528313.1; ZP_00661878.1; AAU82615.1; ZP_00588475.1; NP_662138.1; ZP_00335592.1; ZP_00547 738.1; ZP_00417715.1; CAB50658.1; BAD 85995.1; AAL80312.1; ZP_00562249.1; O5 7738; ZP_00541622.1; NP_143791.1; AAM 04028.1; YP_022952.1; AAA23200.2; NP_6 33770.1; NP_988039.1; NP_971592.1; NP_ 11 1690.1; C? C11546.1; NP_069364.1; NP_248450.1; NP_632687.1; AAT38120.1; AA Q66178.1; AAM18706.1; AAC65704.1; B ? B80961.1; NP_971022.1; NP_613849.1; NP _622237.1; AAK80598.1; NP_622352.1; A? B85701.1; YP_147006.1; CAA51740.1; A AM07137.1; AAN58909.1; NP_229439.1; E AM94068.1; NP_267503.1; CAG37745.1; N P_623138.1; AAO36853.1; ZP_00541900.1; A? L94626.1; ZP_O0563837.1; ZP_O05395; 94.1; AA036887.1; CAB49859.1; Q8XL63; AAP10806.1; NP_389436.1; YP_020666.1; YP_078946.1; ZP_00143318.1;? AK99669 .1; Q8DQ38; NP_816202.1; NP_345444.1; N P_692413.1; ZP_00240193.1; YP_011687.1; ZP_00504646.1; ZP_00575869.1; AA0355 21.1; YP_055711.1; NP_758176.1; P56968; 1EP2; ZP_00520109.1; NP_815421.1; YP_1 81916.1; ZP_0O575375.1; YP_180792.1; YP _175828.1; ZP_00382098.l; ZP_00130793. 1; ZP_00401561.1; ZP_00561001.1; ZP_006 55811.1; NP_012221.1; AAO75998.1; NP_9 52806.1; AAQ97765.1; AAL81452.1; AAS5 1833.1; ZP_00128609.1; NP_531891.1; CA E71880.1; NP_946031.1; CAH00358.1; CA A37672.1; CAG35490.1; AAN68771.1; AA V52085.1; AAV62537.1; ZP_00389517.1; Z P_00535043.1; XP_550297.1; ZP_0066477 7.1; NP_465359.1; C? D 14793.1; XP_33054 8.1; AAN15927.1; ZP_00234145.1; P23312; AAA 18377.1; ZP_00591027.1; CAI47849. 1; AAA67175.1; BAA 13047.1; AA? 72422. 1; ZP_00503086.1; NP_057313.2; ZP_0040 1594.1; P39866; XP_396639.1; XP_623086. l; BAB55OO2.1; P39870; AAB66010.1; NP_ 471282.1; EAA51298.1;; EAN09251.1; CA A56696.1; AAC69483.1; AAC49460.1;? A F63450.1; ZP_00505244.1; C ?? 32217.1; A AU84695.1; AAF04811.1; AAB93308.1; N P_796190.1; CAH08182.1; AAG30576.1; AB39554.1 sulfite converts NP 804181 YP_149647.1; AAA99277.1; BAB81242.1; sulfite reductase a? A036947.1; B? B81 144.1; A? K79482.1; anaerobic sulfide ZP_00576803.1; CAA60228.1; NP_952404 subunit C .1 ¡AAM06806.1 £ P_00542975.1; ZP_0050 (R74) 3774.1; ZP_00561650.1; ZP_00560787.1; Z DsrC P_00535294.1; NP_635288.1; ZP_0014513 0.1; ZP_00576896.1; ZP_00563866.1; NP_2 47865.1; AAU83232.1; AA035782.1; NP_9 87198.1; NP_614083.1; AAB84786.1; AAM 06538.1; ZP_00540799.1; NP_632386.1; A AM04125.1; ZP_00558670.1; NP_614085. 1; ZP_00130988.2; AAM06540.1; NP_6323 84.1; NP_633866.1; ZP_00631188.1; ZP_00 541754.1; EAN05933.1; ZP_00056315.1; Z P_00562004.1; ZP_00667805.1; YP_14772 1.1; AAK49018.1; AAC 17127.1; NP_53439 4.1; AAK89517.1; BAB92078.1; NP_10410 1.1; AAQ18184.1; AAM73544.1; P17847; A AA60450.1; NP_247530.1; ZP_00623963.1; AAK22600.1; ZP_00303419.1; ZP_005360 25.1; AAP46170 .1; NP_442378.1; NP_9543 06.1; CAC49509.1; AAP79144.1; BAD1536 4.1; ZP_00576654.1; ZP_00575700.1; CAG 36393.1; ZP_00579652.1; NP_918873.1;; Z P_00556586.1; ZP_00333573.1; CA? 4694 0.1; B AD53072.1; CAA46942.1; BAD1536 3.1; CAA34893.1; ZP_00558945.1; NP_952 142.1; BAD15365.1; ZP_00535147.1; NP_9 24503.1; BAE06055.1; BAB55003.1; ZP_00 415177.1; YP_036299.1; AAM06247.1; ZP _00392410.1; ZPJ) 0674499.1; EAM75787. l; AAO38372.1; YP_018789.1; AAN31831. 1;? AN31830.1; ?? N13223.1; B? D93723. 1; YP_083541.1; ZP_00575701.1; BAA065 30.1; ZP_00571935.1; ZP_00534536.1; AA P09103.1; YP_010301.1; ZP_00500892.1; Z P_00496099.1; ZP_00486991.1; ZP_00482 951.1; ZP_00467711.1; ZP_00452175.1; YP _104614.1; ZP_00645283.1; CAH34502.1; ZP_00237633.1; ZP_00516217.1; BAB043 32.1; CAA42690.1; NP_632080.1; CAG355 27.1; AAU83223.1; ZP_00544005.1; EA03 5589.1; E? L90616.1; AAB09032.1; ZP_005 75732.1; ZP_00558782.1; NP_250472.1; NP _637372.1; ZP_00139438.2; ZP_00107422. 2; ZP_00294167.1; ZP_00265695.1; CAG36 396.1; EAA65575.1; ZP_00149626.2; ZP_0 0549111.1; ZP_00413634.1; ZP_00525511. 1; YP_15767U; YP_07772U; P_229093 .1; CAF 19045.1; YP_120776.1; ZP_002421 20.1; ZP_00507972.1; ZP_00667812.1; NP_ 388212.1;; ZP_00653668.1; YP_257799.1; CAE22413.1; P22944; ZP_00537258.1; YP_ 041840.1; YP_187201.1; NP_372924.1; CA G44104.1; B AC79016.1; NP_771211.1; YP _175117.1; CAG75891.1; CAD29755.1; ZP _00379608.1; AAL64294.1; ?? C 17122.1; ZP_00162550.1; EA035459.1; AAD20825. 1; CAC06Ó95.1; AAC46074.1; ZP_0056360 5.1; ZP_00268907.1; NP_522783.1; NP_882 747.1; NP_886946.1; ZP_00281075.1; NP_7 65533.1; YP_189546.1; C? F32236.1; ZP_0 0169751.1; ZP_00535570 .1; YP_171020.1; ZP_00563990.1; ZP_00402377.1; NP_2528 19.1; AAV68379.1; NP_883424.1; YP_2525 66.1; NP_887867.1; AAC46135.1; ZP_0020 5156.1; ZP_00541555.1; ZP_00592076.1; Y P_009627.1; ZP_00493035.1; ZP_0046731 ll; ZP_0O450531.1; ZP_OO411899.1; NP_6 13536.1; YP_111251.1; NP_28533J1; YP_ 011484.1; YP_105749.1; NP_987944.1; B84911.1; CAC09931.1; NP_768957.1; Q8 TYP4; CAA79655.1; ?? B28156.1; ZP_005 29166.1; NP_069756.1; YP_010816.1; AA M1813J1; NP_961142.1; AAA23383.1; ZP _00567851. 1; ZP_00397695.1; AAP08405. 1; NP_216907.1; YP_113108.1; NP_613552 .1; NP_070472.1; NP_635324.1; CAE08992 .1; AAO36006.1; AAK46756.1; 1 ZJ9; AAB5 0233.1; ZP_? 532427.1; ZP_0052l920.1;? AO07346.1; NP_937004.1; NP_952492.1; A AO61105.1; YP_011615.1; NP_881958.1; C AB69775.1; ZP_00215253.1; ZP_00130392 .2; ZP_00130682.2; ZP_00131127.1; ZP_00 592685.1; ZP_00558070.1; ZP_00546885.1; ZP_00465281.1; ZP_00454984.1; ZP_0066 8317.1; ZP_00665867.1; NP_614767.1; C? ? 43512.1; AAU83053.1; NP_248188.1; NP _682139.1; CAA40717.1; ZP_00510136.1; YP_018067.1;; ZP_00563478.1; ZP_00661 349.1; YP_045466.1; AAV68654.1; YP_035 640.1; NP_971884.1; CAC33947.1; ZP_000 48074.2; ZP_00050155.1; AAM03878.1; A AK81453.1; AAC78310.1; CAA86992.1; Z P_00497750.1; EAO36822. 1; AAO38151.1; NP_709140.2; NP_633643.1; NP_953151. 1; C ?? 32416.1; AAT47760.1; AAU83339. 1; AAT99257.1; BAC73373.1; NP_247490. 1; AAC76390.1; A? K78079.1; ?? G58473. 1; BAB37639.1; ZP_00541003.1; ZP_00503 951.1; ZP_00417521.1; ZP_00265988.1; ZP _00621912.1; P00202; NP_614213.1; NP_0 68995.1; NP_800565.1; AAB85622.1; AAM 07522.1; AAF11421.1; AAB02352.1; YP_0 82906.1; ZP_00166363.1; ZP_00560684.1; ZP_00510774.1; EAM24821.1; ZP_005518 13.1; ZP_00667348.1; NP_621874.1; YP_26 6101.1; NP_794611.1; C ?? 76373.1; CA? 7 6342.1; CAA08858.1; AAK78015.1; AAC4 7160.1; BAB80366.1; ZP_00562782.1; ZP_ 00544290.1; AAZ18451.1;; ZP_00511163.1; ZP_00462654.1; ZP_00653977.1; NP_214 766.1; NP_716642.1; YP_259653.1; YP_01 2009.1; YP_117628.1; CAG37108.1; AAU9 5491.1; CAC39231.1; CAB95043.1; AAM9 2180.1; ZP_00237342.1; AAK23103.1; NP_ 739254.1; CAC19472.1; NP_977868.1; AA S07951.1; ZP_00217308.1; ZP_00565232.1; ZP_00541377.1; NP_949048.1; ZP_00267 392.1; ZP_00419332.1; ZP_00513264.1; ZP _00486799.1; EA038190.1; ZP_00397962. 1; YP_276576.1; AAV94843.1; ?? V94139. 1; NP_622539.1; NP_799995.1; NP_440189 .1; CAA74092.1 £ P_00054379.1; AAK812 91.1; BAB96809.1; AAQ21342.1; AAY209 96.1; ZP_00130766.1; ZP_00108506.2; ZP_ 00541311.1; ZP_00542829.1; EAM28721.1; ZP_00658370.1; AAP99874.1; YP_132435 .1; NP_808180.1; NP_777779.1; NP_22909 6.1; AA061114.1; AAL23380.1; CAG3655 3.1; AAU95489.1; CAD16132.1; ZP_00053 119.1; AAM21749.1; ZP_00128864.2; ZP_0 0593542.1; ZP_00574875.1; ZP_00575487. 1; ZP_00543494.1; CAA48368.1; ZP_00595 533.1; ZP_00557356.1; ZP_00435740.1; A AO08247.1; EAA71154.1; EA035986.1; Y P_153414.1; YP_14985J1; NP_708163.1; NP_936256.1; NP_614526.1; YP_071090.1; YP_046560.1; AA061117.1;? A061110.1; CAG75920.1; CAA11230.1; CAA46941.1; AAV47410.1; CAA92206.1; C47159.1; BAA16109.1; ZP_00356596.1; ZP_003459 38.1; ZP_00576139.1; ZP_00426552.1; ZP_ 00263533.1; EAN06682.1; AAY38236.1; E? M23497.1; ZP_00504882.1; ZP_0049839 0.1; ZP_00493060.1; ZP_00469245.1; ZP_0 0450851.1; ZP_00668314.1; YP_275286.1; YP_146312.1; NP_613608.1; AAZ15776.1; NP_251334.1; NP_632784.1; YP_102259.1; NP_793155.1; XP_759995.1; YP_261004. 1; CAH09344.1; CAC41649.1; ZP_0005653 1.1; AA075724.1; AAM22202.1 AAL8957 1.1; AAN69709.1; AAT50267.1; AAU1423 5.1; YP_101168.1; ZP_00576374.1; ZP_005 64969.1; ZP_00540769.1; ZP_00504835.1; P38681; Q01700; ZP_00397512.1; ZP_0039 7295.1; YP_174119.1; AAO38143.1; NP_70 7572.1; NP_623466.1; YP_221069.1; NP_0 69003.1; NP_251596.1; NP_800497.1; NP_ 988812.1; NP_753963.1; AAN29231.1; CA D71547.1; B AD84891.1; AAL52820.1; AA B85352.1; NP_962636.1; XP_324077.1; A? G23566.1; AAF87215.1; BAB55574.1; ZP_ 00562084.1; ZP_00563739.1; ZP_0054099 0.1; ZP_00543828.1; ZP_00537084.1; ZP_0 0537423.1; CAA42917.1; Q59110; AAQ798 21.1; AAD54888.1; YP_074057.1; NP_0692 60.1; NP_249130.1; NP_634587.1; NP_954 288.1; NP_952717.1; AAO61104.1; AAV68 690.1; AAU95493.1; CAB95039.1 AAP05283.1;; AAM38916.1; ZP_00574687.1; NP_948974.1; YP 99106.1;; ZP_00131780.2 ZP_00123638.1; NP_773400. l; YP_129655.1;; YP_236060.1; AAK03633.1;; AAP95731.1; CAD72806.1;; NP_792913.1; YP_223238.1;; NP_541491.1; AAX87606.1;; NP_531301.1; AAN33959.1;; AA 86411.1; NP_438715.1;; ZP_00156377.2 ZP_00265291.1; ZP_00416738.1 C? C45276.1;;? AN69632.1; AAD12043.1; AAC65465.1; ZP_00128741.1; YP_260249.1; AA076328.1;; AAN70916.1; YP_014595.1;; NP_471419.1; CAG75379.1;; YP_115360.1; NP_840485.1;; CAH07618.1; NP_219689.1;; YP_070566.1; NP_465502.1;; NP_798089.1; AAQ57824.1;; NP_107009.1; ZP_00596515.1; ZP_00278393.1 NP_669553.1;; CA? 52858.1 CAC90878.1;; NP_754157.1; AAG56842.1;; YP_150270.1; NP_929382.1;; AAX65797.1; NP_934398.1;; NP_837434.1; YP_206426.1;; AAO11031.1; EAO 17699.1; ZP_00637092.1; NP_804814.1;; ?? W29927.1 ZP_00005413.2; YP_112566.1; AAW29929.1;? AV96269.1; YP 079709.1;; AAL 14620.1 ZP 00498362.1; ZP_00472038.1 AAW29926.1; ZP_00633694.1 YP_148187.1;; AAA24775.1 ZP_00587566.1; ZP_00446786.1 NP_718076.1;; NP_814740.1; ZP_00350648.1; NP_254126.1; ?? F96793.1;; NP_390266.1; ZP_00423718.1; ZP_00583219.1 ZP_00303587.1; EAN10723.1 ZP_00217377.1; ZP_00568967.1 ZP_00455268.1; YP_269001.1 YP 175420.1;; NP_786078.1; CAF25793.1;; NP_251873.1; ZP_00334099.1; ZP_00280824.1 ZP_00218486.1; ZP_00166002.1 ZP_00579866.1; ZP_00316750.1 AAV95319.1;; ZP_00264504.1 ZP_00555465.1; ZP_00462564.1 NP_660655.1;; ZP_00152367.1 BAA90547.1; ZP_00629685.1 ZP_00384027.1; ZP_00285042.1 NP_693860.1;; EAM33140.1 YP_033231.1;; NP_662750.1; AAN66647.1;; YP_234212.1; NP_878735.1;; YP_261694.1; ? AB91531.1;; NP_791129.1; CAC 14908.1 ¡AU07486.1; ZP_00509653.1; ZP_00136528.2 ZP_00620998.1; NP_268377.1; ZP W643038.1; ZP_00564303.1 CAJ07708.1; AAK24030.1; CAB84837.1;; EAO 18485.1 YP_188644.1;; ZP_00415411.1 AAF41756.1;; ZP_00416202.1 NP 764743.1;; NP_523118.1; YP_040979.1;; ZP_0O588223.1 ZP_00590972.1; YP_037489.1; AAM64291.1;; YP_084670.1; ZP_00235566.1; AAL76389.1 YP_029439.1;; ZP_00530891.1 YP_020068.2;; ZP_00377677.1 ZP_00419457.1; YP_125825.1; 15 YP_094460.1;; YP_122821.1; ZP_00528275.1; ZP_00660845.1 AAM64228.1; AAW89435.1; C? A59012.1; ZP_00051756.1 NP_979712.1;; P21907; NP_240142.1; CAB52708.1; AA037825.1 CAA54841.1; Q42919; EAL92729.1; YP_253326.1;; CAB52675.1 CAA04696.1; AA042879.1; ZP_00385429.1; BAC23041.1 ?? D11426.1; ?? 036382.1; YP_ 190594.1;; B ?? 97662.1 CAA03939.1; B? A97664.1; AAF87216.1 AAM64230.1; BAB96757.1 CAA67782.1; XP_468660.1; BAA97663.1; CAA61194.1 AAQ02671.1; XP_477654.1; CAA58825.1 CAA54840.1 CAA59011.1; CAA97412.1; CAC05439.1; NP_196815.2; 10 C? E62054.1; XP_466575.1; AAB69317.1; CAA52442.1 CAA04994.1; AAL57678.1; C? B52674.1 CAA04993.1; BAB02125.1; ZP_00323827.1; CAA58775.1 AAM98087.1; BAD08586.1 NP_173838.1;; AAK99925.1; EAN98209.1; NP_345708.1; 15 EAN77674.1; AAL57688.1; AAB25541.1 ZP_00511972.1; XP_644814.1; XP_472942.1;; CAC07816.1 CAA04992.1; AAB69319.1 CAB52681.1; NP 777918.1; AAW44738.1;; EAL04742.1 20 EAL04547.1; EAA46705.1; ZP_00659395.1; EAA70588.1; ZP_00332697.1; CAB52685.1 AAS50565.1; AAB69318.1; AAZ23850.1 ?? L79959.1; ?? W24823.1 C ?? 49834.1; C? G86200.1; XP 761077.1; AAA34619.1 ZP 00110439.1; CAG07451.1 NP 535313.1;; NP_014158.1; AAT93017.1; O55044; NP_001017312.1; CAG79872.1;; NP_032088.1;; Q00612; AAH59324.1;; BAD17912.1 XP_331503.1;; ZP_00110078.1 NP_058702.1;; XP_311452.2;; Q29492 NP_105132.1;; ZP_00063705.1 BAD17947.1; CAG60989.1;; P11413; AAP36661.1; AAL27011.1;; 2BHL; ZP_00161394.2; NP_000393.2;; 1QKI; AAA92653.1;; AAA52500.1; XP_538209.1;; ZP_00644450.1 AAA41179.1; AAA63175.1; 15 XPJ07095.2;; CAA03941.1 XP_699168.1;; AAN76409.1; ZP_00319846.1; ?? N76408.1; AAW82643.1; ?? N76413.1; AAB29395.1; BAD17951.1; Q27638 EAL31619.1; CAB57419.1; BAD94743.1 P11411; NP_062341.1;; BAD17934.1 1H9B;; 1H94;; 1DPG;; AAB96363.1 BAD17891.1; BAD17877.1 BAD17941.1; BAD17905.1; 1E7Y; CAA58590.2; AAF48999.1; A? B02812.1; A? B02811.1;? F49000.2; BAD 17954.1; 2DPG;; NP_961605.1; AAB02809.1; AAA99073.1 AAK45410.1;; AAK93503.1 BAD17927.1; BAD17920.1 YP_177789.1;; BAD17898.1 NP_637497.1;; YP_193335.1; BAD17884.1; YP_200953.1; AAM36928.1;; AAF19030.2 ZP_00464292.1; CAB08746.1; 10 A? M51346.1;; XP_579385.1 AAA51463.1; AAM64231.1 ZP_00404158.1; NP_778577.1; AAC33202.1; NP_298355.1; ZP_0O651890.1; C? D28863.1 AAM64229.1; CAD28862.1 CAD43148.1; ZP_00046060.1 NP_964496.1;; ZP_00387215.1 XP_583628.1;; XP_559252.1; ??? 52499.1; C? D97761.1 AAR12945.1; AAR12953.1? AR12952.1; AAR12943.1 AAR 12946.1; CAGO4059.1 ZP_00413080.1; CAA19129.1; CAA03940.1; NP_960621.1; 20 EAM75831.1; CAB16743.1; CAES 1228.1; CAE51222X; AAG28730.1 AAG28728.1; AAA57029.1 A ?? 57025.1; CAE51229.1 ZP_00380754.1; E? M75226.1 AA019918.1; AA019916.1 AA019914.1; AA019917.1 XP_769603.1 CAC24715.1 EAA18517. 1; NP_213347.1; AAR26303.1; XP_680237.1; AAA65930.1;; NP_702400.1; NP_223744.1;; AAD08144.1; EAN31303.1;; NP_649376.2; EAN81514.1; ZP_00048966.1 10 AAG23802.1; CAI75777.1; AAV37033.1 NP 737152.1;; AAF24764.1 XP_233688.3;; AAS87299.1 NP_775547.2;; ??? H42677.1; CAH18137.1;; XP_425746.1; CAC27532.1; BAA82155.1 AAH81559.1;; NP_004276.2; 15 CAA10071.1;; ZP_00131371.2 ?? N06152.1; P56201; XP_697820.1; AAN06169.1; AAU95204.1 AAC08804.1 AAC08813.1 CAG06984.1; AAC08802.1 AAD35084.1;? AW81980.1; ZP_00572395.1; CAA45220.1 20 AAP44069.1; AAP44068.1; AAP44065.1 ; CAD77853.1; CAA90427.1; YP_037488.1; YP_181386.1; ZP_00235565.1; YP_084669.1; XP_476303.1; ZP_00282112.1; ??? N65341.1; 1ITZ; ZP_00231883.1; AAK78920.1; YP_020067.1; CAA86609.1; CAA86608.1; NP_662747.1; NP_925243.1; NP_734737. 1; YP_188491.1; AA035896.1; NP_764580. 1; NP_937158.1; AAM91794.1; AAK79316 .1; XP_471447.1; ZP_00328100.1; AAO29950.1; A? O07501.1; NP_687313.1 ; A? M62766.1; NP_935655.1; NP_682660.1; AAD10219.1; NP_670609.1; CAE06656.1; NP_801666.1; AAK34434.1; YP_203823.1; NP_566041.2; AAL98225.1; YP_071699. 1; NP_954463.1; NP_800691.1; YP_075950.1; YP_060741 1; CAE22131.1; NP_214208.1; AAF11802. 1; NP_893727.1; ZP_00527389.1; AAO09963.1; CAB82679.1;? AF93646.1; ?? F96525.1; YP_015228.1; ZP_00233073.1; ZP_00155697.1 NP 928282.1; ZP_00416485.1 NP 466182.1; ZP_00156879.2 ZP_00528273.1 20 NP_472138.1; NP_798983.1; NP_267781. 1; YP_131731.1; NP_439183.1; YP_131250 1; AAX88048.1; ZP_00585102.1 ZP_00381829.1 YP_206644.1; C? B58135.1 XP_550612.1; ZP_00129328.1 ZP_00530889.1; ZP_00132914.1 ZP_00537410.1? AQ00814.1; ZP_00123444.1 AA017218.1 CAG73773.1; AAT48155.1; BAB37233.1; NP_708699.2; NP_786741.1; 1QGD; NP_75 5395.1; CAG76812.1; ZP_00473037.1; AAK03722.1; AAG58065.1; ZP_00134256 .2;? AV61915.1; ZP_00389210.1; NP_949977.1; NP_790234.1; NP_249239. 1; XP_651488.1; XP_650850.1; YP_174605 .1; AAU36664.1; AAK03326.1; XP_650836 .1 AAQ57870.1; YP_237857.1; C? C18218. 1 YP_152097.1; CAC47341.1; C AA48166.1 AAX66924.1; ZP_00347807.1 15 ZP_00473067.1? AL21951.1; NP_346455.1; NP_359433.1 ZP_00418725.1 AAN58055.1; EAK85797.1 ZP_00398675.1 NP_840415 .1; ZP_00267930.1 20 AAB82634.2; XP_326821.1; BAB62078.1 YP_011742.1; NP_756618.1; ZP_0063165 5.1 YP_222392.1; AAL51492.1;? AN30626.1 E ?? 69343.1; ZP_00315920.1 EAA54486.1; ZP_00584101.1 ZP_00334879.1 AAP96482.1; NP_299218.1; YP_115427.1 YP_115433.1; YP_156595.1; YP_157602. 1; CAF26661.1; EAL90682.1; NP_534230.1; NP_779080.1; ZP_0004046; 3.1; YP_046678.1; NP_463872.1; ZP_0023007 3.1; YP_012971.1; C? D80256.1 10 ZP_00234258.1 NP_769223.1; ZP_00303056 .1 AAN70532.1; YP_199815.1; NP_469705.1? NP_784768.1; ZP_00038813.1 CAB82464.1 AAM38215.1; ZP_00145579.2 EAA65464.1; NP_716559.1; AAP86169.1; ZP_00283416.1 NP_638566.1; ZP_00264631.1; P21725 YP_262844.1; E? N07635.1 ZP_00281448.1; ZP_00566165.1 YP_034207.1; YP_125516.1; YP_094193. 1; ZP_00151666.2; ZP_00459681.1 ZP_00640383.1; ZP_00004561.2 20 ZP_00453223.1 CAG88854.1; NP_879793.1; NP_887926.1 AA091278.1; NP 83480.1; YP_122504. ] CAF32073.1 C? D16457.1; ZP_00635534.1 ZP_00629444.1? AL21368.1; 1AY0; AAB68125.1; ZP_00 376744.1; 1TKC; ZP_00243671.1; AAX66376.1; P29277; YP_ 149718.1; ZP_0 0270019.1; ? p_? 74448.1; NP_804254.1; NP_971914. 1; NP_708304.2; AAC75518.1; Q52723 CAH36963.1; ZP_00216610.1 ZP_00168684.2 10; NP_786435.1; NP_754872.1; CAA81260.1 CAA21881.1; AAK25582.1; BAB36750.1; ZP_00494674.1 YP_104015.1; ZP_00488354.1 CAG79209.1; ZP_00451919.1 AAF41816.1; NP_104787.1; CAH02329.1; EAL21160.1; ZP_00500476.1 15 ZP_00424086.1; ZP_00598759.1 ZP_00579602.1; AAA96746.2 ZP_00599356.1 AAG57574.1; CAA85074.1; ZP_00502496 1; ZP_00464206.1 AAW89704.1; NP_220269.1; AAS51554.1 CAB84897.1; CAA21989.1; EAK98686.1 CAF24238.1; AAW79357.1 20 ZP_00243955.1; AAX69269.1 CAB73633.1; ZP_00367926.1 XP_651838.1; CAH07360.1;? A075454.1 E? L17469.1; E? L20938.1 A? Q66751.1; E ?? 66976.1; ZP_00368598 1 EAA53893.1; XP_648840. 1; ZP_00120373 1; AAA26967.1; ZP_00050045.2 EAA64069.1; EAL17468.1 CAD25372.1; C? G88707.1; YP_115941.1 EAA70882.1; EAL86575.1; AAC83349.1 AAG59818.1; ZP_00514509.1 ZP_00645002.1 CAG84976.1; NP_326342.1; YP_016250.1 ZP_0002O488.2; NP_757837.1; EAA37632.1; EAN84471.1 NP_078425.1; NP_757463.1; ZP_0005375 6.1; CAA26276.1; NP_072728.1; AAB95721.1; AAP56380.1; BAA13834.1; AAN18173.1 ZP_00404457.1; CAH25336.1 ZP_00403049.1; ZP_00403048.1; ZP_00642219.1; ZP_00642210.1; ZP_00372877.1; ZP_00332380.1; A? M94004.1; BAA95691.1 AAG43112.1; ZP_00405278.1 20 ZP_0O053393.1; ZP_00374249.1 ZP_00120372.1; AAA50394.1 AAS93346.1; ZP_00642816.1 C? C21145.1; CAC21168.1; C? C21141.1 CAC2U61.1; C? C21159.1; CAC21148.1 CAC21142.1; CAC21140.1; AAS93351.1 AAK63242.1; AAK63239.1; AAS93349.1; AAS93347.1; CAC21156.1 AAS93356.1; CAC21157.1; CAC21162.1 CAC21137.1; CAE75672.1; CAA13584.1 CAA13374.1; AAK63244. 1 AAQ20076.1 ¡AAU95206.1 CAB60654.1; CAA13585.1 10; ZP_00405277.1; ZP_00510627.1 ZP_00374310.1; AAK63246.1 ZP_00574965.1 AAL94500.1; CAC11757.1; YP_076140.1; ZP_00534343.1 NP_228762.1; EAN81253.1: NP_953961.1; AAK17116.1 XP_651839.1; AAN50417.1; YP_077731.1 YP_090144.1; ZP_00207814.1 NP_247665.1; ZP_00559948.1; NP_346546.1; XP_734906.1 BAB80002.1; NP_111187.1; ZP_00403604 1; NP_687234.1; CAB71601.1 YP_023469.1; NP_532581.1; NP_988235. 1; AAX66248.1; C AB71607.1 20; CAB71595.1; CAB71613.1; CAB39235.1 Sulfate at YP_148197.1; ZP_00283191.1; NP_470749 sulfur .1; YP_079721.1; NP_464901.1; CAG74354 or .1; NP_928851.1; YP_056324.1; NP_39026 methylene- 7.2; YP_070081.1; NP_669932.1; ZP_00212 tetrahydro- 780.1; ZP_00424035.1; ZP_00462313.1; YP folate a _016771.1; YP_111755.1; AAO44589.1; NP methyl THF _992794.1; YP_034514.1; ZP_00390566.1; ZP_00236407.1; ZP_00232091.1; YP_0817 73.1; ZP_00494517.1; EAN09346.1; ZP_00 502057.1; ZP_00403921.1; NP_344902.1; Z P_00111860.1; NP_814782.1; AAL67561.1; AAA24492.1; AAG35235.1;; AAG35224.1; ?? G57088.1; B? B76974.l; ?? G35219.1; AAA24490.1; ZP_00163835.2; ZP_001581 00.1; AAA24208.1; AAO37703.1; YP_1754 22.1; AAV74553.1; AAV27335.1; AAA244 94.1; AAA23918.1; AAC75090.1; AAG352 18.1;; AAA24207.1; YP_172170.1; AAA24 495.1; AAA24493.1; AAG35221.1; ??? 24 209.1; NP_707923.1; AAG35223.1;? A? 24 489.1; P41576; AAD50492.1;? AV34527.1; A? L20985.1; AA? 24206.1; B ?? 28321.1; AAV74381.1; AAX65997.1; NP_804634.1; A? A24488.1; YP_150095.1; P37754; AAD 46733.1; BAA77736.1; NP_372035.1; P215 77; YP_040985.1; ZP_00384155.1; ZP_003 79330.1; YP_253321.1; NP_924063.1; NP_ 764747.1; NP_266778.1; ZP_00323177.1; N P_681366.1; P96789; ZP_00315559.1; AAC 43775.1; P41582; AAC43777.1;? AC43778 .1; ?? C43807.1; ?? C43817.1; P41580; ?? C43818.1; C43805.1; C43793.1; ZP_ 00326299.1; P41579; AAC43782.1; AAC43 811.1; AAC43808.1; AAC43800.1; AAC43 798.1; AAC43795.1; NP_785144.1; AAC43 806.1; AAC43804.1; AAC43781.1; CAD72 844.1; AAC43813.1; AAC43809.1; AAC43 803.1; AAC43787.1; P41578; P41574; AAC 43786.1; P41581; AAC43797.1; AAC43788.1; AAC43834.1; AAC43810.1;? AC43785. 1; ?? C43784.1; ?? C43812.1; P41577; ?? S99175.1; P41575; AAC43799.1; A? C4391 3.1; AAC43906.1; AAC43828.1; P41583; A AC43794.1; AAC43825.1; AAL76323.1; A AC43902.1; AAC43832.1; AAC43923.1; A AC43916.1; ?? C43914.1; ?? C43912.1;? AC43911.1; AAC43907.1; AAC43905.1;? AC43824.1; CAA41555.1; A? C43918.1;? AL27335.1; AAC43920.1; AAC43901.1; A? C43831.1;? C43904.1;? W29822.1; A AC43908.1; AAC43829.1; AAC43919.1; A AC43910.1; XP_342980.2; YP_1 14383.1; A AC43830.1; ZP_00319235.1; Q9DCD0; AA H59958.1; AAA74174.1; AAH11329.1; AA A74166.1; AAC43921.1; AAC43915.1; YP_ 081362.1; CAG32303.1; ZP_00564543.1; N P_694109.1; AAA74172.1; AAA74152.1; A AA74154.1; AAA74149.1; AAA74163.1; A ?? 74146.1;? AR24280.1; A ?? 74159.1; C? I95751.1; AAP88742.1; ?? L27345.1; ?? A74162.1; AAA74157.1; AAA74147.1; AA A27330.1; AAA74173.1; AAA74143.1; AA A74169.1; NP_391888.1; AAA74144.1; P52 207; AAA74167.1; AAA74164.1; AAA7415 8.1; NP_442035.1; A? L76320.1; AAA7530 2.1; AAA74171.1; ZP_00516323.1; AAL27 356.1; AAR97968.1; AAA74145.1; C? F230 41.1; CAA42751.1; AAA74151.1; 2PGD; ZP _00062611.2; A ?? 74156.1; YP_129657.1; ZP_00539485.1; AAK51690.1; AAA74161. 1; AAA74150.1;? AK64376.1; AAA74168. 1; AAA74175.1; AAA74165.1; XP_758724. 1; AAA74170.1; AAA74155.1; AAA74160. 1; CAG07546.1; AAA74153.1;? AQ13889. l; NP_910282.1; AAO32456.1; AAH44196. 1; CAA76734.1; AAF40494.1; AAP92648.1 ; AAQ13881.1; C? E46650.1; NP_998717.1; ??? O42814.1; ??? M64891.1; E? L03585.l; 013287; CAA94380.1; AAK49897.1; AAQ 13887.1; AAP33506.2; CAH02996.1; XP_6 25090.1; AAA74142.1; NP_798087.1; CAG 62903. 1; NP_012053.1; A? Ol 029.1; AAQ 13885.1; NP_934400.1; AAO 19944.1; AAO 19943.1; AAM78095.1; A? L76326.1;? AQ 13883.1; AAH95571.1; ZP_00524274.1; A AQ13879.1; CAG86870.1; CAG83189.1; A? C27703.1; NP_239940.1;; C? B61332.1;? ? O19942.1; A? L9941.l; ZP_00123635.1; CAB83570.1; AAO19934.1; AAQ13880.1; ZP_00131777.2; ZP_00135245.2; AAQ138 88.1; AAQ13882.1; EAL18227.1; NP_0117 72.1; BAD98151.1; NP_438711.1; AA0323 96.1; EAA48517.1; NP_777731.1; AA0324 97.1; AAU36620.1; AAM61057.1; AAX876 02.1; AAQ13878.1; CAE70848.1; BAD367 66.1; EAL88658.1; AAS53500.1; XP_33053 6.1; ZP_00156373.2; XP_313091.2; F96 795.1; CAE53864.1; AAB41553.1; CAA225 36.1; AAC27702.1; YP_206428.1; BAC063 28.1; EAA59263.1; EAL31500.1; AAP9572 8.1; CAB10974.1; AAQ13886.1; P70718; A AF45732.1;; AAK03638.1; AAC65319.1; E AA67653.1; AAR25841.1; ZP_00152366.1; AAB29396.1; EAN05374.1; P41573; YP_21 9832.1; CAD80254.1; CAD56883.1; C? C4 6511.1; AAP05178.1; NP_532215.1; ?? Q1 3884.1; NP_104453.1; AAL27320.1; NP_66 0459.1; AAO36383.1; AAO76329.1; CAH0 7617.1; YP_099135.1; NP_542102.1; YP_2 22920.1; AAF39196.1; AAB20377.1; ?? C9 7362.1; NP_878753.1; CAE07634.1; AAP9 8300.1; NP_892888.1; NP_228248.1; NP_6 ? P_15967TX;; NP_948972X; EAO 16453.1; AAW90238.1; CAB85348.1;; ?? Q58240.2;; Q9K139; ZP_00203717.1; ZP_00522015.1 NP_842140.1;; AAT08720.1 AAA17145.1; CAE11079.1; ZP_00648773.1; AAA17140.1 AAD08536.1;; CAD73047.1; NP_224106.1;; ZP_00369384.1 ZP_00367543.1; YP_178349.1; ?? L15878.1; ?? P77737.1; A? L15879.1; ?? L15872.1;? AL15875.1 ?? L15876.1; ?? L15874.1 ZP_00369996.1; AAL15880.1 ZP_00657668.1; YP_055164.1; ZP_00328326.1; ZP_00416486.1 CAD14933.1;; ZP_00280696.1 ZP_00107110.1; AAP05431.1;; NP_719093.1;; AAH09680.1; 15 AAH10103.1;; AAH18847.2; ZP_00587113.1; YP_172513.1; ZP_00263754.1; ZP_00165284.2 AA032594.1; AA 55523.1 YP_115432.1;; ZP_00 16112.1; P51778 NP_251486.1;; NP_791941.1; AAX66375.1;; NP_754871.1; 20 AA032544.1; AAT51194.1; AAS51032.1 NP_035658.1;; NP_804255.1; NP_440132.1;; NP_439282.1; ZP_00495461.1; NP_113999.2; YP_046627.1;; ?? L21367.1; ZP_00160100.2; ?? F96524.1; YP 49719.1;; XP_533146.1; NP_924543.1;; XP_420949.1; BAB74262.1;; XP_397306.2; ZP_00242010.1; ZP_00415676.1 AAP98016.1;; CAG73774.1; CAA18994.1;; ZP_00269391.1 AAF38500.1;; AAF39419.1; NP 92637.1;; ZP_00653780.1 NP_800690.1;; YP_220057.1; NP_219818.1;; AAX46381.1 A? G43169.1;; ZP_00133180.1 ZP_00498695.1; ZP_00451399.1 ZP_00635839.1; AAK03686.1;; ZP_00212331.1; CAA78965.1 ZP_00583402.1; ZP_00473621.1 CAG31705.1; AAZ19038.1; YP_234996.1;; NP_671009.1; ?? H61957.1;; C? C89319.1;; NP_681257.1;; AAH68191.1; AAH84118.1;; ZP_00156967.1 ZP_00509652.1; XP_306040.2;; NP_705968.2;; AAO07500.1; YP_149357.1;; NP_937157.1; AAX63913.1;; NP_751968.1; YP_202288.1;; Q8FLD1; 1I2R;; 1I2P; 10NR;; CAE08274.1;; ZP_00444183.1 ZP_00642538.1; C? G76785.1; E? K96114.1; 1I2N;; XP_760285.1 AAN67781.1;; ZP_00566285.1 ZP_00534289.1; ZP_00459221.1 YP_270759.1; YP_069148.1; NP_838015.1;; NP_228107.1; ZP_00640073.1; NP_534942.1; EAL18010.1; AAW46393.1; NP_001017131.1;; 1VPX; ZP_00167021.1; ZP_00135489.1 ZP_00561481.1; ZP_00314890.1 YP_017300.1;; ZP_00507474.1 EAA72420.1;; NP_463873.1; NP_883543.1;; NP_880193.1; NP_887991.1;; NP_636221.1; YP_206643.1;; XP_640977.1; AAX88128.1;; ZP_00551628.1 AAP95288.1;; YP_131732.1;; 1I2Q; ZP_00316642.1; AAP07679.1; ??? L60146.1; YP_259093.1; ?? U38962.1; AAM35790.1; AAK03723.1;; lI2O;; lUC; YP_170072.1;; NP_927918.1; NP 736284.1;; AA032444.1; Q9S0X4 CAF99897.1; AAW50031.1 ZP_00597354.1; NP_708303.1; ; NP_623494.1;; NP_247955.1; ZP_00422910.1; CAE21423.1; AAZ22459.1; NP_777717.1; C? G78269.1;; C? G35157.1; AAP99564.1;; AAN31490.1 CAA34078.1; NP_013458.1; NP_239926.1;; YP_144332.1; AAX15925.1; ZP_00473066.1 YP_149238.1;; NP_011557.1; AA032543.1; AAS56158.1 XP 20715.2;; NP_988428.1; CAG61370.1;; CAF24415.1; 10 ?? 75991.1; YP_081036.1; XP_508205.1;; XP_591134.1; CAG58006.1;; ZP_00346593.1 EAA66113.1;; BAB07504.1; BAC24729.1;; CAE58522.1 NP_954019.1;; NP_466265.1; AAX69845.1; YP_015318.1; 15 NP_660444.1;; AAL65638.1 ZP_00230153.1; AAW69342.1 ZP_00538489.1; ZP_00530466.1 AAP37846.1;; NP_878795.1; AAL65636.1; AAL65632.1; AAL65631.1 AAL65625.1; AAL65622.1;? AL65620.1 AAL65619.1; AAL65613.1; BAD94458.1 AAK79315.1;; AA034876.1;; Q899F3 YP 182117.1;; NP_213080.1; ÑP_660916.1;; XP_480152.1; AA032445.1; AAN87407.1 ZP_00576424.1;? AK93861.1; E? O20247.1; NP_786742.1; AAM66063.1; CAG89600.1; ZP_00053003.1; ZP_00357963.1 ZP_00523938.1; NP_801665.1; EAL91678.1;; YP_004676.1; BAB80398.1;; AAF10909.1;; Q9RUP6 CAA89874.1; E? 023438.1 NP_111185.1;; CAJ03645.1 NP_967077.1;; NP_391592.2; ??? G34725.1; ??? R10031.1 10 AAQ65460.1;; AAP10311.1; YP_177374.1;; NP_472214.1; AAM50780.1; AAF47106.2; ZP_00513183.1; AAQ17460.1 CAH08777.1;; E? N83889.1 YP_214732.1;; YP_100521.1; EAN81646.1; XP_329326.1; 15 ZP_00622649.1; ZP_00661024.1? A076765.1;; E 58088.1; EAA49912.1;; YP_020065.1; AAQ59919.1;; ZP_00402027.1 YP_037486.1;; NP_693926.1; YP_073895.1;; YP_010876.1; AAK15382.1; AAK15373.1 20 ZP_00302582.1; ZP_00554321.1 ZP_00397276.1; ZP_00332965.1 AAK15379.1; ZP_00577093.1 E? L86159.1;; CAC11755.1; ZP_00207691.1; ZP_00386434.1 YP_023467.1;; AAK25576.1; ZP_00266165.1; ZP_00311094.1 ZP_00592141.1; ZP_00588840.1 AAK15385.1; NP_816912.1; AAN49485.1;; AAK34644.1; AAN58723.1;; NP_802941.1; YP_253060.1;; AAM80284.1; ZP_00376041.1; YP_222463.1; ZP_00207851.1; XP_417183.1; EAA76445.1;; YP_041250.1; NP_372305.1;; YP_060987.1; AAL98495.1;; XP_417182.1; AAL80779.1;; ZP_00579645.1 EAM28893.1; YP_080940.1; YP_093370.1;; AAU21341.1 AAF39749.1;; AAL51426.1; NP 772111.1;; Q8WKN0; Q8SEL8 C? D27636.1; C? D27635.1 CAD27630.1; CAD27629.1 CAD27627.1; CAD27625.1 CAD27624.1; ZP_00215873.1 AAN02022.1;; ZP_00267913.1 AAS53358.1 transhydrogenase conversion ?? C76944 P27306; A? C43068.1; NP_756777.1; NP_7 udh (R70) redox of 09766.2; Q83MI1; Q8X727; AAG59164.1; nucleotides AAX67921.1; YP_153038.1; CAG77139.1; of pyrimidine YP_068670.1; NP_667661.1; NP_931901.1; NP_799321.1;? AF93328.1;? O09639.1; YP_205822.1; YP_131541.1; CAA46822.1; A? Z24633.1; YP_ 154714.1; ZP_00416263. 1; NP_251681.1; EA 24138.1; A? Y36945. 1; NP_791929.1; AAN67764.1; YP_274087. 1; YP_259077.1; O05139; ZP_00263769.1; CAD74394.1; ZP_00315937.1; YP_046885 .1; ZP_00653465.1; AAZ19183.1; YP_1697 00.1; AAW50006.1; NP_961763.1; ZP_005 74185.1; ZP_00546586.1; ZP_00523033.1; NP_217229.1; NP_532346.1; AAV97042.1; CAF23449.1; CAC46308.1; ZP_00622185. 1; NP_108478.1; ZP_00400087.1; P71317; Z P_00317524.1; ZP_00524635.1; YP_14355 3.1; YP_005669.1; EAN07674.1; ZP_00625 011.1; NP_105199.1; AAN30810.1;; AAL5 1327.1; EA037648.1; YP_222565.1; ZP_00 307577.1; ZP_00265019.1; AAN70931.1; C AF22875.1; C? C47627.1; CA? 39235.1; NP _213506. l; ZP_00284261.1; NP_945538.1; EAN27796.1; AAF34795.3; AAF79529.1; Z P_00141283.2; ZP_00492121.1; AAR2128 8.1; NP_253516.1; AAN03817.1; AAG1788 8.1; ZP_00449174.1; YP_180009.1; YP_034 342.1; NP_533297.1; CAI26632.1; NP_767 089. 1; AAC26053.1; YP_246823.1; ZP_004 97224.1; YP_067405.1; XP_331183.1; AAD 53185.1; E ?? 51976.1; NP_220840.1; ZP_0 0269527.1; ?? F34796.1; C ?? 11554.1; C? A44729.1; 1DXL; EAM25883.1; AAF95555.1; YP_115390.1; AAX88688.1; NP_439387 .1; YP_160845.1; EAK93183.1; ZP_001574 02.1; ZP_00464142.1; CAA70224.1; ZP_00 055963.2; EAO33154.1; ZP_00575798. 1; E AL87307.l; ZP_00211386.1; ZP_00340462 .1; AAR38073.1; NP_240038.1; AAS47493. 1; ZP_00317120.1; NP_298158.1; ZP_0021 0426.1; ZP_00637900.1; N23154.1; C? F26798.1; ZP_00154973.1; YP_170418.1; N P_779995.1; ZP_00151187.2; AAB30526.1; ZP_00526430.1; E ?? 77706.1; CAD60736.1; BAB05544.1; ZP_00153792.2; ?? U379 41.1; AAV93660.1; NP_969527.1; CAD149 73.1; ZP_00511405.1; NP_798896.1; EA03 0592.1; ZP_00665518.1; YP_265659.1; NP_ 925975.1; NP_388690.1; ZP_00633839.1;? ? V29309.1; C? G35032.1;? AK02977.1; C AF92514.1; ZP_00597315.1; C ?? 37631.1; ZP_00582828.1; AAK22329.1; 1 EBD; ZP_0 0545191.1; NP_662186.1; CAD72797.1; A AG12404.1; ZP_00601791.1; BAB44156.1; ZP_00644737.1; XP_475628.1; NP_360330 .1; AAA91879.1; AAC46405.1; ZP_001397 027l; YP_131302.1; CAF23812.1; ZP_0034 0821.1; EAA26462.1; ZP_00122566.1; ZP_00150164.2; ZP_00132373.2; YP_021029.1; NP_908725.1; C? G76686.1; A? O 10051.1; NP_935564. 1; NP_953492.1; CAB05249.2; ZP_00240355.1; ZP_00589771.1; ZP_0058 5786.1; NP_794013.1; XP_635122.1; YP_2 74206.1; NP_757897.1; NP_692788.1; NP_892685.1; AAP11076.1; CAG85768.1; NP_ 980528.1; NP_250715.1; YP_146914.1; CA Cl 4663.1; AAP96400.1; XP_758608.1; YP_ 001129.1; NP_767536.1; ZP_OO538550.1; P_069256.1; C ?? 71040.1; ZP_00597992.1; AAY37054.1; NP_706070.2; NP_752095.1; CAA24742.1; A? F49294.1; YP_205561.1; NP_999227.1 C? J08862.1; CAA71038.2; ZP_00283805.1; XP_320877.2; ZP_005287 40.1; YP_148949.1; YP_040992.1; NP_792 022.1; YP_002403.1; CAG81278.1; YP_085 309.1; AAS47708.1; ZP_00662383.1; EAN8 0618.1; ZP_00411894.1; YP_149503.1; YP_ 078853.1; NP_470744.1; YP_013986.1; XP _623438.1; ZP_00020745.2; ZP_00134358. 2; ZP_00536790.1; AAP10890.1; NP_80404 3.1; AAX64059.1; YP_186404.1; NP_37204 2.1; AAL19118.1; AAR38213.1; ZP_00507 305.1; EAO31379.1; ZP_00415841.1; CAA 49991.1; ZP_00589476.1; EAN90443.1; NP _716063.1; NP_777818.1; YP_134400.1; N P_464896.1; AAQ91233.1; ?? N50085.1; A? H44432.1; C? B06298.1; ZP_00233557.1; AAN69413.1; E? N96941.1; NP_930833.1; NP_266215.1; AAN48422.1; CAA72131.1; CAA61483.1; ZP_00383074.1; AAD55376. 1; YP_020826.1; NP_885384.1; NP_879905 .1; CAA62982.1; CAE46806.1; YP_247286. 1; AAR10425.1; AAV48381.1; CAA67822. 1; NP_635936.1; EAA26057.1; AAM38502. 1; CAE46804.1; NP_389344.1; AAH56016. 1; YP_253312.1; YP_257414.1; YP_260503 .1; C ?? 72132.1; C? G31211.1; YP_199361. 1; ZP_00166998.2; ZP_00565931.1; NP_84 2161.1; ZP_00591535.1; ZP_00499160.1; N P_221155.1; NP_660554.1; AAS53883.1; N P_815369.1; CAG58981.1; ZP_00154188.2; ZP_00554136.1;; AAP98791.1; AAM93255 .1; EAN04065.1; YP_180376.1; NP_953634 .1; NP_300890.1; CAI27032.1; AAR38090. 1; EA031664.1; ZP_00595215.1; ZP_00661 894.1; YP_078075.1; C? R27980.1;; ZP_003 96676.1; AAN75618.1; YP_253771.1; NP_2 20072.1; CAB84783.1; EAM31433.1; ZP_0 0263252.1; YP_019413 .1; A? N75159.1; C AA62435.1; NP_778978.1; AAV28779.1; N P_360876.1; NP_031887.2; NP_878457.1; NP_966125.1; AAC53170.1; ?? H18696.1; AAH62069.1; AAW71149.1; CAA59171.1; AAN69768.1; AAF39644.1; ZP_00373647. 1; EAL29693.1; NP_763640.1; A? A35764. 1; NP_116635.1; YP_148232.1; ?? V28746. 1; AAW89295.1; AAN05019.1; YP_084091 .1; 1V59; P31052; ZP_00245417.1; ZP_0021 2990.1; ZP_00266952.1; AAF41719.1; NP_ 298837.1; NP_623271.1; AAN75183.1; NP_ 883660.1; NP_888964.1; AAN15202.1; AA U45403.1; ZP_00308867.1; BAD92940.1; N P_000099.1; 1 ZMD; ZP_00418304.1; ZP_0 0399987.1; AAO90013.1; NP_764754.1; AND P _258846.1; B? E00452.1; ZP_O0305550.1; ZP_00210841.1; 3LAD; YP_188656.1; NP_ 880776.1; ZP_00007570.1; B AD11090.1; A AW89611.1; EAN09173.1; AAY38013.1; A AF41363.1; ZP_00670517.1; AAN75720.1; CAB84413.1; AAM36402.1; NP_966507.1; AAC44345.1; ZP_00107537.1 CAA61894. 1; CAA57206.1; CAB65609.1; ZP_0057846 3.1; ZP_00550077.1; YP_156710.1; CAD61 860.1; BAD11095.1; ZP_00537692.1; NP_7 64349.1; NP_792904.1; AAN00129.1; AAB 88282.1; 1BHY; 1 OJT; ZP_00245307.1; ?? A83977.1; ZP_00592008.1; ZP_00557093. 1; AAQ58205.1; ZP_00669696.1; AAN0088 2.1; C? A54878.1; YP_200681.1; ?? Q5874 9.1; YP_274470.1; YP_154852.1; NP_7639 20.1; YP_079735.1; NP_879789.1; NP_889 077.1; YP_067730.1; NP_636857.1;? 078 292.1; B? B33285.1; C? H93405.1; NP_842 316.1;? D30450.1; ZP_00170705. 2; ?? F 99445.1; AABO 1381.1; ZP_00245305.1; AA W71147.1; NP_250278.1; CAD15305.1; A AK23707.1; P_00160593.1; AA044599.1; NP_975267.1; CAH00655.1; ZP_0021274 7.1; CAE20510.1; YP_040483.1; ZP_00474 314.1; ZP_00464633. 1; NP_789199.1; AAP 05672.1; AAV47687.1; ZP_00108447.1; ZP _00516811.1; ZP_00467577.1; ZP_004511 58.1; NP_883762.1; YP_187838.1; ZP_0037 6179.1; BAB06371.1; BAC24467.1; NP_76 3632.1; YP_036862.1; AAA99234.1; 1LPF; ZP_00579524.1; ZP_00561492.1; ZP_0050 0723.1; ZP_0O486500.1; YP_220287.1; ZP_00239726.1; NP_681658.1; NP_893415.1; E AN08634.1; ZP_00578482.1; ZP_00531539 .1; ZP_00463093.1; ZP_00397330.1; ZP_00 642506.1; ZP_00620223.1; YP_126870.1; C? C46029.1; ?? P94898.1; CAE08145.1; ZP _ 00512893.1; YP_123783.1; ZP_00401182. 1; NP_148088.1; ZP_00207996.1; AAF6413 8.1; AAA96487.1; EAM93501.1; ZP_00463 487.1; YP_074243.1; YP_095531.1; NP_73 4581.1; NP_692336.1; NP_979105.1; YP_1 16095.1; ZP_00630106.1; AAP95326.1; CA A61895.1; AAL00246.1 transhydrogenase conversion NP 334574 NP_962508.1; NP_302686.1; YP_117224.1 pntB (R70) redox from; ZP_00517957.1; ZP_00112135.1; NP_681 nucleotides 485.1; ZP_00203498.1; ZP_00411543.1; BA pyrimidine B75107.1; ZP_00673742.1; ZP_00315150.1; AAZ25627.1; EAN07189.1; ZP_00549520 .1; NP_105891.1; C? C47439.1; ZP_003187 02.1; ZP_00400206.1; C? D77499.1; ZP_00 164663.2; YP_170777.1; YP_005747.1; YP 43474.1; ZP_00626042 .1; YP_190751.1; AAQ87239.1; NP_773764.1; Q59765; NP_9 49516.1; YP_223058.1; AAK25265.1; AAQ 57778.1; ZP_00348709.1; YP_266000.1; ZP _00267648.1; ?? N47534.1; ?? M35812.1; ZP_00241933.1; ZP_00523138.1; C? E213 39.1; ZP_00577769.1; YP_159578.1; ZP_00 417258.;; ZP_OO599375.1; ZP_00377317.1; ZP_00314523.1; NP_719280.1; NP_79518 8.1; NP_888489.1; NP_884728 .1; YP_2572 65.1; YP_202268.1; ZP_00215077.1; ZP_00 051959.2; ZP_00420704.1; ZP_00262288.1; NP_840935.1; CAB84437.1; ZP_00600539 .1; ZP_00507651.1; NP_248887.1; NP_6960 33.1; EAN27201. 1; ??? Y40045.1; AAN657 89.1; YP_126266.1; ZP_00303636.1; ??? N6 2246.1; NP_893262.1; YP_094909.1; YP_1 23265.1; C? D16440.1; YP_115164.1; C? E 07209T; W90? T2.TJ? AF4? 382.1; YP_2 77177.1; CAG75106.1; ZP_004978 1.1; NP _929428.1; NP_440860.1; ZP_00339822.1; YP_246075.1; ZP_00452010.1; ZP_002166 76.1; ZP_00350599.1; AAQ00284.1; AAO9 1446.1; ZP_00655137.1; AAZ18418.1; ZP_ 00464938.1; ZP_00457884.1; ZP_0066920 7.1; YP_047599.1; YP_070739.1; NP_8004 31.1; AAF96466.1; NP_359741.1; YP_0670 26.1; ZP_00598774.1; ZP_00168669.2; YP_ 206543.1; YP_132575.1; ZP_00170342.1; Y P_063030.1; ZP_00282750.1; EAA25826.1; NP_936867.1; ZP_00634798.1; ZP_00153 170.2; ZP_00585179.1; NP_220468.1; AAX 65404.1; NP_707499.1; YP_150630.1; AAZ 27182.1; ZP_00581426.1; ZP_00170332.1; ZP_00278457.1; ZP_00170182.2; ZP_0016 7836.2; NP_533165.1; AAK02836.1; ZP_00 135326.1; NP_439514.1; ZP_00157199.1; Z P_00638575.1; AAX88574.1; BAC68112.1; YP_055340.1; CAC 16724.1; ZP_00169560 .2; ?? U37830.1; ZP_00006258.1; ZP_0062 8636.1; AAQ87370.1; ZP_00133010.1; AA Q66400.1; ZP_00657269.1; ?? P96434.1; Z P_00166114. 2; ZP_00414377.1; E? M2433 1.1 AAV96060.1; CAE68875.1; ZP_00620 416.1; ZP_00554983.1; AAB52670.1; EAA 53262.1; EAL86026.1; ZP_00644761.1; ZP _00149697.1; XP_312859.2; CAB88572.2; XP_326633.1;; CAF99856.1; CAF99322.1; ? p_424784.1; NP_032736.2;? AH08518.1; BAC39226.1;? AH91271.1; C ?? 89065.1; ZP_00048453.1; AAF72982.2; XP_536481. 1; AAH66499.1; BAC39564.1; BAC30596. 1;; ZP_00533817.1; AAH81117.1; NP_7763 68.1; AAA21440.1; P11024; AAC43725.1; Ql 3423; CAD38536.1; XP_679831.1; CAH 90079.1; CAA90428.1; ZP_00054747.1; C41577.2; EAA18012.1; NP_702397.1; EA K89427.1; CAA 10358.1; AAC51914.1; AA K18179.1; NP_971796.1; 1NM5; NP_52251 5.1; 1PTJ; AAG02246.2; 1XLT; 1PNQ; EAK 88482.1; XP_666495.1; XP_646840.1; EAA 77364.1; XP_648285.1; XP_666155.1; AAH 32370.1; lD4O; XP_669801.1; XP_694555. 1; XP_517776.1; AAO07275.1; EAA58767. 1; AAO07276.1; AAP50917.1; AAP50916.1 ; AAP15452.1; AAP50915.1; AAP50914.1; XP_695634.1; ??? 29081.1; NP_522512.1; ZP_00202835.1; ZP_00208470.1;? D099 42.1; XP_738089.1; XP_653216.1; CAI374 45.1; XP_582741.1; ZP_00655886.1; ZP_00 166345.1; ZP_00048483.2; ZP_00675280.1; ZP_00412076.1; ZP_00170534.2; B? B052 58.1; XP_422112.1; AAB23106.1; ZP_0066 5383.1; ZP_00544728.1; ZP_00385815.1; Z P_00674068.1; ZP_00634699.1; NP_99062 2.1 Transhydrogenase conversion NP 214669 NP_962506.1; NP_302688.1; YP_117222.1 pntA (R70) redox from; ZP_00673740.1; ZP_00400208.1; YP_202 Nucleotides 272.1; YP_005749.1; ZP_00162920 .2; Pyrimidine ZP_0 0315152.1; BAB75109.1; NP_681483.1; ZP _00150890.1; ZP_00507653.1; ZP_005495 18.1; ZP_00203569.1; YP_159576.1; ZP_00 112137.1; YP_190749.1; ZP_00417256.1; C? D16437 .1; ZP_00523136.1; N62244.1; YP_103924.1; ZP_00282748.1; NP_24888 5.1; ZP_00140616.1; ZP_00669209.1; AAY 40043.1; ZP_00485169.1; A? 091444.1; ZP _00517955.1; ZP_00241930. 1; ZP_001703 40.1; CAC16725.1; YP_277175.1; NP_5225 10.1; YP_257267.1; NP_881481.1; E532 62.1; ZP_00166078.2; NP_884730.1; ZP_00 167837.2; NP_840933.1; ZP_00262286.1; Z P_00464936.1; ZP_00497829.1; YP_17077 9.1; ZP_00411541.1; ZP_00216674.1; CAE 21341.1; ZP_00492956.1; ZP_00170330.2; ZP_00168666.1; AAN47251.1 AAM35806 .1; BAC68113.1; CAE07207.1; ZP_0027845 9.1;? AQ57780.1; YP_1 15165.1; XP_32663 3.1; ZP_00598776.1; ZP_00170183.2; AAQ 87237.1; C? B88572.2; ZP_00166343.1; ?? Q00286.1; C? D77501.1; ZP_00314525.1; E? L86026.1; NP_105889.1; YP_055339.1; N P_773766.1; EAN07187.1; EAN27202.1; C AB 84439.1; ZP_00318700.1; ZP_00414378; .1; C? G75105.1;? AB52670.1; ZP_002676 50.1; 1 PTJ; 1 L7D; EA? 77364.1; XP_562937 .1; ZP_00600541.1; AAK25267.1; CAE688 75.1; ZP_00420705.1; NP_949518.1; AAW 90110.1; 1NM5; ZP_00377320.1; NP_8932 64.1; ZP_00169561, 2; NP_696034.1; NP_92 9427.1; YP_247378.1; AAF41384.1; NP_54 1301.1; YP_070740.1; NP_669446.1; CAC4 7441.1; AAN34144.1; ZP_00133009.1; ZP_ 00122250.1; ZP_00638574.1; YP_223056.1 ; ZP_00208471.1; ZP_00626044.1; A? K028 37.1; AAZ18416.1; CAA46884.1; NP_7538 90.1; NP_360971.1; ZP_00154275.2; AAP9 6435.1; AAX88575.1; XP_646840.1; ZP_00 655135.1; AAU37831.1; 1F8G; ZP_005851 78.1; YP_150631.1; NP_805194.1; AAL203 98.1; NP_439513.1; NP_707500.2; YP_047 601.1; ZP_00628637.1; EAM24330.1; AAF 96465.1; NP_800432.1; NP_440856.1; AAK 00588.1; NP_719279.1;? AG56590.1; NP_2 21211.1; ZP_00577994.1; ZP_00340908.1; ZP_00581427.1; AAZ27926.1; YP_063028 1; AAH66499.1; YP_206544.1; YP_132574 1; A? V96061.1; NP_533164.1; ZP_000062 59.2; AAH91271.1; AAQ87369.1; ZP_0021 5076.1; NP_032736.2; AAH08518.1; CAA8 9065.1; AAF72982.2; YP_067788.1; ZP_00 303638.1; AAK18179.1; ZP_00634797'.1; Z P_00135325.1; CAF99322.1; O07277.1; NP_936868.1; ZP_00657268.1; ZP_005549 82.1; AAG02246.2; XP_536481.1; NP_7763 68.1; C? D38536.1; P11024; AAA21440.1; Q 13423; XP_424784.1; AAH81117.1; AAC 51914.1; CAA90428.1; YP_126268.1; ZP_0 0620417.1; YP_094911.1; CAF99856.1; CA H90079.1; YP_265998.1; XP_666495.1; EA K89427.1;; AAC41577.2; ZP_00120256.2; AAA29081.1; AAA61928.1; ZP_00659346. 1; ZP_00599372.1; XP_666155.1; EAK8848 2.1; B AC39226.1; BAC30596.1; BAC39564 .1; AAB81400.1; ZP_00599373.1; NP_7023 97.1; EAA18012.1; XP_603436.1; XP_6798 31.1; XP_517777.1; AAM44187.1; NP_217 296.1; AAK47169.1; YP_080482.1;; CAA4 4791.1; NP_302068.1; CAE07016 .1; NP_69 4147.1; ZP_00539848.1; ZP_00386339.1; A AM44190.1; XP_598602.1; ZP_00293711. 1; XP_740533.1; E? M73707.1; YP_117835, 1; YP_126314.1; ZP_00526155.1; BAB060 48.1; YP_094958.1; YP_123314.1; YP_062 161.1; NP_693109.1; CAB52837.1; ZP_003 33957.1; AAP97897; .1; AAT40119.1; YP_1 49301.1; NP_840123.1; YP_082111.1; 1PJC; AAP07610.1; AAP11530.1; AAM12899.1; YP_021515.1; NP_391071.1; NP_661601.1; YP_081348.1; ZP_00170826.2; ZP_00601 871.1; B? C74218.1; ZP_00241359.1; NP_7 69819.1; ZP_00551096.1; ZP_00462872.1; ZP_00456225.1; YP_174267.1; NP_975075 .1; XP_517776.1; ZP_00375402.1; ZP_0016 7698.2; YP 075651.1; YP_253131.1; YP_0 56938.1; CAH07118.1; NP_682897.1; BAB 06899.1; AAC98487.1; ZP_00659771.1; ZP _00411612.1; AAF11449.1; AAC23577.1; BAC39793.1; ZP_00151223.1; YP_148605. 1; P 17556; YP_041174.1; YP_005051.1; NP _856449.1; YP_186592.1; NP_372233.1; N P_374819.1; YP_144713.1; ZP_00215625.1; ZP_00378064.1; CAE21637.1; ZP_003233 50.1; EAM27747.1; ZP_00497694.1; ZP_00 467473.1; NP_764939. 1; AAK25536.1; ZP_ 00303801.1; YP_159073.1; ZP_00517716.1; ZP_00553800.1; ZP_00629472.1; CAF242 10.1; ZP_00208007.1; ZP_00671129.1; ZP_00008120.2; YP_129373.1; NP_621858.1; NP_470950.1; YP_130399.1; YP_1 11103.1; ZP_00310735.1; AAQ59694.1 AAP95053.1; NP_465104.1; ZP_00400392.1; YP_0141 99.1; AA076661.1; ZP_00231205.1; AAP4 4334.1; YP_005739.1; YP_143482.1; NP_7 97482.1; BAB74054.1; ZP_00120255.1; AA O90629.1; ZP_00636037.1; NP_988633.1; CAB60094.1; CAB59281.2; YP_204286.1; EAM23962.1; YP_040853.1; NP_926915.1; YP_186323.1; C? G43158.1; ZP_0067330 2.1; ZP_00418709.1; P17557; YP_173042.1; ZP_00640011.1; CAD72056.1; CAC46203 .1; ZP_00O49286.2; AAM35807.1; XP_672 369.1; AAC23579.1; AAC23578.1; ZP_001 64800.1; ZP_00507921.1; AA011283.1; YP _266230.1; ZP_00559586.1; ZP_00601825. 1; ZP_00528415.1; CAG35273.1; NP_1021 73.1; NP_085655.1; YP_015797.1; CAG352 69.1; ZP_00621640.1; AAL87460.1; ZP_00 112172.1; ZP_00413882.1; ZP_00130164.2; EAN26936.1; ZP_00579039.1; ZP_005197 76.1; AAZ24151.1; YP_113082.1; ZP_0053 4863.1; ZP_00588083.1; AAK38118.1; ZP_ 00667695.1; NP_969291.1; ZP_00271173. 1; ZP_00513142.1; EAN05956.1; ZP_005321 01.1; AAQ00644.1; ZP_00397135.1; YP_ 15 5059.1; NP_440110.1; NP_768378.1; YP_0 09793.1; AAR37813.1; AAV93547.1; NP_9 53341.1; N87044.1; AAK99657.1; AAS 52072.1; ZP_00545593.1; ZP_00533303.1; NP_636464.1; YP_244224.1; E? M73436.1; ZP_00526469.1; C? J06319.1; NP_953750. 1; CAG36161.1; ZP_00528447.1; ZP_00051 957.1; ZP_00589100.1; ZP_00053708.2; ZP _00570829.1; ZP_00571033.1; ZP_005470 1; ZP_00566160.1; YP_2666937l; YP_1146 39.1; EAN06465.1; ZP_00658424.1; ZP_00 554273.1; ?? V96338.1; ZP_0031919l.l;? L94166.1; ZP_00045954.1; NP_965043.1; ZP X) 387414.1; ZP_O0379883.1; YP_194 023.1; CAH07950.1; NP_229563.1; ZP_001 43839.1; YP_134776.1; AA075844.1; NP_9 70636.1; YP_054766.1 AAK20249.1; AAQ 66392.1; AAK20247.1; AAK20248.1; YP_0 05676.1; C? C12596.1; NP_465401.1; NP_1 10608.1; AAK20246.1; ZP_00602143.1; CA J06565.1; EAN84686.1; XP_563307.1; EA M94020.1; YP_205176.1; ZP_00585967.1; BAD38226.1; AAP55207.1; NP_716196.1; NP_936778.1; YP_130938.1; ZP_OO356O80 .1; AAF96515.1; A? M10111.1; ZP_005813 53.1; YP_024022.1; AAL67502.1; ?? L675 04.1; AAL67501.1; NP_800344.1; AAL675 06.1; AAL67503 .1; ZP_00209144.1; AAL6 7500.1; BAB96979.1; BAB96978.1; AAL67 505.1; B AB97060.1; BAB96906.1; BAB970 12.1; B? B97100.1; B? B97118.1; BAB9690 8.1; BAB97072.1; BAB97009.1; BAB97061 .1; BAB97052.1; BAB97051.1; BAB96990. 1; BAB97098.1; CAA58847.1; BAB96987.1; BAB97161.1; BAB97034.1; B? B96918.1; BAB96980.1; B AB97031.1; BAB96887.1; BAB96882.1; B AB97066.1; B AB97152.1; BAB96968. BAB96859.1 BAB96821.1 BAB97093.; BAB97026.1 BAB97015.1 BAB96860. ?? K20256.1 B? B97110.1 B? B96913. C? D39284.1 B? B97046.1 BAB96985. AAK20257.1 YP 81413.1; B? B97156. BAB96953.1 CAD39277.1 BAB97140. BAB97068.1 B? B96976.1 BAB97064. BAB97145.1 B? B97132.1 BAB97027. BAB97013.1 BAB97113.1 B? B97048. BAB97024.1 BAB96920.1 CAD39252. BAB97091.1 BAB96984.1 BAB96927. BAB96865.1 BAB97018.1 BAB96954. BAB97106.1 B? B97041.1 BAB97021. B? B96938.1 BAB97025.1 BAB97049. BAB97005.1 B? B96910.1 BAB97008. BAB97148.1 BAB96988.1 BAB96951. BAB96941.1 BAB97155.1 BAB96993. BAB96904.1 B? B96875.1 BAB97144. BAB97006.1 BAB96956.1 BAB96924. BAB97108.1 BAB96930.1 AAK20250. BAB97162.1 BAB96872.1 B? B96858. C? D39278.1 NP 078054.1 BAB97159. BAB96952.1 BAB96948.1 BAB97117. BAB97070.1 BAB97104.1 BAB97103. BAB97083.1 B? B97055.1 BAB96830. BAB97089.1 AAK20254.1 BAB97139 B? B97053.1 BAB96997.1 BAB96869 BAB96837.1 B? B96925.1 BAB96921. BAB96903.1; BAB96900.1 B? B97028. BAB97136.1; B? B97126.1 BAB97092. ? AK2O258.1; B? B97069.1 ?? K20255 B? B96961.1; BAB96957.1 CAD39216 AAK20253.1; AA037738.1 BAB96977. BAB96888.1; B? B96893.1 CAD39221 AAK20261.1; AAK20251.1 BAB97124 BAB97029.1; B AB96940.1 BAB96876 BAB96842.1; BAB96831.1 C? D39245. AAK20252.1 ¡CAD39266.1 BAB97134. BAB97130.1; BAB97099.1 B? B97032. B? B96848.1; BAB97017.1 BAB97010. B? B96885.1; ?? K20268.1 BAB97123. BAB97063.1; BAB96971.1 BAB97114. BAB97094.1; BAB97056.1 BAB96886. BAB96843.1; BAB97138.1 BAB97135. BAB97016.1; BAB96998.1 BAB96936. BAB96919.1; BAB97160.1 BAB97131 BAB97039.1; BAB97033.1 BAB97023 BAB97003.1; BAB96983.1 BAB96850. BAB96849.1; BAB97030.1 BAB96991. BAB96894.1; B? B96863.1 CAD39217. BAB97133.1; BAB97112.1 BAB971 U. BAB97062.1; BAB97045.1 BAB96929. BAB96874.1; BAB96827.1 BAB97128. BAB97121.1; B? B97096.1 BAB97058. B? B97054.1; BAB97047.1 BAB96970. BAB96922.1; BAB96828.1 CAD39229.1 BAB97078. BAB97011.1 BAB96960.1 BAB96939. B? B97154.1 BAB97142.1 BAB96937. C? D39275.1 BAB97057.1 BAB97037. B? B96992.1 BAB96964.1 BAB96846. B? B96838.1 BAB96829.1 BAB97119. BAB96931.1 AA037727.1 BAB96963. BAB96890.1 BAB96857.1 BAB96844. BAB97129.1 BAB97127.1 BAB97075. BAB96975.1 BAB96899.1 BAB96889. BAB96881.1 CAD39224.1 BAB97149. BAB97073.1 BAB97043.1 BAB97038. CAD39240.1 B? B97086.1 B? B97004. C? D39283.1 CAD39228.1 CAD39222. CAD39220.1 AAK20260.1 BAB97088. CAD39272.1 BAB97137.1 BAB97059. BAB96965.1 BAB96911.1 BAB96905. BAB96841.1 CAD39237.1 BAB97014.; BAB96898.1 BAB96996.1 BAB96835. BAB97084.1 BAB96867.1 BAB96822. BAB97125.1 BAB9710J1 BAB96907. CAD39249.1 C? D39236.1 C? D39213. C? D39212.1 B? B97105.1 BAB96902. C? D39279.1 BAB97116.1 BAB96866. B? B97102.1 AAK20259.1 BAB97040. CAD39256.1 BAB97157.1 BAB97071. BAB96972.1 CAD39261.1 AA037723. BAB97101.1 BAB97095.1 BAB96879. CAD39248.1 CAD39231.1 CAG36956.1; BAB96955.1; BAB96943.1 CAD39233.1; BAB97081.1; BAB97150.1 B? B96999.1; B? B96949.1; B? B96854.1 B? B96947.1; C? D39263.1; AAK20269.1 BAB96967.1; BAB96962.1; BAB96917.1 CAD39264.1; AA037741.1; BAB96873.1 BAB96839.1; CAD39269.1; CAD39234.1 BAB97151.1; BAB97085.1; BAB97080.1 B AB96995.1; CAD39280.1; BAB97153.1 CAD39242.1; CAD39215.1; AAO37740.1 BAB96891.1; CAD39243.1; BAB97079.1 BAB97042.1; BAB96840.1; C? D39241.1 C? D39274.1; ?? K20262.1; BAB97050.1 CAD39271.1; CAD39265.1; AA037743.1 AA037729.1; BAB96923.1; BAB96884.1 BAB96825.1; CAD39251.1; AA037735.1 CAD39282.1; BAB96861.1; AA037724.1 CAD39270.1; CAD39235.1; BAB96933.1 B AB96946.1; B AB97146.1; BAB96986.1 BAB96896.1; CAD39232.1; AA037736.1 B? B96959.1; ?? K20267.1; CAD39223.1 C? D39244.1; B? B96966.1; BAB97141.1 CAD39276.1; BAB96935.1; CAD39255.1 CAD39209.1; BAB97115.1; CAD39253.1 A? O37730.l; B? B96926.1; BAB96870.1 AA037737.1; BAB96958.1; BAB97022.1 CAD39250.1 Iformil cvonvierte NP 600108 NP 737565.1 ZP_00655917.1; NP_939225 tetrahydrofolate formyl-THF .1; BAC71386.1; CAB59679.1; ZP_002926 cyclo ligase at 5.10-66.1; YPJ 21170.1; YP_061656.1; ZP_0054 (EC 6.3.3.2.) Methenyl- 5597.1; NP_301256.1; NP_959857.1; ZP_00 (R76) THF 573419.1; C? I37699.1; NP_214187.1; SO U; YP_075021.1; NP_696710.1; ZP_004122 60.1; ZP_00134730.1; EAM72866.1; ZP) 0 575597.1;? A089633.1; NP_215507.1; NP_ 854676.1; AA 45268.1; EAN27919.1; ZP_ 00552794.1; NP_882562.1; ZP_00326738.1; YP_115170.1; NP_886754.1; NP_881632. 1; NP_534225.1; NP_440379.1; ZP_006267 22.1; ZP_00600182.1; YP_079816.1; YP 4 8302.1; ZP_00519228.1; NP_439018.1;? A 044764.1; NP_266325.1; ZP_00323162.1; ZP_00156713.1; ZP_00110015.2; ZP_0047 1373.1; ZP_00534935.1; ZP_00159040.2; Z P_00171408.2; ZP_00398486.1; YP_19066 5.1; ZP_00396530.1; YP_009590.1; AAQ66 843.1; YP_125990.1; AAR37522.1; YP_270 316.1; YP_094629.1; YPJ 22981.1; AAL00 707.1; NP_930816.1; BAB82030.1; ZP_001 55857.2; YP_222388.1; ZP_00269658.1;? AX87904.1; AAL51496.1; YP_233427.1; C AG35375.1; AAK79064.1; BAB76524.1; N P_794958.1; AAN70768.1; AAO09973.1; A A079363.1; ZP_00585069.1; AAN48446.1; ZP_00538598.1; ZP_00417043.1; EA0243 95.1; ZP_00638539.1; AAQ58176.1; ZP_00 403498. 1; YP_144877.1; AAX66908.1; CA C41462.1; NP_952189.1; YP_055204.1; ZP _00347044.1; YP_005216.1; 1 WKC; YP_15 6483.1; E? O17392.1; YP_175218.1; ZP_00 594221.1; ZP_00319181.1; AAH19921.1; N P_0O10O9349.1; NP_798970.1; ZP_005567 64.1; NP_253915.1; ZP_00630769.1; AAT4 9915.1; ZP_00141705.1; ZP_00264800.1; N P_081105.1; CAC47337.1; ZP_00055330.1; AAT42396. 1; NP_102175.1; CAD13617.1; ZP_00062893.1; ZP_00315438.1; AAF956 21.1; NP_971601.1; ZP_00588425.1; YP_03 4143.1; NP_923705.1; C? H06623.1; YP_09 8246.1; NP_670599.1; NP_311809.2; ZP_00 630026.1; YP_263029.1; AAG10441.1; ZP_00592340.1; YP_170963.1; YP_131238.1; NP_614888.1; CAHO3038.1; AAL95098.1; BAA28715.1; EAN08520.1; YP_152082.1; NP_726312.1; NP_611785.1; NP_726311.2 ; YP_071689.1; CAF26595.1; NP_755368.1; NP_681770.1; AAR38106.1; NP_840158.1; ZP_00509367.1; H 12417.1; NP_10494 3.1; ZP_00634551.1; EAA26463.1; ZP_005 82514.1; AAN58080 .1; NP_768167.1; ZP_0 0211724.1; NP_638609.1; NP_965424.1; X P_308920.2; NP_785167.1; ZP_00004961.1 ; AAK25207.1; AAF 11368.1; BAB05136.1; ZP_00153791.1; NP_949790.1; EAM47244 .1; ZP_00380928.1; YP_205487.1; ZP_0024 3180.1; ZP_00152677.2; ZP_00623093.1; Z P_00529088.1; ZP_00511292.1; ZP_00655 348.1; NP_975489.1; NP_806671.1; NP_77 7982.1 P_00388995.1; XP_413857.1; ZP_ 00123619.1; AAC75949. 1; NP_708674.2; A AV63351.1; YP_067406.1; A? K03805.1; A AH89273.1; AAG58038.1; YP_253281.1; X P_478853.1; NP_360331.1; YP_021133.1; Z P_00530394.1; NP_220841.1; NP_470709 1; ZP_00046926.1; ZP_00334623.1; ZP_001 43346.1; ZP_00376747.1; EAN07877.1; NP _464861.1; B31354.1; YP_013951.1; NP _692845.1; CAD74215.1; ZP_00278951.1; NP_716405.1; NP_980638.1; EAM31460.1; ZP_00463106.1; AAL21936.1; CAE07229. 1; CAC05433.1; AAM60972.1; AA041971. 1; YP_085596.1; AAM90961.1; AAV96195 , 1; XP_510537.1; NP_66O736.1; CAB83635 .1; YP_246824.1; AAC65661.1; NP_764791, 1; NP_878554.1; CAG86858.1; ZP_003868 75.1; YP_194357.1; W90586.1; ZP_004 34550.1; ZP_00492814.1; YP_053491.1; C AG73377.1; AA035597.1; EAL2516J1; ZP _00499196.1; CAG80466.1; AAW70845.1; YP_045820.1; NP_734906.1; AAM99309.1; YP_038325.1; CAB66452.1; AAV48062.1; AAH24567.1; YP 041023.1; YP 186447.1 ; NP_372074.1; ZP_00385074.1; NP_39036 9.1; XP_591041.1; XP_588513.1; AAV8883 9.1;? AF42416.1; ?? P11177.1; ZP_002385 35.1; ?? M38252.1; YP_153863.1; ?? Z195 06.1; YP_266014.1; NP_968104.1; NP_816 417.1; YP_255152.1; C? A12119.1; CAH89 819.1; BAB14383.1; BAC43679.1; BAB147 39.1; CAA21728.1; 1 YDM; ZP_00544875.1; NP_766349.1; NP_779018.1;? AB84710.1; ZP_00210813.1; ZP_00579599.1; YPJ 70 178.1; AAX78148.1; NPJ565139.1; YPJ99 768.1; AAF73519.1; ZP_0O3O3053.1; XP_5 11153.1; U37082.1; NP_280554.1; EAK 93783.1; AAH79691.1; AAS51535.1; AAH 94085.1; YP J 58939.1; NP_966978.1; CAF 99528.1; ZP_0O370459.1; ZPJ) 0652045.1; ZP_00548066.1; ZP_00660458.1; XP 958 64.2; NP_341992.1; CAF 18476.1; NP_2992 95.1; C? A83541.1; B? B65129.1; ZP_0037 3105.1; YPJ80354.1; NP_220167.1; XP_4 27228.1; EAL28752.1; NP_011110.1; XP_6 94566.1; XP_414185.1; C? E66846.1; ZPJ) 0310271.1; CAE21313.1; C? H64402.1; XP J545889.1; XP J > 98686.1; A? H37852.1; E AN86573.1; AAP98720.1; ZP W367232.1; ZP_00O63393.2; NP_661939.1; AAP05733. 1; NP_701792.1; XP_759454.1; AAL64263. 1; EAN86747.1; AAF52130.1; XP 10457. 1; YP_076510.1; ZP_00160457.2; NP_2286 90.1; YP_242180.1; YP > 69261.1; NP_638 415.1; NP_797008.1; ZP_00208451.1; ?? L 76402.1; ZP_00595723.1; ZP_00512971.1; CAE10117.1; ZP_00242385.1; ZP_005365 77.1; AAR38305.1; NP_948106.1; ZP_0039 8593.1; ZP_00654525.1; ZP_00531189.1; N P_946915.1; NP_930734.1; NP_661504.1; Z P_00005143.1; NP_841729. 1; C AE21050.1; CAE07366.1; YP_047868.1; CAD78524.1; ZP_00283981.1; NP 767875.1; NP 349925 .1; YP_221536.1; AAL52347.1; AAP99844. 1; ?? N29722.1; ZP_00589323.1; ZP_00624 467.1; CAG81467.1; ZP_00562640.1; EAN 27962.1; ZP_00309879.1; ZP_00556008.1; YPJ 71853.1; ZP_00170753.1; YPJ 60629.l; E? O18327.1; YPJ56395.1; ZP_001511 82.1; ZP_00395964.1; CAG37235.1; ZP_00 410921.1; ZP 00269047.1; AAM06094.1; N P_772921.1; NP_892760.1; CAG73733.1; N P_766885.1; EAL85880.1; XP_761728.1; A? O77030.1;? A61543.1; ZP_00169873.1; ZP_00638540.1; ZP_00527619.1; AAV947 18.1; EAM43226.1; ZP_00661517.1; CAD1 5264.1; P_00268814.1; ZP_00620303.1; Z P_00634556.1; AAF10450.1; EAA59015.1; AAV31651.1; NP_953471.1; NP_771607.1; EAA67392.1; AAQ59608.1; EA 75775.1; NP_9495877l; ZP_00595151.1; NP_716721 .1; ZP_00534594.1; ZP_00305184.1; ZP_00 592653.1; AAK02822.1; ZP_00269954.1; Z P_00543698.1; ZP_00552392.1; C? H0904 3.1; EAA56840.1; CAA22286; 1; ZP_00650 573.1; CAC45827.1; NPJ 08558.1; NP_531 944.1; YP_004383.1; YP J 44026.1; YP_26 6440.1; NP_693560.1; YP_257642.1; ZP_00 134047.2; ZP_00581547.1; CAB99179.2; Y PJ 59786.1; EAM73042; .1; CAG36429.1; A A035727.1; YP_022333.1; ZP_00239222.1 ; ZP_00269964.1; YP_086673.1; NP_98182 7.1; ?? P12268.1; ?? P77233.1; YP_03939 7.1; ZP_00208374.1; E? N06205.1; CAE109 04.1; AAK99898.1; YPJ 79865.1; ZP_0038 7486.1; CAH00339.1; CAB73713.1; CAC34 631.1; ZP_00370704.1; NP_013406.1; AAV 47483.1; ZP_00489181.1; AAS51890.1; NP _947707.1; EA 94256.1; CAG88952.1;? A F01454.1; A? F01453.1; NP_281028.1; CA G58594.1; EAM46849.1; AAV47781.1; ZP _00566262.1; ZP_00402706.1; YP_234755 1; ZP_00334304.1; YP_259190.1; NP_7935 84.1; EAO 18747.1; ZP_00473306.1; AA03 6990.1; NP_840779.1 ZP 00333201.1; ZP_ 00265576.1; NP_251797.1; ZP_00535298.1; ZP_00204956.1; 1 Y4I.AAC83351.1; AAO 46884.1; A? N32868.1; YP_074661.1; B? C 02724.1; ZP_00348658.1; ZP_00593187.1; NP_947695.1; AAV68695.1; AAK29460.1; ZP_00168139.2; YPJ 14900.1; A? L95612. 1; NP_972801.1; YP_046919.1; YP_200029. I; ZP_00317087.1 ZP 00280956.1; ZP) 0 416859.1; YPJ 11696.1; ZP_00502001.1; Z P_00466954.1; AAR38140.1; ZP_0044846 0.1; NP_950100.1; ZP_00395704.1; ZP J) 04 57013.1; NP_717420.1; ZP_00464307.1; A? Q65554.1; ZP_00213081.1; AAQ60395.1; YP J 59732.1; ZP_00637444.1; ZP_00425 258.1; ZP_00643937.1; BAB55900.1; ZP_0 0048836.2; NP_767745.1; YP_268443.1; ZP _00583389.1; ZP_00396707.1; ZP_006361 23.1; AAV94639.1; EAN05079.1; AAF 1173 0.1; NP_623174.1; ZP_00587584.1; AAZ18 645.1; CAE10114.1; ZP_00303125.1; ZP_0 0623125.1;? AK78087.1; ZP_00655219.1; CAA09983.1; ZP_00624529.1; AAK24209. 1; ?? L52798.1; AAB03240.1; ZP_0054251 8.1; NP _531136.1; AAK25130.1; AAN6693 2.1; P13254; ZP_00400550.1; B? B04518.1; YP_221092.1; NP_945724.1; NP_970501.1; AAU37954.1; NP_981084.1; 1 GC2; ZP_00 548195.1; NP_635109.1; CAA04124.1; ZP_ 00236112.1; YP_004765.1; ZP_00055526.1 ; ZP_00207149.1; A? M05915.1; YP_08597 6.1; 1E5F; ZP_00308639.1; ZP_00578607.1 Media and Culture Conditions for the Growth of Escherichia coli and Corynebacterium glutamicum The person skilled in the art is familiar with the culture of common microorganisms such as C. gl utami cum and E. coli. In this way, a general education will be given in the following regarding the cultivation of C. gl utamicum. The corresponding information can be retrieved from standard textbooks for the cultivation of E. coli E. coli strains are routinely grown in MB and LB broth, respectively (Follettie, MT, Peoples, 0., Agoropoulou, C, and Sins ey, A J. (1993) J. Bacteriol., 175, 4096-4103 ). Minimum means for E. coli are M9 and modified MCGC (Yoshihama, M., Higashiro, K., Rao, EA, Ando, M., Shanabruch, W G., Follettie, MT, Walker, G. C, and Sinskey, AJ (1985) J. Bateriol., 162, 591-507), respectively. Glucose can be added to a final concentration of 1%. Antibiotics may be added in the following amounts (micrograms per milliliter): ampicillin, 50; Kanamycin, 25; Nalidixic acid, 25. Amino acids, vitamins, and other supplements can be added in the following amounts: methionine, 9.3 mM; arginine, 9.3 mM; histidine, 9.3 mM; thiamin, 0.05 mM. The cells of E. Coli are grown routinely at 37 C, respectively. Genetically modified Corynebacteria are typically grown in synthetic or natural growth media. A number of different growth media for Corynebacteria are well known and readily available (Lieb et al. (1989) Appl. Microbiol. Biotechnol., 32: 205-210; von der Osten et al. (1998) Biotechnology Letters , 11: 11-16, Patent DE 4,120,867, Liebl (1992) "The genus Corynebacterium, in: The Procaryotes, Volume II, Balows, A. et al., Eds. Springer-Verlag.) These media consist of one or more carbon sources, nitrogen sources, inorganic salts, vitamins and trace elements Preferred carbon sources are sugars, such as mono, di, or polysaccharides, eg, glucose, fructose, mannose, galactose, ribose, sorbose, ribose, lactose , maltose, sucrose, raffinose, starch or cellulose serve as very good sources of carbon.It is also possible to supply sugar to the media by means of complex compounds such as molasses or other byproducts of sugar refining. carbon sources. Other possible sources of carbon are alcohols and organic acids, such as methanol, ethanol, acetic acid or lactic acid. Nitrogen sources are usually organic or inorganic nitrogen compounds, or materials that contain these compounds. Exemplary sources of nitrogen include ammonia gas or ammonia salts, such as NH4C1 or (NH) 2S04, NH40H, nitrates, urea, amino acids or complex sources of nitrogen such as corn liquor, soybean meal, soybean protein, yeast extract, meat extract and others. The overproduction of methionine is possible using different sources of sulfur. Sulfates, thiosulfates, sulphites and also smaller sources of sulfur such as H2S and sulfides and derivatives can be used. Also organic sources of sulfur such as methyl mercaptan, thioglycolates, thiocyanates, and thiourea, sulfur-containing amino acids such as cysteine and other sulfur-containing compounds can be used to achieve efficient production of methionine. Formate may also be possible as a supplement as are other sources of Cl such as methanol or formaldehyde,). Particularly suitable are methanethiol and its dimethyl disulfide dimer. Compounds of inorganic salts that may be included in the media include the chloride, phosphorous or sulfate salts of calcium, magnesium, sodium, cobalt, molybdenum, potassium, manganese, zinc, copper and iron. Chelating compounds can be added to the medium to keep the metal ions in solution. Particularly useful chelating compounds include dihydroxyphenols, such as catechol or protocatequate, or organic acids, such as citric acid. It is typical for the media to also contain other growth factors, such as vitamins or growth promoters, examples of which include biotin, riboflavin, thiamine, folic acid, nicotinic acid, pantothenate and pyridoxine. Factors and growth salts frequently originate from components of complex media such as yeast extract, molasses, corn liquor and others. The exact composition of the media compounds depends strongly on the immediate experiment and is decided individually for each specific case. Information about media optimization is available in the textbook "Applied Microbiol. Physiology, A Practical Approach (Eds. PM Rhodes, PF Stanbury, IRL Press (1997) pp. 53-73, ISBN 0 19 963577 3) It is also possible to select growth media from commercial suppliers, such as standard 1 (Merck) or BHI (grain-heart infusion, DIFCO), etc. All the components of the medium must be sterilized, either by heat (20 minutes at 1.53 kgf / cm2 (1.5 bar) and 121 C) or by sterile filtration. The components can be sterilized together or, if necessary, separately. All media components can be presented at the start of growth, or optionally can be added continuously or in batches. The culture conditions are defined separately for each experiment. The temperature should be in a range between 15 ° C and 45 ° C. The temperature can be kept constant or can be altered during the course of the experiment. The pH of the medium can be in the range of 5 to 8.5, preferably around 7.0, and can be maintained by the addition of buffers to the media. An exemplary buffer for this purpose is a potassium phosphate buffer. Synthetic tampons such as MOPS, HEPES, ACES and others can be used alternatively or simultaneously. It is also possible to maintain a constant culture pH through the addition of NaOH or NH 4 OH during the course of growth. If components of complex medium such as yeast extract are used, the need for additional buffers can be reduced, due to the fact that many complex compounds have high buffering capabilities. If a fermenter is used to grow the microorganisms, the pH can also be controlled using gaseous ammonia. The incubation time is usually in a range of several hours to several days. This time is selected in order to allow the maximum amount of product to accumulate in the broth. The growth experiments described can be carried out in a variety of containers, such as microtiter plates, glass tubes, glass flasks or glass or metal fermenters of different sizes. To select a large number of clones, microorganisms must be cultured in microtitre plates, glass tubes or shake flasks, with or without baffles. Preferably 100 ml shake flasks, filled with 10% (by volume) of the growth medium required, are used. The flasks should be shaken on a rotary shaker (25 mm amplitude) using a speed range of 100-300 rpm. Evaporation losses can be reduced by maintaining a humid atmosphere; alternatively, a mathematical correction for evaporation losses must be made. If genetically modified clones are tested, an unmodified control clone or a control clone containing the basic plasmid without any insert should also be tested. The medium is inoculated to an OD600 of 0.5-1.5 using cells grown on agar plates, such as CM plates (10g / l of glucose, 2.5g / l of NaCl, 2g / l of urea, 10g / l of polypeptone, 5g / l of yeast extract, 5g / l of meat extract, 22g / l of NaCl, 2g / l l of urea, 10g / l of polypeptone, 5g / l of yeast extract, 5g / l of meat extract, 22g / l of agar, pH 6.8 with 2M NaOH) that has been incubated at 30 C. Inoculation of the media is achieved by the introduction of a saline suspension of C. gl utami cells from CM plates or addition of a liquid preculture of this bacterium. The invention will now be illustrated by means of various examples. These examples, however, are not intended in any way to limit the invention in any way.

