Nothing Special   »   [go: up one dir, main page]

CN1371418A - Genes from corynebacterium glutamicum for the biosynthesis of folic acid and their use for the microbia production of folic acid - Google Patents

Genes from corynebacterium glutamicum for the biosynthesis of folic acid and their use for the microbia production of folic acid Download PDF

Info

Publication number
CN1371418A
CN1371418A CN00812014A CN00812014A CN1371418A CN 1371418 A CN1371418 A CN 1371418A CN 00812014 A CN00812014 A CN 00812014A CN 00812014 A CN00812014 A CN 00812014A CN 1371418 A CN1371418 A CN 1371418A
Authority
CN
China
Prior art keywords
polypeptide
folic acid
sequence
amino acid
corynebacterium glutamicum
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Pending
Application number
CN00812014A
Other languages
Chinese (zh)
Inventor
M·马克
K·赫尔布斯特
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
ACKERSARON BIOSCIENCE AG
Original Assignee
ACKERSARON BIOSCIENCE AG
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by ACKERSARON BIOSCIENCE AG filed Critical ACKERSARON BIOSCIENCE AG
Publication of CN1371418A publication Critical patent/CN1371418A/en
Pending legal-status Critical Current

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/195—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from bacteria
    • C07K14/34—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from bacteria from Corynebacterium (G)
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/10—Transferases (2.)
    • C12N9/1085—Transferases (2.) transferring alkyl or aryl groups other than methyl groups (2.5)
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
    • C12N15/52—Genes encoding for enzymes or proenzymes
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/10—Transferases (2.)
    • C12N9/12—Transferases (2.) transferring phosphorus containing groups, e.g. kinases (2.7)
    • C12N9/1235—Diphosphotransferases (2.7.6)
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/14—Hydrolases (3)
    • C12N9/78—Hydrolases (3) acting on carbon to nitrogen bonds other than peptide bonds (3.5)
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/88—Lyases (4.)
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P17/00—Preparation of heterocyclic carbon compounds with only O, N, S, Se or Te as ring hetero atoms
    • C12P17/18—Preparation of heterocyclic carbon compounds with only O, N, S, Se or Te as ring hetero atoms containing at least two hetero rings condensed among themselves or condensed with a common carbocyclic ring system, e.g. rifamycin
    • C12P17/182—Heterocyclic compounds containing nitrogen atoms as the only ring heteroatoms in the condensed system

Landscapes

  • Chemical & Material Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Organic Chemistry (AREA)
  • Engineering & Computer Science (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Biomedical Technology (AREA)
  • Molecular Biology (AREA)
  • Biochemistry (AREA)
  • General Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Biotechnology (AREA)
  • Microbiology (AREA)
  • Medicinal Chemistry (AREA)
  • Biophysics (AREA)
  • Physics & Mathematics (AREA)
  • Plant Pathology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)

Abstract

The invention relates to nucleotide sequences of four genes (folE, folP, folB and folK) from Corynebacterium glutamicum for the biosynthesis of folic acid and their use for the microbial production of folic acid.

