CN102796779B - Biological method for preparing gamma-aminobutyric acid - Google Patents
Biological method for preparing gamma-aminobutyric acid Download PDFInfo
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- BTCSSZJGUNDROE-UHFFFAOYSA-N gamma-aminobutyric acid Chemical compound NCCCC(O)=O BTCSSZJGUNDROE-UHFFFAOYSA-N 0.000 title claims abstract description 183
- 229960003692 gamma aminobutyric acid Drugs 0.000 title claims abstract description 92
- OGNSCSPNOLGXSM-UHFFFAOYSA-N (+/-)-DABA Natural products NCCC(N)C(O)=O OGNSCSPNOLGXSM-UHFFFAOYSA-N 0.000 title claims abstract description 89
- 238000010170 biological method Methods 0.000 title abstract 3
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- 238000000034 method Methods 0.000 claims abstract description 28
- 238000002360 preparation method Methods 0.000 claims abstract description 18
- 241000894006 Bacteria Species 0.000 claims abstract description 16
- 239000013078 crystal Substances 0.000 claims abstract description 16
- 238000004519 manufacturing process Methods 0.000 claims abstract description 15
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- 238000000855 fermentation Methods 0.000 claims abstract description 12
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- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 claims abstract description 6
- 229910052799 carbon Inorganic materials 0.000 claims abstract description 6
- NWUYHJFMYQTDRP-UHFFFAOYSA-N 1,2-bis(ethenyl)benzene;1-ethenyl-2-ethylbenzene;styrene Chemical compound C=CC1=CC=CC=C1.CCC1=CC=CC=C1C=C.C=CC1=CC=CC=C1C=C NWUYHJFMYQTDRP-UHFFFAOYSA-N 0.000 claims description 16
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- 238000002425 crystallisation Methods 0.000 claims description 15
- 230000008025 crystallization Effects 0.000 claims description 15
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- FRXSZNDVFUDTIR-UHFFFAOYSA-N 6-methoxy-1,2,3,4-tetrahydroquinoline Chemical compound N1CCCC2=CC(OC)=CC=C21 FRXSZNDVFUDTIR-UHFFFAOYSA-N 0.000 claims description 13
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- 238000001035 drying Methods 0.000 claims description 10
- 239000003456 ion exchange resin Substances 0.000 claims description 10
- 229920003303 ion-exchange polymer Polymers 0.000 claims description 10
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- WHUUTDBJXJRKMK-UHFFFAOYSA-N Glutamic acid Natural products OC(=O)C(N)CCC(O)=O WHUUTDBJXJRKMK-UHFFFAOYSA-N 0.000 claims description 8
- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 claims description 7
- 239000000919 ceramic Substances 0.000 claims description 7
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- 235000013922 glutamic acid Nutrition 0.000 claims description 7
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- QWCKQJZIFLGMSD-UHFFFAOYSA-N 2-Aminobutanoic acid Natural products CCC(N)C(O)=O QWCKQJZIFLGMSD-UHFFFAOYSA-N 0.000 claims description 6
- QWCKQJZIFLGMSD-GSVOUGTGSA-N D-alpha-aminobutyric acid Chemical compound CC[C@@H](N)C(O)=O QWCKQJZIFLGMSD-GSVOUGTGSA-N 0.000 claims description 6
- 230000002378 acidificating effect Effects 0.000 claims description 6
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- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 claims description 5
- BNIILDVGGAEEIG-UHFFFAOYSA-L disodium hydrogen phosphate Chemical compound [Na+].[Na+].OP([O-])([O-])=O BNIILDVGGAEEIG-UHFFFAOYSA-L 0.000 claims description 5
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- SQQMAOCOWKFBNP-UHFFFAOYSA-L manganese(II) sulfate Chemical compound [Mn+2].[O-]S([O-])(=O)=O SQQMAOCOWKFBNP-UHFFFAOYSA-L 0.000 claims description 5
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- PXEDJBXQKAGXNJ-QTNFYWBSSA-L disodium L-glutamate Chemical compound [Na+].[Na+].[O-]C(=O)[C@@H](N)CCC([O-])=O PXEDJBXQKAGXNJ-QTNFYWBSSA-L 0.000 claims description 4
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- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 abstract description 16
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- 235000012538 ammonium bicarbonate Nutrition 0.000 description 1
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Abstract
The present invention discloses a biological method for preparing gamma-aminobutyric acid, including the steps of: preparing bacteria, producing gamma-aminobutyric acid, decoloring, conducting electrodialysis, preparing gamma-aminobutyric acid powder products or gamma-aminobutyric acid crystal products and the like. The present invention utilizes the free lactic acid bacterium cells for catalyzed synthesis of gamma-aminobutyric acid, and helps to improve the production technology, improve the gamma-aminobutyric acid yield, and reduce the cost, and also establishes the method for preparation of high-purity gamma-aminobutyric acid. The reaction system of the present method does not include carbon sources, nitrogen sources and other culture components, and the reaction liquid components are simple relative to fermentation liquid. The biological method can greatly improve the purity of the product, and has the advantages of simple post-treatment, short production cycle, little environmental pollution, low cost, etc.