Examples The modalities within the specification provide an illustration of the modalities in this description and should not be construed to limit their scope. The skilled technician readily recognizes that many other modalities are encompassed by this description. All publications and patents cited and sequences identified by access reference numbers or database in this description are incorporated for reference in their entirety. To the extent the extent to which the material incorporated for reference contradicts or is inconsistent with the present specification, this specification will replace any material. The citation of any references herein is not an admission that such references are prior art to the present disclosure. Unless otherwise indicated, all numbers expressing ingredient quantities, cell culture, treatment conditions, and so forth used in the specification, including claims, shall be understood as modified in all cases by the term "about . " Accordingly, unless otherwise indicated, the numerical parameters are approximations and may vary depending on the desired properties sought by the present invention. Unless stated otherwise, the term "at least" preceding a series of elements should be understood to refer to every element in the series. Those skilled in the art will recognize, or be able to ascertain using no more than routine experimentation, many equivalents to the specific embodiments of the invention described herein. Such equivalents are intended to be encompassed by the following claims A) Optimal metabolic flow prediction for an organism with increased efficiency of methionine synthesis Constructing the metabolic networks for C. gl utami cum and E. coli Red of C. gl utami cum. The basic metabolic network of the wild type of C. glutamicum was established for the use of glucose and sulphate as a source of carbon and sulfur, respectively (http://www.genome.jp/kegg/metabolism.html). It includes glucose uptake by a phosphotransferase system (PTS), glycolysis (EMP), pentose phosphate pathway (PPP), tricarboxylic acid cycle (TCA), anaplerosis and respiratory chain. The assimilation of sulfate comprises uptake and subsequent conversion to hydrogen sulfide (Schiff (1979), Ciba Found Symp, 72, 49-69). In the stoichiometric model, the sulphate assimilation route was concentrated in 2 reactions: the reduction of sulphate to sulfite that requires 2 ATP and 1 NADPH and the reduction of sulfite to sulfur that requires 3 NADPH. The complete model consisted of 59 internal and 8 external metabolites. The external metabolites comprise substrates (glucose, sulfate, ammonia, oxygen) and products (biomass, C02, methionine, glycine). Glycine was considered as an external metabolite, since once formed as a by-product it can not be re-used by C. gl utami cum (http://www.genome.jp/kegg/metabolism.html). In total, the metabolic network contains 62 metabolic reactions, of which 19 were considered reversible. For ATP production in the respiratory chain, a P / 0 ratio of 2 (for NADH) and 1 (for FADH) was assumed (Klapa et al. (2003) Eur. J. Biochem., 27017, 3525-3542) .. The demand for precursors for biomass formation was taken from the literature (Marx et al. (1996) Biotechnol. Bioeng, 49 (2), 111-129). The demand for sulphate and ammonia for the biomass was calculated from the content of the different amino acids in the biomass. The model for C. glutamicum is shown in Figure 1. Network of E. coli The construction of the model for the central metabolism of wild type E. coli was based on the literature (Carlson et al. (2004), Biotechnol. Bioeng., 851, 1-19), and databases (http: / /www.genome.jp/kegg/metabolism.html). The model for growth and production of methionine in glucose and sulfate comprised glucose uptake by PTS, EMP, PPP, TCA cycle, anaplerosis, respiratory chain and sulfate assimilation. The metabolic network contained 64 metabolic reactions, so 20 were considered reversible. Glucose, sulfate, ammonia and oxygen were considered as external substrates, biomass, C02 and methionine as external products. For interconversion of NADH and NADPH, a reversible transhydrogenase was considered (Yamaguchi et al (1995), J. Biol. Chem., 27028, 166653-9). Moreover, it was considered that E. active coli homoserin by succinylation instead of acetylation (R40) (Sekowska et al. (2000), J. Mol.Micorbiol.Biotechnol, 22, 145-177). Additionally, a glycine cleavage system (R71, R72) was considered. For ATP production in the respiratory chain, P / O ratios of 2 (for NADH) and 1 (for FADH) were assumed (Carlson et al. (2004), vide supra). The demand for precursors for biomass formation was taken from the literature (Edwards et al. (2000); Weber et al. , (2002). Modifications of networks. In additional simulations the stoichiometric networks described in the above were modified. This involved the elimination or insertion of different reactions and pathways potentially of interest to improve the production of methionine. Additionally, carbon and sulfur sources were varied to investigate their influence on methionine production. Analysis of metabolic pathways. The analysis of metabolic pathways was carried out using METATOOL (Pfeiffer et al., (1999), Bioinforma ti cs, 153, 251-7, Schuster et al. (1999) Trends Biotechnol, 172, 53-60). The version used (meta 4 .0 l_double. Exe) is available on the internet http: // www. biozentrum. uni-wuerzburg from / bioinformatik / computing / metatool / -pinguin. biologie.uni-jena from / bioinformatik / networks /). The mathematical details of the algorithm are described in Pfeiffer et al. (vide supra) which is incorporated by this for reference with respect to the manner in which METATOOL software should be used. The analysis of metabolic pathways resulted in several hundred modes of elementary flow for each situation investigated. For each of these flow modes, the carbon yields of the biomass (Y? / S) and methionine (YMet / s) were calculated as a percentage of the carbon that entered the system as a substrate. Throughout the work it is given in percentage values ((C-mol) (C-mol substrate) "1 x 100. Therefore also the co-substrates, such as formate or methanethiol and its dimethyldisulfide dimer were considered. The comparative analysis of all the elementary modes obtained for a certain network scenario allowed the determination of the theoretical maximum yields Y? / S, ma and ^ Met / S, max- Resulted and Implications of the model Comparison of methionine production by C. gl utami cum and E. coli The two most promising organisms for biotechnological production of methionine are C. gl utami cum and E. coli To evaluate the potential of these two organisms, the analysis of metabolic pathways was carried out as described in the above. Initially wild type networks were investigated. As shown for the wild type of C. gl utami cum and E. coli, a large number of elementary flow modes with different carbon yields were obtained for biomass and methionine (Figures 2 A, B). Among the modes observed, most are extreme modes exclusively linked to the production of biomass or methionine. These are given in the two axes of the graph. In addition, flow modes were also produced with simultaneous production of biomass and methionine. The maximum theoretical yield in biomass was 88.5% for C. gl utamicum and thus slightly lower than as found for E. coli with 91.7%. Both organisms have a high potential to produce methionine. The maximum theoretical carbon yield for methionine of C. glutami cum was 48.6% (Figure 2A). E. coli shows a significantly higher value of 56.2% (Figure 2 B). The superior potential of the wild type of E. coli may indicate favorable characteristics of your metabolic network. This aspect was studied in additional simulations (see below). Closer inspection points to two reactions, namely, the glycine cleavage system and transhydrogenase, which may be beneficial for increased methionine production. In fact the optimal solution found for C. glutami cum wild type binds to the substantial formation of glycine, which can not be re-used, while glycine does not accumulate for an optimal production of methionine by E. wild type coli. With respect to the high demand of 8 NADPH for methionine, also the availability of transhydrogenase for interconversion of NADH and NADPH in E. coli may contribute to the observed superior efficiency To further investigate the importance of these reactions for methionine production, additional simulations were carried out assuming different genetic modifications of the underlying metabolic networks (see below).