Description

From the folic acid biological synthetic gene of corynebacterium glutamicum and in the application aspect the folic acid microorganisms producing
The present invention relates to utilize the method that produces folic acid through the biology of genetic manipulation by fermentation.The present invention includes four kinds from the nucleotide sequence of the folic acid biological synthetic gene (folE, folP, folB and folK) of corynebacterium glutamicum (Cory nebacterium glutamicum) and in the application aspect the folic acid microorganisms producing.These four kinds of a kind of operons of genomic constitution, and following sequence is transcribed: folE, folP, folB and folK.
Folic acid is the essential composition of animal body.Its derivative tetrahydrofolic acid (THFA) is the first carrier (Carrier von aktivierten Einkohlenstoffeinheiten) of the active very widely single carbon of a kind of purposes in the zooblast.Folic acid contains three kinds of groups: the pteridine ring of replacement, p-benzaminic acid ester group and glutamate group.Mammals can not be synthesized pteridine ring.But can absorb folic acid by diet and enteric microorganism thereof.Folic acid deficiency mainly causes mucosal lesion.
The commercial value of folic acid is animal-feed and human food prods's market segment.Folic acid mainly can be used as a kind of foodstuff additive.
The fermentative production of folic acid can be used microorganism.By can optimize the folic acid biological combined coefficient of microorganism to the genetic manipulation of folic acid biological route of synthesis.Genetic manipulation herein is meant copy number and/or the transcription rate that improves folic acid biological route of synthesis genes involved.Consequently improve the ratio of gene product, thereby improve the enzymic activity in the born of the same parents.The raising of enzymic activity can cause food (as glucose) to improve to the transformation efficiency of folic acid, thereby also causes the raising of production concentration.In order to carry out genetic manipulation, must identify the nucleotide sequence of folic acid biological route of synthesis genes involved.The present invention relates to from corynebacterium glutamicum and be used for four kinds of new gene orders of folic acid biological synthetic and in the application aspect the folic acid microorganisms producing.
On the one hand, the present invention includes the folE gene product.Sequence numbering: 2 show a kind of peptide sequence.A kind of molecular weight of being made up of 202 amino acid of folE gene product coding is the polypeptide of 22029Da.The invention still further relates to the functional derivatives of this polypeptide, these derivatives can make up sequence numbering by certain that utilizes disappearance, insertion or replacement or disappearance, inserts and replace: one or more amino acid of 2, preferably reach 25% amino acid, optimal then is to reach 15% amino acid, and the method for being replaced obtains.Term " functional derivatives " is meant enzymic activity and contains sequence numbering: polypeptide of sequence shown in 2 still is in the derivative of same order.
On the other hand, the present invention includes the folP gene product.Sequence numbering: 4 show a kind of peptide sequence.A kind of molecular weight of being made up of 285 amino acid of folP gene product coding is the polypeptide of 29520Da.The invention still further relates to the functional derivatives of this polypeptide, these derivatives can make up sequence numbering by certain that utilizes disappearance, insertion or replacement or disappearance, inserts and replace: one or more amino acid of 4, preferably reach 40% amino acid, optimal then is to reach 25% amino acid, and the method for being replaced obtains.Term " functional derivatives " is meant enzymic activity and contains sequence numbering: polypeptide of sequence shown in 4 still is in the derivative of same order.