Description
Technical field
The present invention relates to a kind of method for preparing γ-aminobutyric acid.
Background technology
γ-aminobutyric acid (Gamma-aminobutyric acid, GABA), claim again aminobutyric acid, is that a kind of naturally occurring nonprotein forms amino acid, is distributed widely in protokaryon and eukaryote.γ-aminobutyric acid is the inhibitory transmitter substance of mammalian central nervous system, have calm neural, promote sleep, reduce blood pressure, brain tonic and intelligence development, delay senility, the important physiological function such as the sharp kidney of strong liver.
Up to now, the preparation method of γ-aminobutyric acid mainly contains two kinds of chemical synthesis and biological processes.Common chemical synthesis process is as using pyrrolidone as raw material, through calcium hydroxide, bicarbonate of ammonia hydrolysis, makes γ-aminobutyric acid.The chemical synthesis cost is higher, and yield is lower, and uses dangerous solvents in production technique, or even deep-etching, noxious solvent.Therefore, the γ-aminobutyric acid of chemical synthesis preparation is difficult in the food factories realization, and goods can not be considered to a kind of natural additive for foodstuff.It is a kind of not only safety, cost but also low method that biological process is compared.According to up-to-date patent report, the bacterial classification that biological process is produced γ-aminobutyric acid has (1) natural original strain, comprises short lactobacillus (patent No. ZL 200510049187.5), aspergillus (publication number CN 101302480 B), plant lactobacillus (publication number CN 101928679 A), milk-acid bacteria (publication number CN 1243101 C) or natural food materials enrichment (publication number CN 1840672 A); (2) recombinant bacterium: intestinal bacteria (publication number CN 1298860 C), Corynebacterium glutamicum (publication number CN 101945997 A) (3) mutagenic strain: short lactobacillus (publication number CN 102174449 A).In existing method, the γ-aminobutyric acid fecund is born in fermented liquid, due to the fermented liquid complicated component, the downstream separation purge process is loaded down with trivial details, the concentration of target product is also lower, these effects limit domestic industry production, and about preparation high purity γ-aminobutyric acid, rarely have especially report.
Summary of the invention
The object of the present invention is to provide a kind of method easy, easy to operate, the method for effective Biological preparation γ-aminobutyric acid.
Technical solution of the present invention is:
A kind of method of Biological preparation γ-aminobutyric acid is characterized in that: comprise the following steps:
(1) thalline preparation:
The Sodium Glutamate that adds 1-30g/L in substratum, as inductor, is induced the product enzyme, and access is produced the bacterial strain of L-Glutamic decarboxylase in born of the same parents and cultivated; Culture condition is: 25～35 ℃ of culture temperature, carry out pH control by adding alkali lye in fermenting process, and fermented liquid pH is maintained in the 4.5-7.0 scope, incubation time 8～24 hours; Obtain bacterium liquid after cultivating end;
(2) ceramic membrane filter is collected thalline: the bacterium liquid that step (1) obtains is collected thalline through ceramic membrane filter, and membrane pore size is 0.05-1 μ m, obtains thalline;
(3) produce γ-aminobutyric acid:
The damping fluid of preparation 0.01-0.5mol/L pH3-7, add the 10-100g/L somatic cells, adds substrate glutamic acid sodium in batches, interpolation concentration is 0-30g/L, be under the condition of 28-40 ℃ in temperature of reaction, by fermentation again, produce γ-aminobutyric acid, fermentation time is 10-50h; After fermentation ends, by centrifugal collection clear liquid;
(4) clear liquid that step (3) is obtained is further removed somatic cells through metal micron membranes strainer, obtains clear liquid;
(5) decolouring: the clear liquid pH that first step (4) is obtained transfers to 7.0-8.0, then by the processing of decolour of granulated active carbon post, and the collection clear liquid;
(6) electrodialysis: adopt electrodialysis to remove impurity, controls material liquid pH is 7.0-8.0, drops in specific conductivity≤during 1000 μ s/cm, stop circulation, collection γ-aminobutyric acid clear liquid;
(7) the γ-aminobutyric acid clear liquid that step (6) is obtained, make γ-aminobutyric acid powder product or γ-aminobutyric acid crystal product.