Metabolic fluxes in C. glutamicum and E. coli under conditions of optimal methionine production First, the metabolic networks of both organisms were studied in greater detail to identify which of the available pathways are involved in optimal production of methionine and which pathways should be dispensable . For this purpose, the metabolic flux distribution was calculated for the optimal elementary modes of C. gl utamicum and E. col i, that is, the mode with the highest theoretical methionine yield. This is why all flows are given as relative molar values, normalized to the rate of glucose uptake, as is usually done in metabolic flux analysis. Note that the flows (given in mol (mol) -1 x 100) differ from the carbon yields (in C-mol (C-mol) "1 x 100) used to describe the maximum performance. Basic models (Figure 1) that were inactive in the respective modes were erased from Figures 3 and 4. The flow distribution for optimal production of methionine in the two organisms differed dramatically (Figures 3, 4) .The optimal flow towards methionine in C. gl utami cum it was 58.3% For this purpose, C. gl utamicum exhibited a very high PPP activity with a flow through the oxidative reactions of the PPP of 250% This is probably due to the demand of NADPH since 8 NADPH must be supplied for methionine synthesis, mainly for sulfur reduction, the flow to the PPP is substantially higher than the glucose uptake flow Glucose 6-phosphate isomerase, which works in the gluconeogenic direction, also contributes significantly atively supplying carbon to the PPP. The TCA cycle is completely switched off, so that isocitrate dehydrogenase does not contribute to the formation of NADPH. Additionally, C. gl u tamicum employs two important metabolic cycles. The first cycle only involves 2-oxoglutarate and glutamate, which interconvert at high flow, to assimilate ammonium and use it for required amination reactions. These are the formation of methionine itself and the formation of serine as a donor of the methyl group for methyl-THF formation, so that the flow through this cycle is exactly twice the flow of methionine. The second metabolic cycle comprises the meetings of pyruvate, oxaloacetate and malate. It exhibits two main functions: Almost half of the C02 lost in the oxidative PPP reentrates in the metabolic network by the highly active C02 fixation (125% flow). Additionally, the combination of the three enzymes involved in the cycle acts as a transhydrogenase and interconverts NADH in NADPH (25% flow). For this reason C. gl utamicum can, to a certain extent, overcome the deficiency of a transhydrogenase.