On the other hand, the present invention includes the folB gene product.Sequence numbering: 6 show a kind of peptide sequence.A kind of molecular weight of being made up of 131 amino acid of folB gene product coding is the polypeptide of 14020Da.The invention still further relates to the functional derivatives of this polypeptide, these derivatives can make up sequence numbering by certain that utilizes disappearance, insertion or replacement or disappearance, inserts and replace: one or more amino acid of 6, preferably reach 30% amino acid, optimal then is to reach 20% amino acid, and the method for being replaced obtains.Term " functional derivatives " is meant enzymic activity and contains sequence numbering: polypeptide of sequence shown in 6 still is in the derivative of same order.
On the other hand, the present invention includes the folK gene product.Sequence numbering: 8 show a kind of peptide sequence.A kind of molecular weight of being made up of 160 amino acid of folk gene product coding is the polypeptide of 18043Da.The invention still further relates to the functional derivatives of this polypeptide, these derivatives can make up sequence numbering by certain that utilizes disappearance, insertion or replacement or disappearance, inserts and replace: one or more amino acid of 8, preferably reach 40% amino acid, optimal then is to reach 30% amino acid, and the method for being replaced obtains.Term " functional derivatives " is meant enzymic activity and contains sequence numbering: polypeptide of sequence shown in 8 still is in the derivative of same order.
On the other hand, the present invention includes the polynucleotide sequence of codified aforementioned polypeptides.The generation of these polynucleotide sequences can (be a sequence numbering: 1,3,5 and 7) beginning by the sequence of separating from corynebacterium glutamicum, by site-directed mutagenesis these sequences are modified, or utilize genetic code corresponding polypeptide to be translated and adopted fully on this basis the method for chemosynthesis.
Preferably, these polynucleotide sequences can be used to transform host living beings, in this microorganism preferably, particularly adopt the form of gene construct, and this type of construct contains a copy of one of these polynucleotide at least, and contains a kind of adjusting sequence at least.Regulate in the sequence and comprise promotor, terminator, enhanser and ribosome bind site.
The preferred host living beings that utilizes these gene constructs to transform is corynebacterium (Corynebacterium) and bacillus (Bacillus) species.Also can adopt any eukaryotic microorganisms, preferably Ash is than the yeast strain of yeast belong (Ashbya), mycocandida (Candida), Pichia (Pichia), yeast belong (Sac charomyces) and Hansenula (Hansenula).
On the other hand, the present invention includes the production method of folic acid, this method comprises is cultivated host transformed biology in the above described manner, separates folic acid then.
Culturing micro-organisms and understood by those of skill in the art by the method and the process of separating folic acid in the microniological proudcts.
Following examples will be used for the microorganism hereditary operation to the present invention and describe to improve should being used as in more detail of folic acid combined coefficient.
The structure of embodiment 1 corynebacterium glutamicum ATCC 13032 genomic libraries
Utilize the described ordinary method of forefathers, (1984, method Mol.Gen.Genet.195:134-138) can obtain the genomic dna of corynebacterium glutamicum ATCC 13032 as J.Altenbuchner and J.Cullum.According to conventional scheme (as Sambrook, J.et al. (1989) molecular cloning: laboratory manual, Cold Spring Harbor LaboratoryPress) utilizes any cloning vector, as pBluescript II KS-(Stratagene) or ZAP Express TM(Stratagene) all can produce genomic library.Can use the fragment of any size in addition, preferably length is the Sau3AI fragment of 2-9kb, and these fragments can merge in the cloning vector of BamHI digestion.
The nucleic acid sequence analysis of embodiment 2 genomic libraries