The described method of making the γ-aminobutyric acid powder product comprises the following steps: successively
(a) vacuum concentration: adopt the triple effect plate-type evaporator to concentrate, when feed liquid is concentrated into alpha-aminobutyric acid content for 〉=80g/L, stop concentrating, collect concentrated solution;
(b) spraying drying: add maltodextrin or starch in concentrated solution, regulating solid quality content is 5-50%, carries out spraying drying, and making mass content is the γ-aminobutyric acid powder product of 20-50%.
The described method of making the γ-aminobutyric acid crystal product is: with the γ-aminobutyric acid clear liquid that step (6) obtains, and the process ion exchange resin treatment; Then material liquid pH is transferred to the γ-aminobutyric acid iso-electric point,, by decrease temperature crystalline, obtain the γ-aminobutyric acid crystal product.
In the described product of step (1) born of the same parents, the bacterial strain of L-Glutamic decarboxylase is milk-acid bacteria or lactobacillus hilgardii.
The composition of the substratum that step (1) is used and take the content of g/L as:
Glucose 10-30, yeast extract paste 5-30, peptone .5-30, sodium acetate 0.5-5, Sodium phosphate dibasic or dipotassium hydrogen phosphate 0.05-10, sal epsom 0.05-10, manganous sulfate 0.05-1, PPE polyether antifoam agent 0.1-3.
Described ion exchange resin is first selected anionite-exchange resin, to remove residual substrate glutamic acid sodium, then passes through acidic cation-exchange resin, to γ-aminobutyric acid adsorb, wash-out processes.
Described anionite-exchange resin is D301, D296 or 201 type highly basic or weak base anion-exchange resins; Described Zeo-karb is 001 or D113 type strong acid or weakly acidic cation-exchange resin.
The iso-electric point lowering temperature crystallization is adopted in crystallization, the feed liquid that obtains after ion exchange resin treatment is carried out crystallization or add ethanol and carry out crystallization, and adding proportion is ethanol: feed liquid=0～10:1V/V; During crystallization, regulator solution pH value is to the iso-electric point 7.2 of γ-aminobutyric acid, and the greenhouse cooling scope is 90～5 ℃, and the addition of crystal seed is 0. 2-10g/L, and stir speed (S.S.) is 30-100r/min, and rate of temperature fall is 1-10 ℃/h.
The present invention utilizes the free lactic acid mycetocyte to catalyze and synthesize γ-aminobutyric acid, and improvement of production process, and γ-aminobutyric acid output is improved, cost, but also set up the method for preparing the high purity γ-aminobutyric acid.Do not contain carbon source, nitrogenous source etc. in this technique in reaction system and cultivate composition, the reaction solution composition is simple for fermented liquid, can increase substantially product purity, has simultaneously that aftertreatment is simple, with short production cycle and environmental pollution is little, low cost and other advantages.
Description of drawings
The invention will be further described below in conjunction with drawings and Examples.
Fig. 1 is γ-aminobutyric acid formation curve in one embodiment of the invention preparation of industrialization process.
Fig. 2 transforms the amino acid detected result of the conversion fluid that generates γ-aminobutyric acid with lactic-acid bacteria cells.Glu: L-glutamic acid; GABA: γ-aminobutyric acid.
Embodiment
γ-aminobutyric acid of the present invention refers to 4-Aminobutanoicacid, is the mammalian central nervous system inhibitory transmitter., as new resource food, can add in beverage, cocoa products, chocolate and chocolate, candy, bakery product, puffed food the development of new product to.
Embodiment 1:
One, yeast culture
After lactic acid bacteria culturers is activated, carrying out first order seed cultivates, secondary seed is cultivated, enter finally fermentor cultivation, inoculum size 3%(V/V, seed liquor/fermented liquid), 30 ℃ of standing cultivation 16 h, carry out pH control by adding alkali lye in fermenting process, fermented liquid pH is maintained in the 4.5-7.0 scope, by 0.6 μ m ceramic membrane filter, collect thalline.