The optimal production of methionine in E. coli resulted in a methionine flux of 67.5%. In contrast to C. gl utamicum, the PPP was not active, while the TCA cycle showed a high flow of almost 100%. However, the TCA cycle was operating in a modified form. The succinyl-CoA to succinate stage is connected by the corresponding reaction producing succinate in the methionine biosynthesis. Interestingly, optimal production of methionine required substantial activity of the glyoxylate deviation (31% flow). More significant is the enormous flow of 574% through the transhydrogenase from NADH to NADPH. This highlights the importance of this enzyme for an efficient production of methionine in E. coli As shown in the above, the maximum theoretical yield of methionine falls significantly (Figure 2 D), when this enzyme is removed. In both organisms pyruvate kinase is dispensable for optimal production of methionine. Accordingly, the flow of PEP to pyruvate is provided exclusively by the PTS, coupled to the glucose uptake. Obviously, pyruvate kinase is not required for ATP production. The deactivation of this enzyme can be an interesting objective, since it can limit the flow to pyruvate and related excess metabolites of the TCA cycle. In summary, the optimal flow distribution of the two organisms was fundamentally different. By using elemental flow mode analysis with respect to the methionine synthesis, predictions can be obtained for genetic modifications that should allow to increase the efficiency of methionine synthesis.