The genomic library that is made up by embodiment 1 filters out one escherichia coli cloning.Utilize the ordinary method LB of 100mg/l penbritin (as add) in appropriate culture medium to cultivate Bacillus coli cells, separablely then go out plasmid DNA.Be cloned among the pBluescript II KS-(seeing embodiment 1) the corynebacterium glutamicum genomic DNA fragment can by oligonucleotide 5 '-AATTAACCCTCACTAAAGGG-3 ' and 5 '-GTAATACGACTCACTATAGGGC-3 ' checked order.
The Computer Analysis of embodiment 3 isolated nucleic acid sequences
By, for example, BLASTX algorithm (Altschul et al. (1990) J.Mol.Biol.215:403-410) can link together nucleotide sequence.New sequence might be found like this, and the function of these new genes can be understood.
Embodiment 4 contains the evaluation of the escherichia coli cloning of GTP cyclization hydrolase I (EC 3.5.4.16) gene nucleotide series
As described in embodiment 2, escherichia coli cloning analyzed and after its calling sequence being analyzed as described in the embodiment 3, finding one section as sequence numbering: the sequence shown in 1.Utilize BLASTX algorithm (seeing embodiment 3) to find this sequence and GTP cyclization hydrolase I (FolE from different biologies; EC 3.5.4.16) similarity is arranged.Property is the highest similarly is that (NRDB 006273 for GTP cyclization hydrolase I (FolE) from mycobacterium tuberculosis (Mycobacterium tuberculosis); On amino acid levels, have 72% identical).
Embodiment 5 contains the evaluation of the escherichia coli cloning of dihydropteroate synthase (EC 2.5.1.15) gene nucleotide series
As described in embodiment 2, escherichia coli cloning analyzed and after its calling sequence being analyzed as described in the embodiment 3, finding one section as sequence numbering: the sequence shown in 3.Utilize BLASTX algorithm (seeing embodiment 3) to find this sequence and dihydropteroate synthase (FolP from different biologies; EC 2.5.1.15) similarity is arranged.Property is the highest similarly is that (NRDB 006274 for dihydropteroate synthase (FolP) from mycobacterium tuberculosis; On amino acid levels, have 53% identical).
Embodiment 6 contains the evaluation of the escherichia coli cloning of dihydroneopterin aldolase (EC 4.1.2.25) gene nucleotide series
As described in embodiment 2, escherichia coli cloning analyzed and after its calling sequence being analyzed as described in the embodiment 3, finding one section as sequence numbering: the sequence shown in 5.Utilize BLASTX algorithm (seeing embodiment 3) to find this sequence and dihydroneopterin aldolase (FolB from different biologies; EC 4.1.2.25) similarity is arranged.Property is the highest similarly is that (NRDB 006275 for dihydroneopterin aldolase (FolB) from mycobacterium tuberculosis; On amino acid levels, have 61% identical).Embodiment 7 contains the evaluation of the escherichia coli cloning of 2-amino-4-hydroxy-6-hydroxymethyl dihydropteridine pyrophosphokinase (EC 2.7.6.3) gene nucleotide series
As described in embodiment 2, escherichia coli cloning analyzed and after its calling sequence being analyzed as described in the embodiment 3, finding one section as sequence numbering: the sequence shown in 7.Utilize BLASTX algorithm (seeing embodiment 3) to find this sequence and 2-amino-4-hydroxy-6-hydroxymethyl dihydropteridine pyrophosphokinase (FolK from different biologies; EC 2.7.6.3) similarity is arranged.Property is the highest similarly is 2-amino-4-hydroxy-6-hydroxymethyl dihydropteridine pyrophosphokinase (Folk) (EMBL AL023093 from Mycobacterium leprae; On amino acid levels, have 43% identical).
Embodiment 8 is from GTP cyclization hydrolase I, dihydro pterin ester (Dihydrop eroat) synthase, dihydroneopterin aldolase and the 2-amino-4-hydroxy-6-hydroxymethyl dihydropteridine pyrophosphokinase application in folic acid is produced of corynebacterium glutamicum