The seed culture based raw material is in g/L: glucose 10, yeast extract paste 5, peptone 5, sodium acetate 0.5, Sodium phosphate dibasic (or dipotassium hydrogen phosphate) 1, sal epsom 1, manganous sulfate 1, PPE polyether antifoam agent 0.1(but be not limited only to this defoamer, can select any defoamer that can be used for fermentation industry).
Fermentative medium formula is in g/L: glucose 10, yeast extract paste 5, peptone 5, sodium acetate 0.5, Sodium Glutamate 1, Sodium phosphate dibasic (or dipotassium hydrogen phosphate) 1, sal epsom 1, manganous sulfate 1, PPE polyether antifoam agent 0.1(but be not limited only to this defoamer, can select any defoamer that can be used for fermentation industry).
Two, produce γ-aminobutyric acid
Adopt the lactic-acid bacteria cells conversion method to produce γ-aminobutyric acid, reaction conditions is: acetic acid-sodium-acetate buffer of preparation 0.01mol/L pH7, add the 20g/L somatic cells, add substrate glutamic acid sodium in batches, initial substrate concentration 10g/L, every 4h adds a substrate, interpolation concentration is 5g/L, the total reaction concentration of substrate is 25g/L, and temperature of reaction is 28 ℃, and fermentation time is 17h.After end,, by centrifugal collection clear liquid, obtain content and reach the γ-aminobutyric acid conversion fluid of 12g/L, see Fig. 1.
Three, the detection of alpha-aminobutyric acid content
The trichoroacetic acid(TCA) that adds equal-volume 100g/L after above-mentioned conversion fluid is centrifugal, vibration is even, more centrifugal (10,000 * g, 5 min), 2-10 times of clear liquid dilution, then, through 0.45 μ m membrane filtration, carry out high-efficient liquid phase analysis.Adopt ODS C
18(φ 4.6 * 250nm), and 40
oC analyzes, and γ-aminobutyric acid is quantitative with external standard method.
Four, product preparation:
Filtration sterilization: the γ-aminobutyric acid conversion fluid that will obtain is further removed somatic cells through metal micron membranes strainer, obtains clear liquid;
Decolouring: the clear liquid pH that obtains after filtration sterilization is transferred to 7.0-8.0, then by the processing of decolour of granulated active carbon post, the collection clear liquid;
Electrodialysis: adopt electrodialysis to remove impurity, controls material liquid pH is 7.0-8.0, drops in specific conductivity≤during 1000 μ s/cm, stop circulation, collection γ-aminobutyric acid clear liquid;
, with the γ-aminobutyric acid clear liquid that step (4) obtains, make γ-aminobutyric acid powder product or γ-aminobutyric acid crystal product.
The described method of making the γ-aminobutyric acid powder product comprises the following steps: successively
(a) vacuum concentration: adopt the triple effect plate-type evaporator to concentrate, when feed liquid is concentrated into alpha-aminobutyric acid content for 〉=50g/L, stop concentrating;
(b) spraying drying: add maltodextrin or starch in concentrated solution, regulating solid quality content is 5-50%, carries out spraying drying, and making mass content is the γ-aminobutyric acid powder product of 20-50%;
The described method of making the γ-aminobutyric acid powder product comprises the following steps: successively
(a) vacuum concentration: adopt the triple effect plate-type evaporator to concentrate, when feed liquid is concentrated into alpha-aminobutyric acid content for 〉=80g/L, stop concentrating, collect concentrated solution;
(b) spraying drying: add maltodextrin or starch in concentrated solution, regulating solid quality content is 5-50%, carries out spraying drying, and making mass content is the γ-aminobutyric acid powder product of 20-50%.
The described method of making the γ-aminobutyric acid crystal product is: with the γ-aminobutyric acid clear liquid that step (6) obtains, and the process ion exchange resin treatment; Then material liquid pH is transferred to the γ-aminobutyric acid iso-electric point,, by decrease temperature crystalline, obtain purity and reach 98% γ-aminobutyric acid crystal product.