Potential improvement of methionine production by genetic modifications To study the influence of some key reactions in greater detail, additional simulations with modified metabolic networks were carried out. The implementation of a transhidrogenase in C. gl utami cum led to an increased theoretical methionine yield of 51.9% (Figure 2 C). The deactivation of transhydrogenase in E. coli strongly decreased its potential for methionine production (Figure 2 D). This highlights the beneficial effect of an active interconversion of NADH in NADPH for methionine production. The insertion of the glycine cleavage system in C. gl utamicum increased the theoretical maximum yield of methionine to 56.5% (Figure 2 E). Similarly, deactivation of the glycine cleavage system or the transhydrogenase in E. coli resulted in a reduced maximum theoretical yield of methionine (Figures 2 F, D). Note that the transhydrogenase also affects the maximum theoretical yield in biomass. The insertion in C. gl utami cum leads to an increase, while the elimination in E. coli causes a decrease in Y? / S, ma respectively. Concerning carbon yields, all flow modes were located within a triangle shaped space, which extended between the origin and the two extreme flow modes with maximum biomass and methionine formation, respectively (Figures 2 A-F). The connection between the two extreme modes thus shows an optimal line, which gives the maximum possible methionine yield under different growth regimes. All modes and linear combinations of modes on this line represent interesting solutions for a production process. A real production process will always be linked to some biomass formation.

Influence of the sulfur source on methionine production Potentially positive effects of genetic modifications can be clearly identified. Additional simulations were carried out to even further increase the theoretically possible methionine synthesis efficiency. In this regard, the effect of alternative nutrients was investigated. Because of this, the sulfur source can play a central role. The results obtained are exemplified for C. gl utami cum.

The source of conventional sulfur is sulfate as it is also applied in the analysis of ways in the above for wild types. The assimilation of sulphate, however, joins a high demand of 2 ATP and 4 NADPH. Especially the high requirement of reducing power suggests that the state of reduction of the sulfur source could be a crucial point. Accordingly, an analysis of metabolic pathways was carried out using sulfate, thiosulfate, and sulfur as sources of sulfur. For the use of thiosulfate, thiosulfate reductase was incorporated (Schmidt et al (1984) vide supra, Heinzinger et al (1995) J. Bacteriol, 177: 2813-2820, Fong et al. (1993) J. Bacteriol, 175 : 6368-6371)) in the model. This enzyme allows the cleavage of thiosulfate in sulfite and sulfur and thus reduces the global demand for NADPH for methionine production by about 25%. It should be noted that, as far as is known, the consumption of both sulfur atoms of the thiosulfate has not yet been shown in C. gl utamicum. Another possibility to produce sulfur from a smaller form of sulfur is the so-called anaerobic sulfite reductase (Huang et al (1991) Journal of Bacteriology 173 (4): 1544-53). It becomes obvious that the source of sulfur is a key point concerning the theoretical carbon yield of a production process. In comparison with sulphate (Figure 2 A), the use of alternative sulfur sources significantly increases the maximum theoretical yield (Figures 3 A, B). The increase in sulphate (48.6%) to thiosulfate (57.8%) to sulfur (63.4%) impressively highlights the high potential of using alternative nutrients for methionine production. Additionally, it demonstrates the high importance of reducing power (NADPH) for optimal methionine biosynthesis. In C. glutamicum NADPH is produced mostly in the oxidative PPP and to a certain extent in the TCA cycle with an isocitrate dehydrogenase dependent on NADP. In the course of sulfate growth, the wild type requires 8 moles of NADPH per mole of methionine synthesized by direct sulfhydrylation and 9 moles of NADPH via the transulfurization route (Hwang et al. (2002), J. Bacteriol, 1845, 1277-86). The thiosulfate network requires 6 moles of NADPH for methionine production and thus 25% less. The sulfide-consuming network will only require 50% of the NADPH sulphate demand. This gradual reduction in NADPH demand by 25% in each is associated with a gradual increase in t s r max of 9.2% and only 5.6% using sulfur instead of thiosulfate. This can be of importance in a later development of processes since sulfur is highly toxic and volatile.

Influence of the Ci source on the production of methionine A main objective for the improvement of C. gl utami cum for methionine production is the Ci metabolism. Optimal production of methionine binds to the accumulation of identical molar amounts of glycine, which normally can not be reused (Figure 3). As shown, this can be overcome by the implementation of a glycine cleavage system (Figure 2 E). An alternative is given by the use of a carbon Ci source in addition to glucose. In this regard, formate was investigated involving different extensions of the metabolic network. This included the incorporation of an enzyme that catalyzes the formation of 10-formyl-THF from formate, ATP and THF as described for many organisms, for example bacilli (E.C. 6.3.4.3). Additionally, different steps for conversion of 10-formyl-THF into methyl-THF were implemented. All the reactions were agglomerated together in a global reaction that converts 10-formyl-THF in Methylene-THF bound to oxidation of 1 NADPH and 1 NADH. The use of formate plus glucose led to a slight increase in the theoretical maximum methionine yield of 3.3% compared to the situation with the sole use of glucose. Additionally, glycine was no longer accumulated when formate was supplied.

Influence of the combined use of different sources of sulfur and d on the production of methionine It was shown in the foregoing that the source of Ci and sulfur is important to maximize the maximum theoretical yield in carbon in the biotechnological production of methionine. Therefore it seems interesting to see if the benefits of Cx and sulfur sources can be combined. The studies involved the combination of thiosulfate and formate and the combination of sulfur and formate. For the combination of thiosulfate and formate, the maximum theoretical carbon yield was increased to 63.0% (Figure 3 D). This yield is still slightly lower than the theoretical maximum sulfur consumption yield (Figure 3 B). However, in contrast to sulfur, formate and thiosulfate are non-hazardous chemicals and probably join with reduced efforts concerning process safety. Combining the use of sulfur and formate resulted in? Met / s, max up to 69.6%. This is 6.2% higher than the maximum theoretical yield of the single sulfide consumption.

Influence of methanethiol and its dimethyl disulfide dimer as a combined source for sulfur and carbon C? on methionine production An interesting possibility of providing reduced sulfur and solving the problem of glycine accumulation is provided by providing methanethiol and its dimethyldisulfide dimer. It is known that C. gl utami cum can produce methanethiol under certain conditions (Bonnarme et al. (2000), Appl. Environ. Mi crobiol, 6612, 5514-7). It is assumed here that it is also capable of consuming methanethiol and its dimethyldisulfide dimer. It is also assumed that dimethyldisulfide dimer can be cleaved to methanethiol by the mentioned organisms such as but not limited to C. gl utamicum or E. coli A putative reaction was added to the network using direct methyl sulfhydrylation of O-acetyl-homoserine with methanethiol. This proposed new reaction eludes homocysteine and yields methionine directly. The use of methanethiol and its dimethyldisulfide dimer greatly increased the theoretical maximum methionine yield to 83.3% (Figure 3 F). This shows that this substrate is very useful potentially for the biotechnological production of methionine. The pathways involved in sulfur metabolism and MTHF formation may be dispensable for the production of methionine. Jan . coli, the theoretical maximum yield of methionine in methanethiol and its dimethyldisulfide dimer was 71.4% and thus was substantially lower compared to C. gl utamicum. The reason is the requirement of succinyl-CoA for production of methionine in this organism. This demands a high activity of the TCA cycle and the corresponding loss of carbon through C0. Compared to C. gl utami cum, the formation of C02 in E. Coli under these conditions is almost twice as high. Finally, the maximum theoretical yield can also be improved by integrating a transhydrogenase in the C. gl utamicum model with the metabolism of methanethiol. Under these conditions C. gl utamicum may be able to produce methionine from methanethiol and its dimethyl dimethyl sulfide and glucose with a maximum carbon yield of 85.5%. The above analysis in this manner has shown particularly that the use of glycine or alternative sources of the methyl group in methionine synthesis offer an important potential to optimize methionine production in C. gl utami cum. Additionally, it can be shown that the synthesis of methionine in E. coli is more dependent on an active transhydrogenase than C. gl utami cum. B) Genetic modification of C. glutamicum to increase the efficiency of methionine synthesis The goal of the following experiments is to apply the implications of previous theoretical findings to obtain a C. gl utami cum organism with increased efficiency of methionine synthesis Material and Methods Protocols for general methods can be found in Handbook on Corynebacterium glutamicum, (2005) eds .: L. Eggeling, M. Bott., Boca Raton, CRC Press, in Martin et al. (Biotechnology (1987) 5, 137-146), Guerrero et al.

(Gene (1994), 138, 35-41), Tsuchiya and Morinaga (Biotechnology (1988), 6, 428-430), Eikmanns et al. (Gene (1991), 102, 93-98), EP 0 472 869, US 4,601,893, Schwarzer and Pühler (Biotechnology (1991), 9, 84-87, Reinscheid et al. (Applied and Environmental Microbiology (1994), 60, 126-132), LaBarre et al (Journal of Bacteriology (1993), 175, 1001-1007) , WO 96/15246, Malumbres et al. (Gene (1993), 134, 15-24), in JP-A-10-229891, in Jensen and Hammer (Biotechnology and Bioengineering (1998), 58, 191-195), Makrides (Microbiological Reviews (1996), 60, 512-538) and in well-known textbooks of genetics and molecular biology.

Strains, Medians and Plasmids Strains can be taken for example from the following list: Corynebacterium glutamicum ATCC 13032, Corynebacterium acetoglutamicum ATCC 15806, Corynebacterium acetoacidophilum ATCC 13870, Corynebacterium thermoaminogenes FERM BP-1539, Corynebacterium melassecola ATCC 17965, Brevibacterium flavum ATCC 14067, Brevibacterium lactofermentum ATCC 13869, and Brevibacterium divaricatum ATCC 14020 or strains that have been derived from them such as Csrynejacteriu ?! glutamicum KFCC10065 DSM 17322 or Coryneba cteri um glutamicum ATCC21608 Recombinant DNA technology The protocols can be found in: Sambrook, J., Fritsch, E.F., and Maniatis, T., in Molecular Cloning: A Laboratory Manual, 3rd edition (2001) Cold Spring Harbor Laboratory Press, NY, Vol. 1, 2, 3, and Handbook on Corynebacterium glutamicum (2005) eds. L. Eggeling, M. Bott. , Boca Ratón, CRC Press.

Quantification of amino acids and methionine intermediates. The analysis is done by HPLC (Agilent 1100, Agilent, Waldbronn, Germany) with a guard cartridge and a Synergi 4μm column (MAX-RP 80 A, 150 * 4.6 mm) (Phenomenex, Aschaffenburg, Germany). Prior to injection the analytes are derived using o-phthaldialdehyde (OPA) and mercaptoethanol as a reducing agent (2-MCE). Additionally the sulfhydryl groups are blocked with iodoacetic acid. The separation is carried out at a flow rate of 1 ml / min using 40 mM NaH2P04 (eluent A, pH = 7.8, adjusted with NaOH) as polar phase and a methanol mixture water (100/1) as the non-polar (eluent B). The following gradient applies: Start 0% of B; 39 min 39% of B; 70 min 64% B; 100% B for 3.5 min; 2 min O% of B for equilibrium. The derivation at room temperature is automated as described in the following. Initially 0.5 μl of 0.5% of 2-MCE in bicine (0.5M, pH 8.5) are mixed with 0.5 μl of cell extract. Subsequently 1.5 μl of 50 mg / ml of iodoacetic acid in bicine (0.5M, pH 8.5) are added, followed by the addition of 2.5 μl of bicine buffer (0.5M, pH 8.5). The derivation is done by adding 0.5 μl of lOmg / ml of OPA reagent dissolved in 1/45/54 v / v / v of 2-MCE / MeOH / bicine (0.5M, pH 8.5). Finally, the mixture is diluted with 32 μl of H20. Between each of the pipetting steps in the above there is a waiting time of 1 min. A total volume of 37.5 μl is then injected into the column. Note that the analytical results can be significantly improved, if the needle of the automatic sampling device is periodically cleaned in the course of (for example, within the waiting time) and after sample preparation. Detection is performed by a fluorescence detector (340 nm excitation, 450 nm emission, Agilent, Waldbronn, Germany). For quantification, α-aminobutyric acid (ABA) was used as internal standard Definition of recombination protocol The following will describe how a strain of C. gl utamicum with increased efficiency of methionine production can be constructed by implementing the findings of the predictions in the above. Before the construction of the strain is described, a definition of an event / recombination protocol is given that will be used in the following. "Campbell within," as used herein, refers to a transformant of an original host cell in which a complete circular double-stranded DNA molecule (eg, a plasmid) has been integrated into a chromosome by a single event of homologous recombination (an inward crossing event), and which effectively results in the insertion of a linearized version of the circular DNA molecule into a first DNA sequence of the chromosome that is homologous to a first DNA sequence of the circular molecule of DNA. "Campbell-induced" refers to the linearized sequence of DNA that has been integrated into the chromosome of a "Campbell inside" transformant. A "Campbell inside" contains a duplication of the first homologous sequence of DNA, each copy of which includes and surrounds a copy of the crossing point of the homologous recombination. The name comes from Professor Alan Campbell, who first proposed this kind of recombination. "Campbell out," as used herein, refers to a cell descending from a "Campbell inside" transformant, in which a second homologous recombination event (an outward crossing event) has occurred between a second sequence. of DNA that is contained in the linearized inserted DNA of the "induced Campbell within" DNA, and a second DNA sequence of chromosomal origin, which is homologous to the second DNA sequence of the linearized insert, the second recombination event that results in the removal (shedding) of a portion of the integrated DNA sequence, but, importantly, that it also results in a portion (this can be as little as a single base) of the integrated DNA induced to the Campbell effect within that remains in the chromosome, so that compared to the original host cell, the "Campbell off" cell contains one or more intentional changes in the chromosome (for example, a single base substitution, its multi-base substitutions, insertion of a gene or heterologous DNA sequence, insertion of a copy or additional copies of a homologous gene or a modified homologous gene, or insertion of a DNA sequence comprising more than one of these aforementioned examples listed in the above). A "Campbell off" cell or strain is usually, but not necessarily, obtained by a counter-selection against a gene that is contained in a portion (the portion to be deslastered) of the DNA sequence "induced to Campbell effect within". ", for example the gene sa cB of Ba cillus subtilis, which is lethal when expressed in a cell that is grown in the presence of about 5% to 10% sucrose. Either with or without a counter-selection, a desired "Campbell out" cell can be obtained or identified by selecting the desired cell, using any selectable phenotype, such as, but not limited to, colony morphology, colony color, presence or absence of antibiotic resistance, presence or absence of a given DNA sequence by polymerase chain reaction, presence or absence of an auxotrophy, presence or absence of an enzyme, hybridization of nucleic acids in the colony, selection by antibodies , etc. The terms "Campbell inside" and "Campbell outside" can also be used as verbs in various sentences to refer to the method or process described in the above. It is understood that homologous recombination events that lead to a "Campbell inside" or "Campbell outside" can occur over a range of DNA bases within the homologous DNA sequence, and since the homologous sequences will be identical to each other for at least part of this interval, it is usually not possible to specify exactly where the crossing event occurred. In other words, it is not possible to specify precisely which sequence was originally from the inserted DNA, and which was originally chromosomal DNA. Moreover, the first homologous DNA sequence and the second homologous DNA sequence are usually separated by a region of non-partial homology, and it is this region of non-homology that remains deposited on a chromosome of the "Campbell off" cell. For practical reasons, in C gl utamicum, the first and second typical homologous DNA sequence is at least about 200 base pairs in length, and can be up to several thousand base pairs in length, however, the procedure can be done with shorter or longer sequences. For example, a length for the first and second homologous sequences may vary from about 500 to 2000 bases, and obtaining a "Campbell out" from a "Campbell within" is facilitated by arranging the first and second homologous sequences for which are of approximately the same length, preferably with a difference of less than 200 base pairs and more preferably with the shorter of the two being at least 70% longer than the longest in base pairs.