Utilize suitable clone and expression system the gene from GTP cyclization hydrolase I, dihydropteroate synthase, dihydroneopterin aldolase and the 2-amino-4-hydroxy-6-hydroxymethyl dihydropteridine pyrophosphokinase of corynebacterium glutamicum can be incorporated in corynebacterium glutamicum or other any microorganism.Thereby can be created in activity or gene copy number aspect the microorganism through genetic manipulation different with wild-type biology.These new bacterial strains through genetic manipulation can be used to produce folic acid.
Sequence list
(I) general information (1) applicant
(A) title: BASF-LYNX Bioscience AG
(B) street: Im Neuenheimer Feld 515
(C) city: Heidelberg
(D) country: Germany
(E) postcode: 69120
(F) phone: 06221/4546
(G) fax: 06221/454770 (2) title: the glutamic acid rod that derives from that is used for biosynthesizing folic acid
The gene of shape bacillus and be used for microorganisms producing
( 3 ) ：8 SEQ ID NO.1：DNA ( folE ) ATGAAGGAGACAACCGTGGATAACCACGCTGCAGTTCGCGAGTTCGATGAGGAGCGCGCAACAGCTGCGATTCGTGAGTTGCTCATCGCTGTGGGTGAGGATCCAGATCGCGAAGGCCTGTTGGAAACCCCAGCTCGAGTGGCTAGGGCGTACAAGGAAACTTTCGCGGGTCTGCATGAGGATCCCACCACTGTGCTGGAGAAGACGTTCTCTGAGGGCCATGAAGAGTTGGTTCTGGTTCGTGAGATCCCGATTTACTCCATGTGTGAGCACCACTTGGTGCCGTTCTTTGGCGTGGCGCACATTGGTTACATTCCGGGTAAGTCCGGCAAGGTGACTGGCCTGTCCAAGCTGGCGCGTTTAGCGGATATGTTTGCTAAGCGACCTCAGGTTCAGGAGCGCTTGACCTCCCAAATTGCGGATGCTCTCGTCGAAAAGCTTGATGCCCAGGCCGTGGCCGTGGTGATTGAAGCTGAGCACCTGTGCATGGCCATGCGCGGAATCCGTAAGCCTGGTGCTGTGACCACGACGTCTGCGGTGCGCGGCGGTTTTAAGAACAACGCTGCCTCCCGCGCTGAGGTGTTCTCCCTGATTCGGGGGCACTAA
SEQ ID NO.2: amino acid (FolE) MKETTVDNHAAVREFDEERATAAIRELLIAVGEDPDREGLLETPARVARAYKETFA GLHEDPTTVLEKTFSEGHEELVLVREIPIYSMCEHHLVPFFGVAHIGYIPGKSGKV TGLSKLARLADMFAKRPQVQERLTSQIADALVEKLDAQAVAVVIEAEHLCMAMRGI RKPGAVTTTSAVRGGFKNNAASRAEVFSLIRGH
SEQ?ID?NO.3：DNA(folP)ATGAACGTATCCTCTTTGACCATCCCGGGACGCTGTTTGGTCATGGGAATTGTCAATGTCACTGAGGATTCCTTTTCGGACGGTGGCAAGTACATTGACGTTGATCAGGCGATCGCGCATGCCAAGGAATTGGTGGCTGCTGGCGCCGACATGATTGATGTCGGCGGCGAGTCCACCCGGCCTGGGGCAGTGCGCGTCGACGCGTCCGTGGAACGGGACCGGGTTGTGCCGGTCATTAAGGCGCTTCACGACGCCGGCATCCACACTTCCGTAGACACCATGCGGGCCTCCGTGGCGCAGGCTGCCGCGGGCGCTGGCGTCTCCATGATCAACGACGTCTCTGGCGGTTTGGCTGATCCTGAGATGTTTTCTGTCATGGCGGAAGCGCAAATTCCCGTGTGTTTGATGCACTGGCGCACCCTCCAATTCGGTGATGCCGCAGGTCAGGCAGATCACGGTGGAGACGTTGTAGCCGATGTGCACGCAGTGCTTGATGATCTTGTCGCCCGCGCCACCGCTGCTGGTGTGGCCGAAAACCAGATCGTGCTTGATCCAGGTTTGGGTTTTGCCAAATCACGTGAAGACAACTGGCGTTtGCTGCAAGCACTGCCCGAGTTTATTTCTGGACCTTTCCCCATCCTGGTGGGAGCATCCCGGAAGCGATTCCTGGCTGGCGTGCGCAAAGACCGTGGCCTAGATGTCACCCCCATTGATGCCGACCCAGCAACCGCAGCGGTGACCGCAGTGTCTGCACATATGGGAGCATGGGGTGTGCGCGTGCACGATGTCCCAGTATCAAGGGACGCTGTTGATGTTGCCGCATTGTGGCGAAGTGGAGGAACTCACCATGGCTGA
SEQ ID NO.4: amino acid (FolP) MNVSSLTIPGRCLVMGIVNVTEDSFSDGGKYIDVDQAIAHAKELVAAGADMIDVGG ESTRPGAVRVDASVERDRVVPVIKALHDAGIHTSVDTMRASVAQAAAGAGVSMIND VSGGLADPEMFSVMAEAQIPVCLMHWRTLQFGDAAGQADHGGDVVADVHAVLDDLV ARATAAGVAENQIVLDPGLGFAKSREDNWRLLQALPEFISGPFPILVGASRKRFLA GVRKDRGLDVTPIDADPATAAVTAVSAHMGAWGVRVHDVPVSRDAVDVAALWRSGG THHG
SEQ?ID?NO.5：DNA(folB)ATGGCTGATCGTATTGAACTTAAAGGCCTTGAATGCTTCGGACACCACGGTGTGTTCGACTTTGAAAAAGAGCAAGGCCAGCCCTTCATTGTGGATGTCACCTGCTGGATGGATTTCGATGCCGCAGGTGCCAGCGATGACCTTTCCGACACCGTAGATTACGGCGCGTTGGCATTGTTGGTTGCTGAAATCGTGGAAGGCCCATCCAGGGATTTGATCGAGACGGTGGCCACGGAATCTGCGGATGCTGTGATGGCTAAATTTGATGCGCTTCATGCGGTGGAAGTAACCATCCATAAGCCCAAAGCACCGATCCCACGTACTTTTGCTGACGTCGCGGTGGTTGCCCGACGTTCCAGGAAATCCATGGCTGCTGGAAGGAGCAACGCCTAA
SEQ ID NO.6: amino acid (FolB) MADRIELKGLECFGHHGVFDFEKEQGQPFIVDVTCWMDFDAAGASDDLSDTVDYGA LALLVAEIVEGPSRDLIETVATESADAVMAKFDALHAVEVTIHKPKAPIPRTFADV AVVARRSRKSMAAGRSNA
SEQ?ID?NO.7：DNA(folK)ATGCATGCAGTTTTGTCCATCGGTTCCAACATGGATGATCGCTACGCGCTGCTCAACACAGTGATCGAGGAATTCAAAGATGAGATCGTGGCGCAGTCTGCGATCTACTCAACCCCACCGTGGGGCATTGAGGATCAGGATGAATTCCTCAACGCAGTGCTCGTTGTTGAGGTTGAAGAAACCCCCATCGAGTTGCTGCGCCGTgGCCAAAAACTCGAAGAAGCCGCCGAGCGGGTCCGCGTCCGCAAATGGGGGCCACGCACCCTCGATGTGGATATCGTGCAGATCATTAAAGATGGGGAAGAGATCCTTTCTGAGGATCCCGAACTGACCTTGCCACACCCTTGGGCTTGGCAGCGTGCCTTCGTGTTGATCCCTTGGTTGGAAGCAGAACCTGATGCCGTCCTGCACGGCACGACCATTGCAGAACATGTGGATAATCTTGATCCCACAGACATTGAAGGTGTCACCAAGATTTAA
SEQ ID NO.8: amino acid (FolK) MHAVLSIGSNMDDRYALLNTVIEEFKDEIVAQSAIYSTPPWGIEDQDEFLNAVLVV EVEETPIELLRRGQKLEEAAERVRVRKWGPRTLDVDIVQIIKDGEEILSEDPELTL PHPWAWQRAFVLIPWLEAEPDAVLHGTTIAEHVDNLDPTDIEGVTKI