Embodiment 2:
A kind of method of Biological preparation γ-aminobutyric acid, comprise the following steps:
(1) thalline preparation:
Add Sodium Glutamate in substratum, addition is 1-30g/L(example 1 g/L, 15 g/L, 30 g/L) as inductor, induce the product enzyme, access is produced the bacterial strain of L-Glutamic decarboxylase in born of the same parents and is cultivated; Culture condition is: 25～35 ℃ of culture temperature (25 ℃, 30 ℃, 35 ℃ of examples), carry out pH control by adding alkali lye in fermenting process, and fermented liquid pH is maintained in the 4.5-7.0 scope, incubation time 8～24 hours (routine 8h, 16h, 24h); Obtain bacterium liquid after cultivating end;
(2) ceramic membrane filter is collected thalline: the bacterium liquid that step (1) obtains is collected thalline through ceramic membrane filter, and membrane pore size is 0.05-1 μ m(example 0.05 μ m, 0.5 μ m, 1 μ m), obtain thalline;
(3) produce γ-aminobutyric acid:
Preparation 0.01-0.5mol/L(example 0.01 mol/L, 0.2 mol/L, 0.5 mol/L) pH3-7(example 3,5,7) damping fluid, add somatic cells, the somatic cells add-on is 10-100g/L(example 10 g/L, 50 g/L, 100 g/L), add substrate glutamic acid sodium in batches, adding concentration is 1-30g/L(example 1g/L, 15 g/L, 30 g/L), be under the condition of 28-40 ℃ (28 ℃, 32 ℃, 40 ℃ of examples) in temperature of reaction, by again fermenting and produce γ-aminobutyric acid, fermentation time is 10-50h(example 10h, 25h, 50h); After fermentation ends, by centrifugal collection clear liquid;
(4) clear liquid that step (3) is obtained is further removed somatic cells through metal micron membranes strainer, obtains clear liquid;
(5) decolouring: the clear liquid pH that first step (4) is obtained transfers to 7.0-8.0, then by the processing of decolour of granulated active carbon post, and the collection clear liquid;
(6) electrodialysis: adopt electrodialysis to remove impurity, controls material liquid pH is 7.0-8.0, drops in specific conductivity≤during 1000 μ s/cm, stop circulation, collection γ-aminobutyric acid clear liquid;
(7) the γ-aminobutyric acid clear liquid that step (6) is obtained, make γ-aminobutyric acid powder product or γ-aminobutyric acid crystal product.
The described method of making the γ-aminobutyric acid powder product comprises the following steps: successively
(a) vacuum concentration: adopt the triple effect plate-type evaporator to concentrate, when feed liquid is concentrated into alpha-aminobutyric acid content for 〉=80g/L, stop concentrating, collect concentrated solution;
(b) spraying drying: add maltodextrin or starch in concentrated solution, regulating solid quality content is 5-50%(example 5%, 30%, 50%), carry out spraying drying, making mass content is the γ-aminobutyric acid powder product of 20-50%.
The described method of making the γ-aminobutyric acid crystal product is: with the γ-aminobutyric acid clear liquid that step (6) obtains, and the process ion exchange resin treatment; Then material liquid pH is transferred to the γ-aminobutyric acid iso-electric point,, by decrease temperature crystalline, obtain the γ-aminobutyric acid crystal product.
In the described product of step (1) born of the same parents, the bacterial strain of L-Glutamic decarboxylase is milk-acid bacteria or lactobacillus hilgardii.
The composition of the substratum that step (1) is used and take the content of g/L as:
Glucose 10-30(example 10,20,30), yeast extract paste 5-30(example 5,20,30), peptone .5-30(example 5,20,30), sodium acetate 0.5-5(example 0.5,3,5), Sodium phosphate dibasic or dipotassium hydrogen phosphate 0.05-10(example 0.05,5,10), sal epsom 0.05-10(example 0.05,5,10), manganous sulfate 0.05-1(example 0.05,0.5,1), PPE polyether antifoam agent 0.1-3(example 0.1,1,3).
Described ion exchange resin is first selected anionite-exchange resin, to remove residual substrate glutamic acid sodium, then passes through acidic cation-exchange resin, to γ-aminobutyric acid adsorb, wash-out processes.
Described anionite-exchange resin is D301, D296 or 201 type highly basic or weak base anion-exchange resins; Described Zeo-karb is 001 or D113 type strong acid or weakly acidic cation-exchange resin.