Construction of the methionine producing strain Example 1: Generation of the methionine-producing start strain M2014 strain C. gl utamicum strain ATCC 13032 was transformed with DNA A (also referred to as pH273) (SEQ ID NO: 1) e " induced Campbell effect inside "to yield a strain" Campbell inside ". Figure 6 shows a scheme of the plasmid pH273. The "Campbell in" strain was then "induced to Campbell effect out" to yield a "Campbell off" strain, M440, which contains a gene that encodes a homoserin dehydrogenase-resistant enzyme (homfbr). The resulting homoserine dehydrogenase protein included an amino acid change where S393 changed to F393 (referred to as Hsdh S393F). Strain M440 was subsequently transformed with DNA B (also referred to as pH373) (SEQ ID NO: 2) to yield a "Campbell in" strain. Figure 6 depicts a scheme of plasmid pH373. The "Campbell in" strain was then "induced to Campbell effect out" to yield a "Campbell off" strain, M603, which contains a gene encoding a feedback-resistant aspartate kinase enzyme (Askfbr) (encoded by lysC). In the resulting aspartate kinase protein, T311 changed to 1311 (referred to as LysC T311I). It was found that strain M603 produced about 17.4 mM of lysine, while strain ATCC13032 did not produce a measurable quantity of lysine. Additionally, strain M603 produced about 0.5 mM homoserin, compared to a non-measurable amount produced by strain ATCC13032, as summarized in Table 3. Table 3: Amounts of homoserin, O-acetylhomoserin, methionine and lysine produced by strains ATCC 13032 and M603 Strain M603 was transformed with DNA C (also referred to as pH304, a scheme of which is depicted in Figure 6) (SEQ ID NO: 3) to yield a "Campbell inside" strain, which was then "induced to Campbell effect". out "to yield a strain" Campbell out ", M690. Strain M690 contained a PgroES promoter upstream of the metH gene (referred to as P497 metH). The P497 promoter sequence is depicted in SEQ ID NO: 11. The M690 strain produced about 77.2 mM lysine and about 41.6 mM homoserine, as shown in the following in Table 4. Table 4: Amounts of homoserine , O-acetyl homoserine, methionine and lysine produced by strains M603 and M690 Strain M690 was subjected to mutagenesis subsequently as follows: an overnight culture of M690, grown in BHI medium (BECTON DICKINSON), was washed in 50mM citrate buffer pH 5.5, treated for 20 min at 30 ° C with N -methyl-N-nitrosoguanidine (10 mg / ml in 50mM citrate pH 5.5). After that treatment, the cells were washed again in 50 mM citrate buffer pH 5.5 and plated in a medium containing the following ingredients: (all the amounts mentioned are calculated for 500 ml of medium) lOg of (NH4) ) 2S04; 0.5g of KH2P04; 0.5g of K2HP04; 0.125g of MgSO4 * 7H20; 21g of MOPS; 50 mg of CaCl2; 15 mg of protocatechuic acid; 0.5 mg of biotin; 1 mg of thiamine; and 5 g / 1 of D, L-ethionine (SIGMA CHEMICALS, CATALOG # E5139), adjusted to pH 7.0 with KOH. In addition, the medium contained 0.5 ml of a trace metal solution composed of: 10 g / 1 of FeS04v7H20; 1 g / 1 of MnS04 * H20; 0.1 g / 1 of ZnS04 * 7H20; 0.02 g / 1 CuS04; and 0.002 g / 1 of NiCl2 * 6H20, all dissolved in 0.1 M HCl. The final medium was sterilized by filtration and 40 milliliters of 50% sterile glucose solution (40 ml) and sterile agar at a final concentration of 1.5% were added to the medium. The final medium containing agar was poured onto agar plates and labeled as the minimal-ethionine medium. Strains subjected to mutagenesis were dispersed on the plates (minimal-ethionine) and incubated for 3-7 days at 30 ° C. The clones that grew on the medium were isolated and recultivated in pure isolation on the same minimal-ethionine medium. Several clones were selected for methionine production analysis. The methionine production was analyzed as follows. The strains were grown on CM-agar medium for two days at 30 ° C, which contained: 10 g / 1 of D-glucose, 2.5 g / 1 of NaCl; 2 g / 1 urea; 10 g / 1 Bacto Peptone (DIFCO); 5 g / 1 of Yeast Extract (DIFCO); 5 g / 1 of Beef Extract (DIFCO); 22 g / 1 Agar (DIFCO); and which was autoclaved for 20 min at about 121 ° C. After the strains were grown, the cells were removed by scraping and resuspended in 0.15 M NaCl. For the main culture, a suspension of cells removed by scraping was added to a starting OD of 600 nm to about 1.5 to 10 ml of Medium II (see below) together with 0.5 g of solid CaC03 and sterilized by autoclaving ( RIEDEL DE HAEN) and the cells were incubated in a 100 ml shake flask without deflectors for 72 h on an orbital shaking platform at about 200 rpm at 30 ° C. Medium II contained: 40 g / 1 of sucrose; 60 g / 1 of total molasses sugar (calculated for the sugar content); 10 g / 1 of (NH4) 2S04; 0.4 g / 1 MgSO4 * 7H20; 0.6 g / 1 of KH2P04; 0.3 mg / l thiamine * HCl; 1 mg / l of biotin; 2 mg / l FeS04; and 2 mg / l of MnS04. The medium was adjusted to pH 7.8 with NH 4 OH and sterilized by autoclaving at about 121 ° C for about 20 min.). After autoclaving and cooling, vitamin Bi2 (cyanocobalamin) (SIGMA CHEMICALS) was added from a sterile stock solution by filtration (200 μg / ml) to a final concentration of 100 μg / l. Samples were taken from the medium and analyzed for amino acid content. The amino acids produced, including methionine, were determined using the Agilent amino acid method in an Agilent 1100 Series LC System HPLC. (AGILENT). A pre-column derivation of the sample with ortho-phthalaldehyde allowed the quantification of amino acids produced after separation on a Hypersil AA (AGILENT) column. Clones that showed a methionine titre that was at least twice that in M690 were isolated. One clone, used in additional experiments, was named M1197 and deposited on May 18, 2005, in the DSMZ strain collection as strain number DSM 17322. The production of amino acids by this strain was compared with that by strain M690, as it is summarized in the following in Table 5.

Table 5: Amounts of homoserin, O-acetylhomoserin, methionine and lysine produced by strains M690 and M1197 Strain M1197 was transformed with DNA F (also referred to as pH399, a scheme of which is depicted in Figure 7) (SEQ ID NO. : 4) to yield a "Campbell inside" strain, which subsequently "induced Campbell effect out" to yield the M1494 strain. This strain contains a mutation in the gene for homoserine kinase, which results in an amino acid change in the enzyme homoserine kinase resulting from T190 to A190 (referred to as HskT190A). The production of amino acids by strain M1494 was compared to the production by strain M1197, as summarized in the following in Table 6.

Table 6: Amounts of homoserin, O-acetylhomoserin, methionine and lysine produced by strains M1197 and M1494 Strain M1494 was transformed with DNA D (also referred to as pH484, a scheme of which is shown in Figure 7) (SEQ ID NO: 5) to yield a "Campbell within" strain, which subsequently was "induced to Campbell effect". outside "to yield the M1990 strain. Strain M1990 overexpresses a met Y allusing a groES promoter and an EFTU promoter (Tu elongation factor) (referred to as P497 P1284 met Y). The sequence of P4g7 P? 2s is set forth in SEQ ID NO: 13. The production of amino acids by strain M1494 was compared to the production by strain M1990, as summarized in the following in Table 7.

Table 7: Amounts of homoserin, O-acetylhomoserin, methionine and lysine produced by strains M1494 and M1990 Strain M1990 was transformed with DNA E (also referred to as pH 491, a scheme of which is depicted in Figure 7) (SEQ ID NO: 6) to yield a "Campbell within" strain, which was then "induced to Campbell effect". out "to yield a strain" Campbell out "M2014. Strain M2014 overexpresses a metA allusing a superoxide dismutase promoter (referred to as P3119 metA). The sequence of P3119 is set forth in SEQ ID NO: 12. The production of amino acids by strain M2014 was compared to the production by strain M2014, as summarized in the following in Table 8.

Table 8: Amounts of homoserin, 0-acetylhomoserin, methionine and lysine produced by strains M1990 and M2014 Example 2: Experiments in agitation and HPLC assay Experiments in shake flasks, with the standard Molasses Medium, were performed with strains in duplicate or quadruplicate. The Molasses Medium contained in a liter of medium: 40 g of glucose; 60 g of molasses; 20 g of (NH) 2S04; 0.4 g of MgSO4 * 7H20; 0.6 g of KH2P04; 10 g of yeast extract (DIFCO); 5 ml of 400 mM threonine; 2 mg FeS04.7H20; 2 mg of MnSO4H20; and 50 g of CaCO3 (Riedel-de Haen), with the volume compensated with ddH20. The pH was adjusted to 7.8 with 20% NH4OH, 20 ml of agitated medium continuously (in order to keep the suspended CaC03) were added to 250 ml warp shake flasks with baffles and the flasks were sterilized by autoclaving for 20 min. . Subsequent to the autoclave sterilization, 4 ml of "4B solution" were added per liter of the base medium (or 80 μl / flask). The "solution 4B" contained per liter: 0.25 g of thiamine hydrochloride (vitamin Bl), 50 mg of cyanocobalamin (vitamin B12), 25 mg of biotin, 1.25 g of pyridoxine hydrochloride (vitamin B6) and was subjected to buffer state with 12.5 mM of KP04, pH 7.0 to dissolve the biotin, and sterilized by filtration. The cultures were grown in flasks with baffles covered with Bioshield paper secured by rubber bands for 48 hours at 28 ° C or 30 ° C and at 200 or 300 rpm in a New Brunswick Scientific floor shaker. Samples were taken at 24 hours and / or 48 hours. The cells were removed by centrifugation followed by dilution of the supernatant with an equal volume of 60% acetonitrile and then by membrane filtration of the solution using Centricon Centrifuge columns of 0.45 μm. The filtrates were analyzed using HPLC for methionine concentrations, glycine plus homoserin, 0-acetylhomoserin, threonine, isoleucine, lysine, and other indicated amino acids. For the HPLC assay, the filtered supernatants were diluted 1: 100 with 1 mM Na2EDTA filtered at 0.45 μm and 1 μl of the solution was derived with OPA reagent (AGILENT) in Borate buffer (80 mM NaB03, 2.5 mM EDTA, pH 10.2 ) and injected into a 200 x 4.1 mm Hypersil 5μ AA-ODS column introduced in an Agilent 1100 HPLC series equipped with a G1321A fluorescence detector (AGILENT). The excitation wavelength was 338 nm and the emission wavelength monitored was 425 nm. Standard amino acid solutions were subjected to chromatography and used to determine retention times and standard peak areas for the various amino acids. Chem Station, the accompanying software package provided by Agilent, was used for instrument control, data acquisition and data manipulation. The hardware was an HP Pentium 4 computer that supports Microsoft Windows NT 4.0 updated with a Microsoft Service Pack (SP6a).

Example 3: Generation of a microorganism containing a sulfa to reduction pathway destabilized in its regulation Plasmid pOM423 (SEQ ID NO: 7) was used to generate strains containing a destabilized sulfate reduction pathway in its regulation. Specifically, a construct of divergent promoters PL and PR of lambda phage from E. coli was used to replace the native divergent promoters of the sulphate reduction regution. Strain M2014 was transformed with pOM423 and selected for kanamycin resistance (Campbell within). After a contra-selection with sacB, kanamycin-sensitive derivatives were isolated from the transformants (Campbell outside). These were subsequently analyzed by PCR to determine the structures of the promoters of the sulfate reduction regu lation. The isolates containing the divergent PL-PR promoters were named OM429. Four isolates of OM429 were analyzed for sulphate reduction using the DTNB strip test and for methionine production in shake flask assays. To estimate the relative production of sulfur using the DTNB strip test, a strip of filter paper was soaked in an Ellman reagent solution (DTNB) and suspended over a shake flask culture of the strain to be tested for 48 hours. The hydrogen sulfide produced by the growing crop reduces the DTNB, producing a yellow color that roughly proportional to the amount of H2S generated. In this way, the intensity of the color produced can be used to obtain an approximate estimate of the relative sulphate reduction activity of various strains. The results (Table 10) show that two of the four isolates showed relatively high levels of sulfate reduction. These same two isolates also produced the highest levels of methionine. The cultures were grown for 48 hours in standard molasses.

Table 10. Methionine production and sulphate reduction by isolations of OM429 in shake flask cultures Strain Met promoters Sulphate regution test (g / 1) DTNB M2014 Native 1.1 - OM429-1 PL / P 1.1 - -2 1.1 - -3 1.3 ++ -4 1.4 ++ Experiment 4: Strains that have the glycine cleavage operon (gcv) of E. coli. The production of methylene tetrahydrofolate, from serine by the enzyme GlyA (R38) which is necessary for methionine biosynthesis from glucose, also renders glycine as a by-product. In overproducing methionine strains, the amount of glycine produced will be greater than the requirement for protein synthesis. In this way, according to the model in the above, the inclusion of the GCS in C. gl utami cum should result in enhanced efficiency of methionine synthesis. Jan . coli and B. Subtilis, if glycine is present higher than that required for protein synthesis, is cleaved to give a second equivalent of methylenetetrahydrofolate by the glycine cleavage enzyme system. Jan . coli, the glycine cleavage system involves four different proteins. Three of these are encoded by the gcvTHP operon. The fourth subunit is lipoamide dehydrogenase, which is borrowed from multiple subunit pyruvate dehydrogenase. C. gl utami cum does not seem to have a glycine cleavage system. No homologs of the E. coli Gcv proteins were found in the genome of C. glutamicum, although C. gl utamicum has the usual multiple subunit pyruvate dehydrogenase. As a result, the production of methionine in C. gl utamicum results in concomitant production of glycine, which appears in culture supernatants. In this way an attempt was made to implement a GCS in C. glutamicum and to recycle glycine in methylenetetrahydrofolate, as is done in E. coli and B. subtilis. As a first step toward this goal, the gcvTHP operon from E. coli was amplified by PCR without its native promoter, and cloned downstream of the P497 promoter in pOM218, which is a vector of low copy number E. coli designed to integrate expression cassettes in bioB in C. gl utami cum. It was assumed that the fourth required subunit of pyruvate dehydrogenase can be delivered from the host organism which is C. gl utamicum. The resulting plasmid, pOM229 (Figure 8, SEQ ID NO: 8), was transformed into the initiator organism, strain M2014 and successfully induced to Campbell effect outside to give the strains named OM212. These strains were then cultured. The following medium was used: 40 g / 1 glucose, 60 g / 1 of molasses with a sugar content of 45%, 10 g / 1 of (NH4) 2S04, 0.4 g / 1 of MgSO4 * 7H20, 2 mg / l of FeS04, 2 mg / l of MnS04, 1.0 mg / l of thiamine, 1 mg / l of biotin. The pH was adjusted to pH 7.8 with 30% NH0H, and the medium was sterilized by autoclaving for 20 minutes. After autoclaving: 200 Mg / l of B12, 2 mM of L-threonine, 2 ml of 0.5 g / ml of CaCO3 per 20 ml of medium. Phosphate buffer pH 7.2 SE was added to 200 mM of a 2 M stock. In cultures in shake flasks, an isolation, OM212-1 was analyzed as explained above. The results showing an increase in methionine production and a decrease in glycine plus homoserine are shown in Table 11.

Table 11 Production of methionine by M2014 derivatives containing P4g7 gcvTHP (E. coli) integrated in bioB, in shake flask cultures grown in molasses plus CaC03 medium.

[Gly + [O-Ac- [Lys] [Met] Strain New characteristic Hse] Hse] g / i g / l g / l g / l M2014 progenitor 0.66 1.4 3.3 0.64 0.75 1.6 3.4 EYE Pr 0.67 OM212-1 pOM229 0.67 1.7 3.8 0.74 Pw gcvTHP 0.58 1.8 3.7 EYE @ bioB Pr 0.72 It was observed that the carbon yield of strain M2014 was 0.0103 Mol of methionine / mol of sugar while the strain OM212-1 had a carbon yield of 0.011 Mol of methionine / mol of sugar. In another embodiment the subunit of the glycine cleavage system not encoded by the gcvTHP operon, ie the lpdA gene (SEQ ID NO: 10), which codes for lipoamide dehydrogenase is cloned from host E. coli The gene is amplified without its natural promoter and the P 97 promoter is added in its place. The resulting fragment is cloned into the shuttle vector pOM229 of E. coli-C glutamicum in addition to the gcvTHP operon.

Experiment 5: An in vivo assay for a functional glycine cleavage system.

The serA gene of C. glutami cum was generated by PCR and cloned into pC INT open with Swa I to give the plasmid pOM238. Then, a fragment with blunt ends containing a gram-positive spectinomycin resistance gene (spc) expressed from C. glutamicum P497 was ligated into open pOM238 with Ale I. An isolation containing the spc gene in the same orientation to be named pOM253 (see Figure 9, SEQ ID NO: 9). pOM253 can be used to create an interruption-deletion in the serA gene of any strain of C. gl utamicum. pOM253 was transformed into the M2014 strain of C. gl utamicum, selecting for kanamycin resistance, to give the OM264K strain "induced to Campbell effect inside". OM264K was "induced to Campbell effect out" when selecting for sucrose resistance (BHI + 5% sucrose) and spectinomycin resistance (BHI + 100 mg / l spectinomycin) to give strain OM264, which is an auxotroph of serine , threonine, and biotin. Strain OM264 can be transformed with the plasmid pOM229, or another plasmid (or plasmids) that supplies the glycine cleavage pathway (Gcv). If the glycine cleavage pathway is active, then the resulting serAX Gcv + strain will be able to grow in minimal medium containing glycine, threonine, and biotin, since methylenetetrahydrofolate will be generated by the glycine cleavage system, and the gene product glyA, serine hydroxymethyl transferase (SHMT), will be able to produce serine by running the SHMT reaction in the reverse direction, using glycine and methylenetetrahydrofolate as substrates. If necessary, a gene encoding lipoamide dehydrogenase, for example, the lpd gene (also called IpdA; Sec No: 10) of E. coli can be cloned and transformed into the strain described above to provide the fourth subunit necessary for the glycine cleavage system. The genes encoding the glycine cleavage systems of organisms other than E. coli can also be cloned by PCR or complementation as described above and used to deliver a functional glycine cleavage system in C. glutamicum. For example, the genes of Bacill us subtilis, gcvH, gcvPA, gcvPB, gcvT and pdhD, which encode a glycine cleavage system of five subunits (glycine decarboxylase is comprised of two subunits in B. subtilis, encoded by gcvPA and gcvPB, while in E. coli these two functions are combine in a subunit encoded by gcvP), or any other suitable set of genes can be used. The only requirement is that the system works in C. gl utami cum at a sufficient level to convert excess glycine (produced as a result of methionine biosynthesis) to methylenetetrahydrofolate.

Experiment 6: deactivation of pyruvate kinase in C. glutamicum Elemental analysis indicated that a down regulation of pyruvate kinase (R19) can lead to an increased efficiency of methionine synthesis (see for example Figure 3). To investigate the effect of the inactivation of pyruvate kinase, a strain of C. gl utami cum that produces lysine was analyzed. If in fact an increase in lysine production were observed, this should also be indicative of an increased synthesis of methionine, since the formation of lysine is preceded by the formation of aspartate, aspartate-phosphate, etc. An increase in lysine production must therefore be preceded by an increase for example in aspartate. Since aspartate is also one of the precursors of methionine production, an increased amount of aspartate must also lead to increased methionine synthesis. A comparison of strains between C. gl utamicum lysCfbr and C gl utamicum lysCfbr? Pyk was carried out. C. gl utami cum lysCfbr is a mutant that carries a point mutation in the gene encoding aspartokinase (Kalinowski et al (1991), Mol.Microbiol.5 (5), 1197-1204). This strain was then used to remove pyruvate kinase (C. gl utami cum lysCfbr? Pyi). Both strains were grown in shaker flasks in minimal media and the carbon yields were determined for biomass, lysine and by-products. Based on the mean value of two independent experiments, it was observed that the lysine yields for the inactivation of pyruvate kinase were increased by 7.5-12.1%. This corresponds to an increase of approximately 62%. In conclusion, an inactivation of pyruvate kinase leads to an increased synthesis of lysine and correspondingly must also lead to increased methionine synthesis. However, using inactivation of pyruvate kinase to produce methionine may not have been expected to increase the synthesis of methionine, since methionine itself uses an active pyruvate kinase if you take into account the common knowledge about metabolic networks.

Experiment 7 Comparison of uptake in the use of different sulfur sources Elemental analysis had shown that the efficiency of methionine synthesis surprisingly was dependent on the state of reduction of the sulfur source. As explained above, for each NADPH saved an increase in methionine synthesis efficiency of 4.6% can be expected. However, until now there are only preliminary and incomplete data regarding the growth and use of different sulfur sources by C. glutamicum.

In order to test whether the culture of C. gl utamicum in different carbon sources actually leads to an increased level of methionine synthesis efficiency, the following experiments were performed. A wild type strain of C. gl utamicum and the mutant? JncbR were cultured in sulfate and thiosulfate in shake flasks. For that purpose, the corresponding sulfur sources were added in molarity concentrations identical to a minimum medium CG12 ^ without sulfur. The CG12 ^ medium comprises per liter: 20 g of glucose, 16 g of K2HP04, 4 g of KH2P04, 20 g of (NH4) 2S04, 300 mg of 3,4-dihydroxybenzoic acid, 10 mg of CaCl2, 250 mg of MgSO4 7 H2O, 10 mg of FeS04 * 7 H20, 10 mg of MnS04 * H20, 2 mg of ZnS04 * 7 H20, 200 μg of CuS04 * 5 H20, 20 μg of NiCl2 * 6 H20, 20 μg of Na2Mo0 * 2 H20, 100 μg of cyanocobalamin (Vitamin B? 2), 300 μg of thiamin (vitamin Bi), 4 μg of pyridoxal phosphate (vitamin Be) and 100 μg of biotin (vitamin B7). In the case of the sulfur free GC12 ^ medium, all the sulphates were replaced by chlorines used in concentrations so that the concentrations of the corresponding cations could not change. The following salts were used: MgCl2 * 6 H20 (SO42"<0.002%, Sigma); ZnCl2 (SO42 ~ <0.002%, Sigma); NH4C1 (SO42 ~ <0.002%, Fluka); MnCl4 * 4H20 ( SO42"< 0.002%, Sigma) and FeCl2 * 4 H20 (SO42" < 0.01%, Sigma).