Claims (8)

1. one kind has the active polypeptide of GTP cyclization hydrolase I, and this polypeptide is selected from:
(a) a kind of have as sequence numbering: the polypeptide of aminoacid sequence as described in 2;
(b) a kind of polypeptide with (a) is compared through one or more amino acid whose disappearances, insertion or replacement and adorned polypeptide.
2. one kind has the active polypeptide of dihydropteroate synthase, and this polypeptide is selected from:
(a) a kind of have as sequence numbering: the polypeptide of aminoacid sequence as described in 4;
(b) a kind of polypeptide with (a) is compared through one or more amino acid whose disappearances, insertion or replacement and adorned polypeptide.
3. one kind has the active polypeptide of dihydroneopterin aldolase, and this polypeptide is selected from:
(a) a kind of have as sequence numbering: the polypeptide of aminoacid sequence as described in 6;
(b) a kind of polypeptide with (a) is compared through one or more amino acid whose disappearances, insertion or replacement and adorned polypeptide.
4. 2-amino-4-hydroxy-active polypeptide of 6-hydroxymethyl dihydropteridine pyrophosphokinase, this polypeptide is selected from:
(a) a kind of have as sequence numbering: the polypeptide of aminoacid sequence as described in 8;
(b) a kind of polypeptide with (a) is compared through one or more amino acid whose disappearances, insertion or replacement and adorned polypeptide.
5. polynucleotide, it is encoded corresponding to claim 1, and 2,3, or 4 polypeptide.
6. gene constructs, it contains the polynucleotide of claim 5 of at least one copy and at least one regulates sequence.
7. host organisms, it is transformed by the gene constructs with claim 6.
8. be used to prepare the method for folic acid, also separate folic acid subsequently by the host organisms of cultivating claim 7.
CN00812014A 1999-06-25 2000-06-23 Genes from corynebacterium glutamicum for the biosynthesis of folic acid and their use for the microbia production of folic acid Pending CN1371418A (en)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
DE19929363.5 1999-06-25
DE19929363A DE19929363A1 (en) 1999-06-25 1999-06-25 New Corynebacterium glutamicum GTP cyclohydrolase I, dihydropteroate synthase, dihydroneopterin aldolase and 2-amino-4-hydroxy-6-hydroxymethyl-dihydropteridine pyrophosphokinase polypeptides

Publications (1)

Publication Number Publication Date
CN1371418A true CN1371418A (en) 2002-09-25

Family

ID=7912677

Family Applications (1)

Application Number Title Priority Date Filing Date
CN00812014A Pending CN1371418A (en) 1999-06-25 2000-06-23 Genes from corynebacterium glutamicum for the biosynthesis of folic acid and their use for the microbia production of folic acid

Country Status (8)

Country Link
EP (1) EP1194565A1 (en)
KR (1) KR20020026469A (en)
CN (1) CN1371418A (en)
AU (1) AU5978200A (en)
CA (1) CA2377458A1 (en)
DE (1) DE19929363A1 (en)
WO (1) WO2001000845A1 (en)
ZA (1) ZA200200582B (en)

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1952114B (en) * 2005-10-20 2010-04-14 浙江爱迪亚营养科技开发有限公司 Glutamic acid corynebacterium and process for preparing nicotinamide by using same
CN109810991A (en) * 2019-03-02 2019-05-28 昆明理工大学 The purposes of dihydropteroate synthase gene folP
CN111235169A (en) * 2020-02-03 2020-06-05 昆明理工大学 GTP cyclohydrolase I gene folE and application thereof
CN112852844A (en) * 2021-03-05 2021-05-28 昆明理工大学 Application of hydroxymethyl dihydropterin pyrophosphokinase gene folK

Families Citing this family (16)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1298854C (en) 1999-07-02 2007-02-07 味之素株式会社 DNA encoding sucrose PTS enzyme II
US6942996B2 (en) 2000-08-02 2005-09-13 Degussa Ag Isolated polynucleotide from Corynebacterium encoding a homocysteine methyltransferase
US6958228B2 (en) 2000-08-02 2005-10-25 Degussa Ag Nucleotide sequence which code for the metH gene
DE10039049A1 (en) 2000-08-10 2002-02-21 Degussa Novel polynucleotide from Coryneform bacteria coding for lysR3 gene, useful as a probe for detecting DNA to isolate nucleic acids coding for transcription regulator lysR3 or for producing L-amino acids, e.g., L-lysine and L-valine
DE10039044A1 (en) 2000-08-10 2002-02-21 Degussa Novel polynucleotide from Coryneform bacteria coding for lysR1 gene, useful as hybridization probe for detecting DNA coding for transcription regulator lysR1
DE10039043A1 (en) 2000-08-10 2002-02-21 Degussa New nucleotide sequences coding for the luxR gene
AU2001291658A1 (en) 2000-08-26 2002-03-13 Degussa A.G. Nucleotide sequences which code for the ccpa2 gene
US6812016B2 (en) 2000-09-02 2004-11-02 Degussa Ag Nucleotide sequences which code for the metY gene
US6815196B2 (en) 2000-09-02 2004-11-09 Degussa Ag Nucleotide sequences encoding o-succinylhomoserine sulfhydrylase
WO2002020792A1 (en) 2000-09-09 2002-03-14 Degussa Ag Efflux protein dep33 of corynebacterium glutamicum
US6759224B2 (en) 2000-09-09 2004-07-06 Degussa Ag Nucleotide sequences which code for the sahH gene
DE10045496A1 (en) 2000-09-14 2002-03-28 Degussa New nucleotide sequences coding for the ptsi gene
DE10055870A1 (en) 2000-11-10 2002-05-29 Degussa New nucleotide sequences coding for the nadC gene
DE10055869A1 (en) 2000-11-10 2002-05-29 Degussa A polynucleotide encoding the nadA gene useful for the preparation of nicotinic acid or its derivatives, as probes for discovering RNA, cDNA and DNA to isolate polynucleotides or genes which code for quinolinate synthetase A
EP1262541A1 (en) * 2001-05-28 2002-12-04 Stichting Top-Instituut Voedselwetenschappen Production of bioavailable folic acid
US7468262B2 (en) 2003-05-16 2008-12-23 Ajinomoto Co., Inc. Polynucleotides encoding useful polypeptides in corynebacterium glutamicum ssp. lactofermentum