The culture of C. gl utami cum was carried out in agitator flasks with indentations at 30 ° C and 250 upm in agitator cabinets (Multitron, Infors AG, Bottmingen, Switzerland) . In order to avoid an oxygen limitation, the flasks were filled to a maximum of 10% with medium. It is known that cysteine synthase CysK (R45 and R45a) and cystathionine -? - synthase MetB (R46) are overexpressed in C. gl u tami cum? McbR (Rey et al. (2003) vide supra). It was found that both strains can grow in sulfate and thiosulfate. The highest growth rate was observed for the wild type with μmax = 0.44h_1 in sulfate. Sulfate in this manner seems to be the preferred source of sulfur for C. gl utamicum. Thiosulfate was also used for C. glutamicum, at the observed lower growth rate of μma? = 0.31 h "1. However, an increase in biomass was observed for the wild type from 0.35 gg-1 to 0.60 gg" 1 if the sulfate was replaced by thiosulfate. In the case of deactivation? IOabR, the yield in biomass was increased even from 0.42 gg "1 to 0.51 gg" 1 if the sulfate was replaced by thiosulfate. This corresponds to an increase in performance of 13% and 21%. In this way, replacing sulfate with thiosulfate actually leads to a reduction in ATP and NADPH which in turn has a positive effect on carbon yield.

Since a reduced amount of sugar / glucose is needed for the production of biomass, more sugar / glucose is available for the production of methionine. Thus, a change from sulfate to thiosulfate should in fact lead to increased yields of methionine synthesis and this effect should be even more pronounced if the use of thiosulfate as the sulfur source is combined with an increase in metabolic flux through the metabolic pathways preferred by genetic manipulation.

Legends of Figures: Figure 1: Network of stoichiometric reactions of the wild type of C. glutamicum applied for elemental mode analysis. An arrow in two directions represents reversible reactions. The external metabolites are presented in gray boxes. Figure 2: Analysis of metabolic pathways of C. gl utamicum and E. coli for methionine production: yield in carbon for biomass and methionine for the elementary modes obtained from C. gl u tamicum wild type (A), E. coli wild type (B), C. gl utami cum mutant with active transhydrogenase (C), E. mutant coli lacking transhydrogenase (D), mutant C. glutami cum with glycine cleavage active system (E), mutant E. coli lacking glycine cleavage system (F). The given number indicates the maximum theoretical carbon yield for methionine for each scenario. The narrow line connects the modes with maximum yields in biomass and maximums of methionine. Figure 3: Flow distribution of the wild type of C. gl utami cum with maximum theoretical yield of methionine-carbon. All flows are given as molar fluxes relative to glucose uptake. Figure 4: Wild type flow distribution of E. col i with maximum theoretical yield of methionine-carbon. All flows are given as molar fluxes relative to glucose uptake. Figure 5: Analysis of metabolic pathways of C. glutamicum for methionine production with different carbon and sulfur sources: carbon yield for biomass and methionine for the elemental modes obtained from C glutami cum using thiosulfate (A), thiosulfate and formate (B ), sulfur (C), sulfide and formate (D), formate (E) and methanethiol or its dimethyl dimethyl sulfide (F). The given number indicates the maximum theoretical carbon yield for methionine for each scenario. The straight line connects the modes with maximum biomass yields and methionine maxima. Figure 6 shows the vector pH 273, pH 373 and pH 304 Figure 7 shows the vector pH 399, pH 484 and pH 491 Figure 8 shows the vector pOM 229. Figure 9 shows the vector pOM 253. Figure 10 shows a preferred embodiment of optimized metabolic flux with respect to methionine synthesis.

Abbreviations: G6P = Glucose-6-phosphate F6P = Fructose-6-phosphate F-16-BP - Fructose-1, 6-bisphosphate ASP = Aspartic acid ASP-P = Aspartyl-phosphate ASP-SA = Aspartate-semialdehyde HOM = Homoserine O-AC-HOM = O-acetyl-homoserine HOMOCYS = homocysteine 3-PHP = 3-Phosphonoxypyruvate SER-P = 3-Fosfoserine SER = Serine O-AC-SER = O-acetyl-serine CYS = Cysteine CYSTA = Cystathionine GA3P = Glyceraldehyde 3-phosphate DAHP = Dihydroxyacetone phosphate 13-PG = 1,3-Bisphospho-glycerate 3-PG = 3-Phospho-glycerate 2-PG = 2-Phospho-glycerate AC-CoA = Acetyl coenzyme A PYR = Pyruvate PEP = Phosphoenol -piruvate CIT = Citric acid OAA = Oxaloacetate Cis-ACO = cis-Aconite ICI = Iso-citric acid 2-OXO = 2-Oxoglutarate GLU = Glutamate SUCC-CoA = Succinyl coenzyme A SUCC = Succinate FUM = Fumarate MAL = Malate GLYOXY = Glioxylate H2S03 = Sulfite H2S = Hydrogen sulfide 6-P-Gluconate = 6-Phospho-gluconate GLC-LAC = 6-Phospho-glucono-l, 5-lactone RIB-5P = Ribulose 5-phosphate RIB0-5P = Ribose 5-phosphate XYL-5P = Xylulose 5-phosphate S7P = Sedoheptulose 7-phosphate E-4P = Erythrose 4-phosphate MET = L-Methionine NADP = Nicotinamide dinucleotide phosphate -oxidated adenine NADPH = reduced Nicotinamide-adenine dinucleotide phosphate ACETAT = H-CoA acetate = Coenzyme A FAD = oxidized Flavin-adenine dinucleotide FADH = Flavin-adenine reduced dinucleotide ATP = 5 'Adenosine -triphosphate ADP = Adenosine 5' -NAD phosphate = Nicotinamide-oxidized adenine dinucleotide NADH = Nicotinamide-adenine reduced dinucleotide M-THF = 5-Methyltetrahydrofolate THF = Tetrahydrofolate GDP = Guanosine 5'-GTP phosphate = Guanosine 5'-GLC triphosphate = METex glucose = Excreted methionine 02 = NH3 oxygen = Ammonia C02 = Carbon dioxide S04 = Sulphate GLYCINE = Glycine HPL = protein-H-lipoylisine Methyl-HPL = protein-HS-aminomethyl-dihydrolipoylisine Reactions The following reactions are carried out by the enzymes Rl to R80: Rl: PEP + GLC = PYR + G6P. R2: G6P = F6P. R3: G6P + NADP = GLC-LAC + NADPH. R4: GLC-LAC = 6-P-Gluconate. R5: 6-P-Gluconate + NADP = RIB-5P + C02 + NADPH. R6: RIB-5P = XYL-5P. R7: RIB-5P = RIBO-5P. R8: S7P + GA3P = RIBO-5P + XYL-5P. R9: S7P + GA3P = E-4P + F6P. RIO: F6P + GA3P = E-4P + XYL-5P. Rll: ATP + F6P = ADP + F-16-BP. R12: F-16-BP = F6P. R13: F-16-BP = GA3P + DAHP. R14: DAHP = GA3P. R15: GA3P + NAD = 13-PG + NADH. R16: ADP + 13-PG = ATP + 3-PG. R17: 3-PG = 2-PG.

R18: 2-PG = PEP. R19: PEP + ADP = PYR + ATP. R20: PYR + H-CoA + NAD = AC-CoA + NADH + C02.

R21: AC-CoA + OAA = CIT + H-CoA. R22: CIT = Cis-ACO. R23: Cis-ACO = ICI. R24: ICI + NADP = 2-OXO + C02 + NADPH. R25: 2-0X0 + NH3 + NADPH = GLU + NADP. R26: 2-0X0 + NAD + H-CoA = SUCC-CoA + NADH + C02 R27: SUCC-CoA + GDP = SUCC + H-CoA + GTP. R28: SUCC + FAD = FUM + FADH. R29: FUM = BAD. R30: MAL + NAD = OAA + NADH. R31: ICI = GLYOXY + SUCC. R32: GLYOXY + AC-CoA = MAL + H-CoA. R33: PYR + ATP + C02 - OAA + ADP. R34: PEP + C02 = OAA. R35: OAA + ATP = PEP + ADP + C02. R36: OAA + ADP = PYR + C02 + ATP. R37: OAA + GLU + NADPH = ASP + 2-0X0 + NADP.

R38: THF + SER = MTHF + GLYCINE. R39: ASP-SA + NADPH = HOM + NADP. R40: HOM + SUCC-CoA = 0-SUCC-HOM + H-CoA. R41: 3-PG + NAD = 3-PHP + NADH. R42: 3-PHP + GLU = SER-P + 2-0X0.

R43: SER-P = BE. R44: SER + AC-CoA = O-AC-SER + H-CoA. R45: O-AC-SER + H2S = CYS + ACETAT. R45a: H2S203 + O-Ac-SER = S-Sulfocysteine + ACETAT R46: CYS + O-SUCC-HOM = CYSTA + SUCC. R47: ASP + ATP = ASP-P + ADP. R48: ASP-P + NADPH = ASP-SA + NADP. R49: O-Acetyl-homoserine + H2S = Homocysteine + acetic acid R50: ATP + ACETAT = ADP + acetyl-phosphate. R51: acetyl-phosphate + H-CoA = AC-CoA. R52: HOMOCYS + MTHF = MET + THF. R53: MET = METex. R54: CYSTA = HOMOCYS + NH3 + PYR. R55: S04 + 2 ATP + NADPH = H2S03 + 2 ADP + NADP. R56: ATP = ADP. R57: MAL + NADP = PYR + C02 + NADPH. R58: H2S03 + 3 NADPH = H2S + 3 NADP. R59: 2 NADH + 02 + 4 ADP = 2 NAD + 4 ATP. R60: 2 FADH + 02 + 2 ADP = 2 FAD + 2 ATP. R61: 6965 NH3 + 233 S04 + 206 G6P + 72 F6P + 627 RIBO -5P + 361 E-4P + 129 GA3P + 1338 3-PG + 720 PEP + 2861 PYR + 2930 AC -CoA + 1481 OAA + 1078 2-OXO + 16548 NADPH = BIOMASS + 16543 NADP + 2930 H-CoA + 1678 C02. R62: ADP + GTP = ATP + GDP.

R70: NADPH + NAD = NADP + NADH. R71: GLYCINE + HPL = Methyl-HPL + C02. R72: Methyl-HPL + THF = HPL + MTHF + NH. R73: 1 Thiosulfate (S2032") + 1 NAD (P) H = 1 Sulfite + 1 Sulfide + 1 NAD (P) R74: Sulfite + 3 NAD (P) H = Sulfide + 3 NAD (P) R75: ATP + Formate + THF = ADP + Orthophosphate + 10-formyl-THF R76: 5, 10-Methenyl-THF + NADPH = 5, 10-Methylene-THF + NADP R77: O-Acetyl-homoserine + methanethiol = methionine + acetate R78: 5, 10-Methylene-THF + NADP (H) = Methyl-THF R79: formyl-tetrahydrofolate = formate + tetrahydrofolate R80: sulfate + 1 NAD (P) H + 1 ATP + 1 G (A) TP = sulfite + 1 NAD (P ), IPPI, 1 G (A) DP + adenylate + P R81: 3 NADH + 3 NADP + + ATP = 3 NAD + + 3 NADPH R82: H2S203external + ATP = H2S203intemo + ADP The model of C. gl utamicum wild type (compare figure 1) - Reactions and Enzymes: Rl System phospho-transferase R2 G6P-isomerase R3 G6P-DH R4: Lactonase R5: Gluconate-DH R6: Ribose-5-P-epimerase R7 : Ribose-5-P-isomerase R8: Transketolase 1 R9: Transaldolase RIO: Transketolase 2 Rll: Phosphofruct kinase R12: Fructosebisphosphatase R13: Fructosebisphosphate-aldolase R14: Triosyphosphate isomerase R15: 3-phosphoglycerate-Kinase R16: PG-kinase R17: PG-mutase R18: PEP-hydrolase R19: PYR-kinase R20: PYR-DH R21: CIT-synthase R22: ACO-hydrolase R23: ACONITASE R24: Isocitrate-DH R25: Glutamate-DH R26: 2-OXO-DH R27: SUCC-CoA-synthase R28: SUCC-DH FUMARASE MAL-DH ICI-lyase MAL-synthase PYR-carboxylase PEP-carboxylase PEP-carboxykinase OAA-decarboxylase ASP-transaminase 10 Synthesis 1 of M-THF HOM -DH HOM-transacetylase PG-DH Phosphoserine transaminase Phosphoserine phosphatase Serine transacetylase Cysteine synthase Cystathionine synthase Aspartokinase 20 ASP-P-DH O-Ac-HOM sulfhydrylase ACETAT-kinase Phosphotransacetylase MET synthase (MetE / H) Methionine exporter R54 Cystathionine-D-lyase R55 ATP-sulfurylase R56 Hydrolysis of ATP R57 Magmatic enzyme R58 Sulfite-reductase R59 Respiratory chain 1 R60 Respiratory chain 2 R61 Biomass formation R62 GTP-ATP-Phosphotransferase Type of reaction (reversible or irreversible): Reversible. R2r R6r R7r R8r R9r R2R4 R5 R12 R12 R16 R20 R20 R24 R25 R26 R26 R27 R32 R32 R32 R33 R34 R36 R36 R38 R38 R39 R40 R43 R44 R45 R46 R47 R48 R48 R49 R50 R51 R52 R53 R54 R55 R56 R57 R58 R59 R60 R61 R62 Metabolites (internal or external): Internal: G6P F6P F-16-BP ASP ASP-P ASP-SA HOM ATP O-AC-HOM HOMOCYS 3-PHP SER-P BE O-AC-SER CYS CYSTA GA3P DAHP 13-PG 3-PG 2-PG AC-CoA PYR PEP CIT OAA CIS-ACO ICI 2-OXO GLU SUCC-CoA SUCC FUM MAL GLYOXY H2S03 H2S 6-P-Gluconate GLC-LAC RIB-5P RIBO-5P XYL-5P S7P E- 4P MET NADP NADPH acetyl-phosphate ACETAT H-CoA FAD FADH ADP NADH NAD MTHF THF GDP GTP External: BIOMASS GLC METex 02 NH3 C02 S04 GLYCINE Reaction stoichiometry: RI: PEP + GLC = PYR + G6P. R2r: G6P = F6P. R3: G6P + NADP = GLC-LAC + NADPH. R4: GLC-LAC = 6-P-Gluconate. R5: 6-P-Gluconate + NADP = RIB-5P + C02 + NADPH. R6r: RIB-5P = XYL-5P. R7r: RIB-5P = RIBO-5P. R8r: S7P + GA3P = RIBO-5P + XYL-5P. R9r: S7P + GA3P = E-4P + F6P. RIOr: F6P + GA3P = E-4P + XYL-5P. Rll: ATP + F6P = ADP + F-16-BP. R12: F-16-BP = F6P. R13r: F-16-BP = GA3P + DAHP. R14r: DAHP = GA3P. R15r: GA3P + NAD = 13-PG + NADH. R16: ADP + 13-PG = ATP + 3-PG. R17r: 3-PG = 2-PG. R18r: 2-PG = PEP. R19: PEP + ADP = PYR + ATP. R20: PYR + H-CoA + NAD = AC-CoA + NADH + C02.

R21: AC-CoA + OAA = CIT + H-CoA. R22r: CIT = Cis-ACO. R23r: Cis-ACO = ICI. R24: ICI + NADP = 2-OXO + C02 + NADPH. R25: 2-0X0 + NH3 + NADPH = GLU + NADP. R26: 2-OXO + NAD + H-CoA = SUCC-CoA + NADH + C02 R27: SUCC-CoA + GDP = SUCC + H-CoA + GTP. R28r: SUCC + FAD = FUM + FADH. R29r: FUM = BAD. R30r: MAL + NAD = OAA + NADH. R31: ICI = GLYOXY + SUCC. R32: GLYOXY + AC-CoA = MAL + H-CoA. R33: PYR + ATP + C02 = OAA + ADP. R34: PEP + C02 = OAA. R35: OAA + ATP = PEP + ADP + C02. R36: OAA + ADP = PYR + C02 + ATP. R37r: OAA + GLU + NADPH = ASP + 2-0X0 + NADP.

R38: THF + SER = MTHF + GLYCINE. R39: ASP-SA + NADPH = HOM + NADP. R40: HOM + AC-CoA = O-AC-HOM + H-CoA. R41r: 3-PG + NAD = 3-PHP + NADH. R42r: 3-PHP + GLU = SER-P + 2-OXO. R43: SER-P = BE. R44: SER + AC-CoA = O-AC-SER + H-CoA. R45: O-AC-SER + H2S = CYS + ACETAT.

R46: CYS + O-AC-HOM = CYSTA + ACETAT. R47: ASP + ATP = ASP-P + ADP. R48: ASP-P + NADPH = ASP-SA + NADP. R49: O-AC-HOM + H2S = HOMOCYS + ACETAT. R50: ATP + ACETAT = ADP + acetyl-phosphate. R51: acetyl-phosphate + H-CoA = AC-CoA. R52: HOMOCYS + MTHF = MET + THF. R53: MET = METex. R54: CYSTA = HOMOCYS + NH3 + PYR. R55: S04 + 2 ATP + NADPH = H2S03 + 2 ADP + NADP. R56: ATP = ADP. R57: MAL + NADP = PYR + C02 + NADPH. R58: H2S03 + 3 NADPH = H2S + 3 NADP. R59: 2 NADH + 02 + 4 ADP = 2 NAD + 4 ATP. R60: 2 FADH + 02 + 2 ADP = 2 FAD + 2 ATP. R61: 6231 NH3 + 233 S04 + 205 G6P + 308 F6P + 879 RIBO-5P + 268 E-4P + 129 GA3P + 1295 3-PG + 652 PEP + 2604 PYR + 3177 AC-CoA + 1680 OAA + 1224 2-OXO + 16429 NADPH = BIOMASS + 16429 NADP + 3177 H-CoA + 1227 C02. R62: ADP + GTP = ATP + GDP.

The model of wild type E. coli - Reactions and Enzymes: Rl: System phospho-transferase R2: G6P-isomerase R3: G6P-DH R4: Lactonase R5: Gluconate-DH R6: Ribose-5-P-epimerase R7: Ribose- 5-P-isomerase R8: Transketolase 1 R9: Transaldolase RIO: Transketolase 2 Rll: Phosphofruct kinase R12: Fructosebisphosphatase R13: Fructosebisphosphate-aldolase R14: Triosyphosphate isomerase R15: 3-phosphoglycerate-Kinase R16: PG-kinase R17: PG-mutase R18: PEP-hydrolase R19: PYR-kinase R20: PYR-DH R21: CIT-synthase R22: ACO-hydrolase R23: ACONITASE R24: Isocitrate-DH R25: Glutamate-DH R26: 2-OXO-DH R27: SUCC-CoA synthase R28: SUCC-DH R29: FUMARASE R30: MAL-DH R31: ICI-lyase R32: MAL-synthase R33: PYR-carboxylase R34: PEP-carboxylase R35: PEP-carboxykinase R36: OAA-decarboxylase R37: ASP-transaminase R38: Synthesis 1 of M-THF R39: HOM-DH R40: HOM-transacetylase R41: PG-DH R42: Phosphoserine transaminase R43: Phosphoserine phosphatase R44: Serine transacetylase R45: Cysteine synthase R46: Cystathionine synthase R47: Aspartokinase R48: ASP-P-DH R50: ACETAT-kinase R51: Phosphotransacetylase R52: MET-synthase (MetE / H) R53: Exporter of methionine R54 Cystathionine-D-lyase R55 ATP-sulfurylase R56 Hydrolysis of ATP R57 Magmatic enzyme R58 Sulfite-reductase R59 Respiratory chain 1 R60 Respiratory chain 2 R61 Formation of biomass R62 GTP -ATP-Phosphotransferase R70 Transhydrogenase R71 Cleavage of glycine 1 R72 Cleavage of glycine 2 Type of reaction (reversible or irreversible): Reversible: R2r R6r R7r R8r R9r RIOr R13r R14r R15r R17r Rl8r R22r R23r R28r R29r R30r R37r R41r R42r R70r Irreversible. Rl R3 R4 R5 R12 R12 R12 R16 R19 R20 R21 R24 R25 R26 R26 R27 R31 R32 R33 R34 R35 R36 R38 R38 R39 R40 R43 R44 R45 R46 R47 R48 R50 R50 R51 R52 R53 R54 R54 R55 R56 R57 R58 R59 R60 R61 R62 R62 R71 R72 Metabolites (internal or external): Internal: G6P F6P F-16-BP ASP ASP-P ASP-SA HOM ATP O-SUCC-HOM HOMOCYS 3-PHP SER-P BE O-AC-SER CYS CYSTA GA3P DAHP 13-PG 3-PG 2-PG AC-CoA PYR PEP CIT OAA CIS-ACO ICI 2-OXO GLU SUCC-CoA SUCC FUM MAL GLYOXY H2S03 H2S 6-P-Gluconate GLC-LAC RIB-5P RIB0-5P XYL-5P S7P E- 4P MET NADP NADPH H-CoA FAD FADH ADP NADH NAD MTHF THF GDP GTP ACETAT Acetyl Phosphate HPL Methyl-HPL GLYCINE Exters: BIOMASS GLC METex 02 NH3 C02 S04 Reaction stoichiometry: RI: PEP + GLC = PYR + G6P. R2r: G6P = F6P. R3: G6P + NADP = GLC-LAC + NADPH. R4: GLC-LAC = 6-P-Gluconate. R5: 6-P-Gluconate + NADP = RIB-5P + C02 + NADPH. R6r: RIB-5P = XYL-5P. R7r: RIB-5P = RIB0-5P. R8r: S7P + GA3P = RIBO-5P + XYL-5P. R9r: S7P + GA3P = E-4P + F6P. RIOr: F6P + GA3P = E-4P + XYL-5P. Rll: ATP + F6P = ADP + F-16-BP. R12: F-16-BP = F6P. R13r: F-16-BP = GA3P + DAHP. R14r: DAHP = GA3P. R15r: GA3P + NAD = 13-PG + NADH.

R16: ADP + 13-PG = ATP + 3-PG. R17r: 3-PG = 2-PG. R18r: 2-PG = PEP. R19: PEP + ADP = PYR + ATP. R20: PYR + H-CoA + NAD = AC-CoA + NADH + C02.

R21: AC-CoA + OAA = CIT + H-CoA. R22r: CIT = Cis-ACO. R23r: Cis-ACO = ICI. R24: ICI + NADP = 2-OXO + C02 + NADPH. R25: 2-OXO + NH3 + NADPH = GLU + NADP. R26: 2-0X0 + NAD + H-CoA = SUCC-CoA + NADH + C02 R27: SUCC-CoA + GDP = SUCC + H-CoA + GTP. R28r: SUCC + FAD = FUM + FADH. R29r: FUM = BAD. R30r: MAL + NAD = OAA + NADH. R31: ICI = GLYOXY + SUCC. R32: GLYOXY + AC-CoA = MAL + H-CoA. R33: PYR + ATP + C02 = OAA + ADP. R34: PEP + C02 = OAA. R35: OAA + ATP = PEP + ADP + C02. R36: OAA + ADP = PYR + C02 + ATP. R37r: OAA + GLU + NADPH = ASP + 2-OXO + NADP.

R38: THF + SER = MTHF + GLYCINE. R39: ASP-SA + NADPH = HOM + NADP. R40: HOM + SUCC-CoA = O-SUCC-HOM + H-CoA.

R41r: 3-PG + NAD-3-PHP + NADH. R42r: 3-PHP + GLU = SER-P + 2-OXO. R43 SER-P = BE. R44 SER + AC-CoA = O-AC-SER + H-CoA. R45 O-AC-SER + H2S = CYS + ACETAT. R46 CYS + O-SUCC-HOM = CYSTA + SUCC. R47 ASP + ATP = ASP-P + ADP. R48 ASP-P + NADPH = ASP-SA + NADP. R52 HOMOCYS + MTHF = MET + THF. R53 MET = METex. R54 CYSTA = HOMOCYS + NH3 + PYR. R55 S04 + 2 ATP + NADPH = H2S03 + 2 ADP + NADP. R56 ATP = ADP. R57 MAL + NADP = PYR + C02 + NADPH. R58 H2S03 + 3 NADPH = H2S + 3 NADP. R59 2 NADH + 02 + 4 ADP = 2 NAD + 4 ATP. R60 2 FADH + 02 + 2 ADP = 2 FAD + 2 ATP. R61 6965 NH3 + 233 S04 + 206 G6P + 72 F6P + 627 RIBO-5P + 361 E-4P + 129 GA3P + 1338 3-PG + 720 PEP + 2861 PYR + 2930 AC -CoA + 1481 OAA + 1078 2-OXO + 16548 NADPH = BIOMASS + 16543 NADP + 2930 H-CoA + 1678 C02. R62 ADP + GTP = ATP + GDP. R50 ATP + ACETAT = ADP + acetyl-phosphate. R51 acetyl-phosphate + H-CoA = AC-CoA. R70r: NADPH + NAD = NADP + NADH.

R71: GLYCINE + HPL = Methyl-HPL + C02. R72: Methyl-HPL + THF = HPL + MTHF + NH3

Claims (22)

1. CLAIMS 1. A method for determining an organism with increased efficiency for methionine synthesis, wherein the method comprises the steps of a. assigning parameters, by means of a plurality of parameters, the metabolic flux of an initial organism synthesizing methionine based on metabolic pathways already known in relation to methionine synthesis; b. determining a theoretical model of an organism with increased efficiency for methionine synthesis by modifying at least one of the plurality of parameters and / or introducing at least one additional parameter such that the efficiency of methionine synthesis is increased as compared to the initial organism synthesizer of methionine.
2. 2. A device for determining an organism with increased efficiency for methionine synthesis, the device comprises a processor adapted to carry out the following steps of method a. assign parameters, by means of a plurality of parameters, the metabolic flux of an initial wild type organism methionine synthesizer based on metabolic pathways already known in relation to methionine synthesis; b. determining a theoretical model of an organism with increased efficiency for methionine synthesis by modifying at least one of the plurality of parameters and / or introducing at least one additional parameter such that the efficiency of methionine synthesis is increased as compared to the initial organism synthesizer of methionine.
3. 3. A computer-readable medium, in which a computer program for determining an organism with increased efficiency for methionine synthesis is stored which, when being executed by a processor, is adapted to carry out the following stages of the method a. assigning parameters, by means of a plurality of parameters, the metabolic flux of an initial organism synthesizing methionine based on metabolic pathways already known in relation to methionine synthesis; b. determining a theoretical model of an organism with increased efficiency for methionine synthesis by modifying at least one of the plurality of parameters and / or introducing at least one additional parameter such that the efficiency of methionine synthesis is increased as compared to the initial organism synthesizer of methionine. .
4. A program element for determining an organism with increased efficiency for methionine synthesis which, when being executed by a processor, is adapted to carry out the following steps of the method. to. assigning parameters, by means of a plurality of parameters, the metabolic flux of an initial organism synthesizing methionine based on metabolic pathways already known in relation to methionine synthesis; b. determining a theoretical model of an organism with increased efficiency for methionine synthesis by modifying at least one of the plurality of parameters and / or introducing at least one additional parameter such that the efficiency of methionine synthesis is increased as compared to the initial organism synthesizer of methionine.
5. 5. A method for producing an organism that is selected from the group of prokaryotes, lower eukaryotes and plants with increased efficiency of methionine synthesis compared to the starting organisms comprising the following steps: a. assigning parameters, by means of a plurality of parameters, the metabolic flux of an initial organism synthesizing methionine based on metabolic pathways already known in relation to methionine synthesis; b. determining a theoretical model of an organism with increased efficiency for methionine synthesis by modifying at least one of the plurality of parameters and / or introducing at least one additional parameter such that the efficiency of methionine synthesis is increased as compared to the initial organism synthesizer of methionine. c. genetically modify a starting organism in such a way that at least one existing metabolic pathway in the organisms is modified so that the metabolic flow of the organism approaches the theoretical model of the organism and / or d. genetically modifying a starting organism in such a way that at least one exogenous metabolic pathway is introduced into the organisms so that the metabolic flux of the organism approaches the theoretical model of the organism and / or e. provide at least some external metabolites in an amount sufficient to channel the metabolic flow through the metabolic pathways, modified in step c and / or introduced in step d.
6. The method according to claim 5, wherein the metabolic flux through at least one of the existing metabolic pathways selected from the group consisting of phosphotransferase system (PTS) via the pentose phosphate (PPP) glycolysis ( EMP) cycle of tricarboxylic acids (TCA) deviation of glyoxylate (GS) anaplerosis (AP) respiratory chain (RC) sulfur assimilation (SA) synthesis of methionine (MS) synthesis of serine / cysteine / glycine (SCGS) system of excision of glycine (GCS) conversion of transhydrogenase (THGC) via 1 (Pl) via 2 (P2) via 3 (P3) via 4 (P4) via 5 (P5) via 6 (P6) via 7 (P7) via 8 (P8) ) is modified by genetic modification of organisms, and / or metabolic fl ux through at least one of the exogenous metabolic pathways selected from the group consisting of glycine cleavage system (GCS) transhydrogenase conversion (THGC) ) Thiosulfate Reductase System (TRS) System Sulfite Reductase (SRS) Sulfate Reductase System (SARS) Formate Converter System (FCS) Methanethiol Converter System (MCS) is introduced by genetic modification of organisms, and / or organisms are cultured in the presence of selected external metabolites of the group consisting of sulfite sulphite sulphide thiosulfate Cl metabolites such as formate, formaldehyde, methanol, methanethiol or its dimethyldisulfide dimer
7. 7. A method for producing an organism that is selected from the group of prokaryotes, lower eukaryotes and plants with increased efficiency of methionine synthesis in comparison with the starting organisms that comprises the following stages: a. modify the metabolic flux through at least one of the metabolic pathways selected from the group consisting of: phosphotransferase system (PTS) via pentose phosphate (PPP) glycolysis (EMP) tricarboxylic acid (TCA) cycle glyoxylate (GS) anaplerosis (AP) respiratory chain (RC) sulfur assimilation (SA) methionine synthesis (MS) serine / cysteine / glycine synthesis (SCGS) glycine cleavage system (GCS) transhydrogenase conversion (THGC) via 1 (Pl) via 2 (P2) via 3 (P3) via 4 (P4) via 5 (P5) via 6 (P6) via 7 (P7) via 8 (P8) through the genetic modification of the organism, and / or b . introduce a metabolic flux through at least one of the exogenous metabolic pathways selected from the group consisting of glycine cleavage system (GCS) transhydrogenase conversion (THGC) Thiosulfate Reductase System (TRS) Sulfite Reductase System (SRS) Sulphate Reductase System (SRS) Formate Converter System (FCS) Methanethiol Converter System (MCS) by genetic modification of the organism, and / or c. culturing the organisms in the presence of at least one external metabolite selected from the group consisting of: sulfate sulphite sulphide thiosulfate organic sources of sulfur Cl metabolites such as formate, formaldehyde, methanol, methanethiol or its dimethyldisulfide dimer.
8. 8. An organism that is selected from the group of prokaryotes, lower eukaryotes and plants with increased efficiency of methionine synthesis compared to the starting organisms obtainable by the methods of any of claims 5 to 7.
9. 9. An organism of according to claim 8 wherein the organism is selected from the group consisting of microorganisms of the genus Coryneba cterium, of the genus Breviba cteri um, of the genus Escheri chia, yeasts and plants.
10. 10. A method for producing a microorganism of the genus Corynebacterium with increased efficiency of methionine production comprising the following steps a. increase and / or introduce metabolic flux through at least one of the selected pathways of the group consisting of: phosphotransferase system (PTS) and / or pentose pathway phosphate (PPP) and / or sulfur assimilation (SA) and / or anaplerosis (AP) and / or methionine synthesis (MS) and / or serine-glycine synthesis (SCGS) and / or glycine cleavage system (GCS) ) and / or conversion of transhydrogenase (THGC) and / or pathway 1 (Pl) and / or pathway 2 (P2) and / or Thiosulfate Reductase System (TRS) and / or Sulfite Reductase System (SRS) and / or System of Sulphate Reductase (SARS) and / or Formate Converter System (FCS) and / or Methanethiol Converter System (MCS) and / or by the genetic modification of the organism compared to the starting organism, and / or b. decrease at least partially the metabolic flux through at least one of the routes selected from the group consisting of: glycolysis (EMP) and / or tricarboxylic acid cycle (TCA) and / or glyoxylate deviation (GS) and / or respiratory chain (RC) and / or R19 and / or R35 and / or R79 and / or via 3 (P3) and / or via 4 (P4) and / or via 7 (P7) and / or by genetic modification of the organism compared to the beginning.
11. The method according to claim 10 wherein the amount and / or activity of the enzymes selected from the group consisting of: Rl in order to produce more G6P and / or R3 in order to produce more GLC-LAC and / or R4 in order to produce more 6-P-Gluconate and / or R5 in order to produce more RIB-5P and / or R6 in order to produce more XYL-5P and / or R7 in order to produce more RIBO-5P and / or R8 in order to produce more S7P and GA3P and / or R9 in order to produce more E-4p and F6P and / or RIO in order to produce more F6P and GA3P and / or R2 with the In order to produce more G6P and / or R55 in order to produce more H2S03 and / or R58 in order to produce more H2S and / or R71 in order to produce more M-HPL and / or R72 in order to produce more Methylene-THF and / or R70 in order to produce more NADPH and / or R81 in order to produce more NADPH and / or R25 in order to produce more Glu and / or R33 and / or R36 in order to produce more OAA and / or R30 in order to produce more MAL and / or R57 in order to produce more Pyr and / or R73 in order to metabolize thiosulfate to sulfide and sulphite and / or R82 in order to import more external thiosulfate into the cell and / or R74 in order to metabolize sulfite to sulfide and / or R75 in order to produce more 10-formyl-THF and / or R76 in order to produce more Methylene-THF and / or R78 in order to produce more Methyl-THF and / or R77 for the purpose of methyl-sulfhydryl O Acetyl-homoserine with methanethiol and / or R80 for the purpose of metabolizing sulphate to sulphite and / or R47 and / or R48 and / or R39 and / or R46 and / or R49 and / or R52 and / or R52 and / or R54 it is increased and / or introduced in comparison with the starting organism, and / or the amount and / or activity of the enzymes selected from the group consisting of Rll in order to produce less F-1,6-BP and / or R13 in order to produce less DHAP and GA3P and / or R14 in order to produce less GA3P and / or R15 in order to produce less 1,3-PG and / or R16 in order to produce less 3-PG and / or R17 in order to produce less 2-PG and / or R18 in order to produce less PEP and / or R19 in order to produce less Pyr and / or R20 in order to produce less Ac-CoA and / or R21 in order to produce less CIT and / or R22 in order to produce less Cis-ACO and / or R 23 in order to produce less ICI and / or R24 in order to produce less 2-OXO and / or R26 in order to produce less SUCC-CoA and / or R27 in order to produce less SUCC and / or R28 with in order to produce less FUM and / or R29 in order to produce less MAL and / or R30 in order to produce less OAA and / or R21 in order to produce less CIT and / or R22 in order to produce less CIS -ACO and / or R23 in order to produce less ICI and / or R31 in order to produce less GLYOXY and SUCC and / or R32 in order to produce less MAL and / or R28 in order to produce less FUM and / or R29 in order to produce less MAL and / or R30 in order to produce less OAA and / or R60 and / or R56 and / or R62 and / or R61 and / or R19 and / or R35 and / or R79 is reduced / reduce at least partially compared to the starter organism.
12. The method according to claim 11 wherein the amount and / or activity of the enzymes selected from the group consisting of: R3 in order to produce more GLC-LAC and / or R4 in order to produce more 6- P-Gluconate and / or R5 in order to produce more RIB-5P and / or RIO in order to produce more F6P and GA3P and / or R2 in order to produce more G6P and / or R55 in order to produce more H2S03 and / or R58 in order to produce more HS and / or R71 in order to produce more M-HPL and / or R72 in order to produce more Methylene-THF and / or R70 in order to produce more NADPH and / or R81 in order to produce more NADPH and / or R25 in order to produce more Glu and / or R33 and / or R36 in order to produce more OAA and / or R30 in order to produce more MAL and / or R57 in order to produce more Pyr and / or R73 in order to metabolize thiosulfate to sulfide and sulphite and / or R82 in order to import more external thiosulfate into the cell and / or R75 in order to produce 10-formyl- THF and / or R76 with the In order to produce more Methylene-THF and / or R78 in order to produce more Methyl-THF and / or R77 for the purpose of methyl-sulfhydrylating O-Acetyl-homoserine with methanethiol and / or R47 and / or R48 and / or R39 and / or R46 and / or R49 and / or R52 and / or R52 and / or R54 and / or R50 for the purpose of metabolizing sulfate to sulfite are increased and / or introduced compared to the starting organism, and / or the amount and / or activity of the enzymes selected from the group consisting of: Rll in order to produce less F-1,6-BP and / or R19 in order to produce less Pyr and / or R20 in order to produce less Ac- CoA and / or R21 in order to produce less CIT and / or R24 in order to produce less 2-OXO and / or R26 in order to produce less SUCC-CoA and / or R27 in order to produce less SUCC and / or R31 in order to produce less GLYOXY and SUCC and / or R32 in order to produce less MAL and / or R19 in order to produce less Pyruvate and / or R35 in order to produce less PEP and / or R79 with in order to produce less THF are reduced by the m partially in comparison with the starting organism.
13. The method of claim 11 wherein the amount and / or activity of the enzymes selected from the group consisting of R3 in order to produce more GLC-LAC and / or R4 in order to produce more 6-P-Gluconate and / or R5 in order to produce more RIB-5P and / or RIO in order to produce more F6P and GA3P and / or R2 in order to produce plus G6P and R55 in order to produce more H2S03 and / or R58 in order to produce more HS and R71 in order to produce more M-HPL and / or R72 in order to produce more Methylene-THF and / or R78 in order to produce more Methyl-THF and R70 in order to produce more NADPH and / or R81 in order to produce more NADPH and / or R25 in order to produce more Glu and / or R33 and / or R36 with the In order to produce more OAA and / or R30 in order to produce more MAL and / or R57 in order to produce more Pyr and / or R73 in order to metabolize thiosulfate to sulfur and sulphite and R82 in order to import more thiosulfate external to the cell and / or R75 in order to produce 10-formyl-THF and / or R76 in order to produce Methylene-THF and R77 for the purpose of methyl-sulfhydrylated O-Acetyl-homoserine with methanethiol and / or R47 and / or R48 and / or R39 and / or R46 and / or or R49 and / or R52 and / or R52 and / or R54 and / or R80 in order to metabolize sulphate to sulfite are increased and / or introduced compared to the starting organism, and / or: the amount and / or activity of the enzymes selected from the group consisting of: Rll in order to produce less F-1,6-BP and / or R19 in order to produce less Pyr and / or R20 in order to produce less Ac-CoA and / or R21 in order to produce less CIT and / or R24 in order to produce less 2-OXO and / or R26 in order to produce less SUCC-CoA and / or R27 in order to produce less SUCC and / or R31 in order to produce less GLYOXY and SUCC and / or R32 in order to produce less MAL and R19 in order to produce less Pyruvate and R35 in order to produce less PEP and R79 in order to produce less THF are reduced at least partially compared to the starting organism.
14. 14. A microorganism of the genus Coryneba cterium which can be obtained by any of the methods according to claims 10 to 13 preferably selected from the group consisting of Coryneba cterium um to ketoacidophilum, C. a cetoglum utami cum, C. acetophilum , C. ammoniagenes, C. gl utamicum, C. lilium, C. ni trilophilus or C. spec, and preferably Corynebacterium gummicum ATCC 13032, Corynebacterium acetoglutamicum ATCC 15806, Corynejacteriuzn a cetoa cidophilum ATCC 13870, Corynebacterium thermoaminogenes FERM BP-1539, Corynebacterium melassecola ATCC 17965, Corynebacterium glutamine cum KFCC10065 or Corynejacterium glutamicum ATCC21608 and Corynebacterium glutamine cum DSM 17322.
15. 15. A method for producing a microorganism of the genus Escheri chia with increased production efficiency of methionine comprising the following stages increase and / or enter the metabolic flow through at least one of the selected routes of the group that consists of: phosphotransferase system (PTS) and / or glycolysis (EMP) and / or tricarboxylic acid cycle (TCA) and / or glyoxylate deviation (GS) and / or via 1 (Pl) and / or sulfur assimilation (SA) and / or anaplerosis (AP) and / or synthesis of methionine (MS) and / or serine / cysteine / glycine (SCGS) and / or glycine cleavage system (GCS) and / or transhydrogenase conversion (THGC) and / or Thiosulfate Reductase System (TRS) and / or Sulfite Reductase System (SRS) and / or Sulfate Reductase System (SARS) and / or Formate Converter System (FCS) and / or Methane Thiol Converting System (MCS) and / or Synthesis of serine / cysteine / glycine (SCGS) compared to the start by the genetic modification of the organism, and / or at least partially diminishing the metabolic flux through at least one of the selected routes of the group that consists of: via pentose phosphate (PPP) and / or R19 in order to produce less Pyruvate and / or R35 in order to produce less PEP and / or R79 with in order to produce less THF via 3 (P3) and / or via 4 (P4) and / or via 7 (P7) compared to the start by the genetic modification of the organism.
16. The method according to claim 15 wherein the amount and / or activity of the enzymes selected from the group consisting of: Rl in order to produce more G6P R2 in order to produce more F6P and / or Rll with the In order to produce more F-1,6-BP and / or R13 in order to produce more DHAP and GA3P and / or R14 in order to produce more GA3P and / or R15 in order to produce more 1,3-PG and / or R16 in order to produce more 3-PG and / or R17 in order to produce more 2-PG and / or R18 in order to produce more PEP and / or R19 in order to produce more Pyr and / or or R20 in order to produce more Ac-CoA and / or R21 in order to produce more CIT and / or R22 in order to produce more Cis-ACO and / or R23 in order to produce more ICI and / or R24 in order to produce more 2-OXO and / or R26 in order to produce more SUCC-CoA and / or R27 in order to produce more SUCC and / or R28 in order to produce more FUM and / or R29 with the in order to produce more MAL and / or R30 in order to produce more OAA and / or R21 with in order to produce more CIT and / or R22 in order to produce more Cis-ACO and / or R23 in order to produce more ICI and / or R31 in order to produce more GLYOXY and SUCC and / or R32 in order to produce more NAL and / or R28 in order to produce more FUM and / or R29 in order to produce more MAL and / or R30 in order to produce more OAA and / or R25 in order to produce more Glu and / or R55 with in order to produce more H S03 and / or R58 in order to produce more H2S and / or R71 in order to produce more M-HPL and / or R72 in order to produce more Methylene-THF and / or R78 with the In order to produce more Methyl-THF and / or R70 in order to produce more NADPH and / or R81 in order to produce more NADPH and / or R73 in order to metabolize thiosulfate to sulfur and sulphite and / or R82 in order of importing more external thiosulfate into the cell and / or R74 in order to metabolize sulfite to sulfide and / or R75 in order to produce more 10-formyl-THF and / or R76 in order to produce more Methylene-THF from 10-formyl-THF and / or R77 for the purpose of methyl-sulfhydrylar O-Acetyl-homoserine with methanothiol and / or R80 in order to metabolize sulfate to sulfite and / or R44 in order to produce more O-Ac-SER and / or R45 in order to produce more CYS is increased and / or introduced compared to the organism of initiation, and / or the amount and / or activity of the enzymes selected from the group consisting of: R3 in order to produce less GLC-LAC and / or R4 in order to produce less 6-P-Gluconate and / or R5 in order to produce less RIB-5P and / or R6 in order to produce less XYL-5P and / or R7 in order to produce less RIBO-5P and / or R8 in order to produce less S7P and GA3P and / or R9 in order to produce less E-4p and F6P and / or RIO in order to produce less F6P and GA3P and / or R2 in order to produce less G6P and / or R49 in order to produce less HOMOCYS and / or R19 in order to produce less Pyruvate and / or R35 in order to produce less PEP and / or R79 in order to produce less THF and / or R56 and / or R62 and / or R61 is reduced / reduced at least partially compared to the starting organism .
17. 17. The method according to claim 16 wherein the amount and / or activity of the enzymes selected from the group consisting of: R1 in order to produce more G6P and / or R2 in order to produce more F6P and / or Rll in order to produce more F-1,6-BP and / or R19 in order to produce more Pyr and / or R20 in order to produce more Ac-CoA and / or R21 in order to produce more CIT and / or R24 in order to produce more 2-OXO and / or R26 in order to produce more SUCC-CoA and / or R31 in order to produce more GLYOXY and SUCC and / or R32 in order to produce more MAL and / or R25 in order to produce more Glu and / or R55 in order to produce more H2S03 and / or R58 in order to produce more H2S and / or R71 in order to produce more M-HPL and / or R72 with in order to produce more Methylene-THF and / or R78 in order to produce more Methyl-THF and / or R70 in order to produce more NADPH and / or R81 in order to produce more NADPH and / or R73 in order of metabolizing thiosulfate to sulfur and sulphite and / or R82 in order to import more external thiosulfate into the cell and / or R74 in order to metabolize sulfite to sulfide and / or R75 in order to produce more 10-formyl-THF and / or R76 in order to produce more Methylene-THF and / or R77 for the purpose of methyl-sulfhydrylating O-Acetyl-homoserine with methanethiol and / or R80 to metabolize sulfate towards sulfite and / or R44 in order to produce more O-Ac-SER and / or R45 in order to produce more CYS is increased / increased and / or introduced compared to the starting organism, and / or the amount and / or activity of the enzymes selected from the group consisting of: R3 in order to produce less GLC-LAC and / or R4 in order to produce less 6-P-Gluconate and / or R5 in order to produce less RIB- 5P and / or RIO in order to produce less F6P and GA3P and / or R19 in order to produce less Pyruvate and / or R35 in order to produce less PEP and / or R79 in order to produce less THF is reduced / they reduce at least partially in comparison with the starting organism
18. 18. A microorganism of the genus Escherichia that can be obtained by any of the methods according to the invention. 15 to 17 preferably selected from the group consisting of E. coli.
19. 19. An organism according to claims 8, 9, 14 or 18 characterized in that the methionine is produced with a molar ratio of methionine to glucose supply of at least 10%, of at least 20%, at less 30%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80% or at least 85%.
20. 20. The use of any of the organisms of claims 8, 9, 14, 18 or 19 to produce methionine.
21. 21. A method for producing methionine comprising the following steps: a. cultivating an organism according to claim 8, 9, 14, 18 or 19 b. isolate methionine
22. 22. The method according to claim 21 wherein the cultivation is ced out in a suitable medium and optionally thiosulfate, sulfite, sulfide and / or Cl compounds such as formate or methanethiol. SUMMARY OF THE INVENTION The present invention concerns methods for the production of microorganisms with increased efficiency for methionine synthesis. The present invention also concerns microorganisms with increased efficiency for methionine synthesis. Additionally, the present invention concerns methods for determining the optimal metabolic flux for organisms with respect to methionine synthesis.