Family Cites Families (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5968788A (en) * 1995-08-28 1999-10-19 Toray Industries, Inc. Method for producing folic acid

Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1952114B (en) * 2005-10-20 2010-04-14 浙江爱迪亚营养科技开发有限公司 Glutamic acid corynebacterium and process for preparing nicotinamide by using same
CN109810991A (en) * 2019-03-02 2019-05-28 昆明理工大学 The purposes of dihydropteroate synthase gene folP
CN109810991B (en) * 2019-03-02 2021-11-12 昆明理工大学 Application of dihydropteroate synthase gene folP
CN111235169A (en) * 2020-02-03 2020-06-05 昆明理工大学 GTP cyclohydrolase I gene folE and application thereof
CN112852844A (en) * 2021-03-05 2021-05-28 昆明理工大学 Application of hydroxymethyl dihydropterin pyrophosphokinase gene folK

Also Published As

Publication number Publication date
DE19929363A1 (en) 2000-12-28
KR20020026469A (en) 2002-04-10
ZA200200582B (en) 2003-03-26
CA2377458A1 (en) 2001-01-04
AU5978200A (en) 2001-01-31
EP1194565A1 (en) 2002-04-10
WO2001000845A1 (en) 2001-01-04

Similar Documents

Publication Publication Date Title
CN1371418A (en) Genes from corynebacterium glutamicum for the biosynthesis of folic acid and their use for the microbia production of folic acid
KR102055874B1 (en) A Novel Isopropylmalate Synthase Variant and a Method of Producing L-Leucine Using the Same
CN101960005B (en) The production method of 5'-GMP
Conway et al. Phosphoglycerate kinase gene from Zymomonas mobilis: cloning, sequencing, and localization within the gap operon
CN102021154A (en) Method for improving yield of arginine by mutation of Corynebacterium crenatum N-acetyl glutamic acid kinase
CN109415418A (en) The method for generating interested molecule by the inclusion of the microbial fermentation of the gene of coding sugar phosphotransferase system (PTS)
AU5685400A (en) Dihydroorotate dehydrogenase sequence of corynebacterium glutamicum and the use thereof in microbial production of pyrimidine and/or compounds used with pyrimidine
CN104480169A (en) Method for producing 5'-guanylic acid
TW201793B (en)
US5281531A (en) Host and vector for producing D-ribose
JP4216719B2 (en) Halogen compound-resistant novel formate dehydrogenase and method for producing the same
CN116676280A (en) Glutathione bifunctional synthetase mutant and application thereof
US20230332116A1 (en) Polypeptide with aspartate kinase activity and use thereof in production of amino acid
JP4162383B2 (en) Genes involved in the production of homoglutamic acid and use thereof
JPH11253162A (en) Dna encoding monoamine oxidase
US5804423A (en) Microbiological method of making 5-ketogluconate
JP4083455B2 (en) Aconitase gene of acetic acid bacteria, acetic acid bacteria bred using the gene, and method of producing vinegar using the acetic acid bacteria
US7811784B2 (en) Transgenic organisms with lower growth temperature
CN112062821B (en) Carbon catabolism regulatory protein CcpA mutant K31A
JP2938497B2 (en) Method for producing cytidine by fermentation
JPH05344891A (en) Gene capable of coding esterase and new microorganism containing the same gene
JPH0866189A (en) New dna fragment
JP2006109793A (en) METHOD FOR PRODUCING POLY-gamma-GLUTAMIC ACID AND MICROORGANISM USED IN THE PRODUCTION METHOD
JPH0731480A (en) Dna fragment coding l-glutamyl-trna reductase
CN116640751A (en) Ammonium aspartate ion lyase mutant and application thereof

Legal Events

Date Code Title Description
C10 Entry into substantive examination
SE01 Entry into force of request for substantive examination
C06 Publication
PB01 Publication
C10 Entry into substantive examination
SE01 Entry into force of request for substantive examination
C02 Deemed withdrawal of patent application after publication (patent law 2001)
WD01 Invention patent application deemed withdrawn after publication