The iso-electric point lowering temperature crystallization is adopted in crystallization, the feed liquid that obtains after ion exchange resin treatment is carried out crystallization or add ethanol and carry out crystallization, and adding proportion is ethanol: feed liquid=0～10:1V/V(example 1:1,5:1,10:1); During crystallization, regulator solution pH value is to the iso-electric point 7.2 of γ-aminobutyric acid, the greenhouse cooling scope is 90～5 ℃ (90 ℃, 40 ℃, 5 ℃ of examples), the addition of crystal seed is 0. 2-10g/L(example 0.2 g/L, 5 g/L, 10 g/L), stir speed (S.S.) is 30-100r/min, and rate of temperature fall is 1 ℃/h of 1-10 ℃/h(example, 5 ℃/h, 10 ℃/h).
Claims (1)
1. the method for a Biological preparation γ-aminobutyric acid, is characterized in that: comprise the following steps:
(1) thalline preparation:
The Sodium Glutamate that adds 1-30g/L in substratum, as inductor, is induced the product enzyme, and access is produced the bacterial strain of L-Glutamic decarboxylase in born of the same parents and cultivated; Culture condition is: 25～35 ℃ of culture temperature, carry out pH control by adding alkali lye in fermenting process, and fermented liquid pH is maintained in the 4.5-7.0 scope, incubation time 8～24 hours; Obtain bacterium liquid after cultivating end; The composition of the substratum that uses and take the content of g/L as:
Glucose 10-30, yeast extract paste 5-30, peptone 5-30, sodium acetate 0.5-5, Sodium phosphate dibasic or dipotassium hydrogen phosphate 0.05-10, sal epsom 0.05-10, manganous sulfate 0.05-1, PPE polyether antifoam agent 0.1-3;
(2) ceramic membrane filter is collected thalline: the bacterium liquid that step (1) obtains is collected thalline through ceramic membrane filter, and membrane pore size is 0.05-1 μ m, obtains thalline;
(3) produce γ-aminobutyric acid:
The damping fluid of preparation 0.01-0.5mol/L pH3-7, add the 10-100g/L somatic cells, adds substrate glutamic acid sodium in batches, interpolation concentration is 25g/L, be under the condition of 28-40 ℃ in temperature of reaction, by fermentation again, produce γ-aminobutyric acid, fermentation time is 10-50h; After fermentation ends, by centrifugal collection clear liquid;
(4) clear liquid that step (3) is obtained is further removed somatic cells through metal micron membranes strainer, obtains clear liquid;
(5) decolouring: the clear liquid pH that first step (4) is obtained transfers to 7.0-8.0, then by the processing of decolour of granulated active carbon post, and the collection clear liquid;
(6) electrodialysis: adopt electrodialysis to remove impurity, controls material liquid pH is 7.0-8.0, drops in specific conductivity≤during 1000 μ s/cm, stop circulation, collection γ-aminobutyric acid clear liquid;
(7) the γ-aminobutyric acid clear liquid that step (6) is obtained, make γ-aminobutyric acid powder product or γ-aminobutyric acid crystal product;
The described method of making the γ-aminobutyric acid powder product comprises the following steps: successively
(a) vacuum concentration: adopt the triple effect plate-type evaporator to concentrate, when feed liquid is concentrated into alpha-aminobutyric acid content for 〉=80g/L, stop concentrating, collect concentrated solution;
(b) spraying drying: add maltodextrin or starch in concentrated solution, regulating solid quality content is 5-50%, carries out spraying drying, and making mass content is the γ-aminobutyric acid powder product of 20-50%;
The described method of making the γ-aminobutyric acid crystal product is: with the γ-aminobutyric acid clear liquid that step (6) obtains, and the process ion exchange resin treatment; Then material liquid pH is transferred to the γ-aminobutyric acid iso-electric point,, by decrease temperature crystalline, obtain the γ-aminobutyric acid crystal product; Described ion exchange resin is first selected anionite-exchange resin, to remove residual substrate glutamic acid sodium, then passes through acidic cation-exchange resin, to γ-aminobutyric acid adsorb, wash-out processes; Described anionite-exchange resin is D301, D296 or 201 type highly basic or weak base anion-exchange resins; Described Zeo-karb is 001 or D113 type strong acid or weakly acidic cation-exchange resin; The iso-electric point lowering temperature crystallization is adopted in crystallization, the feed liquid that obtains after ion exchange resin treatment is carried out crystallization or add ethanol and carry out crystallization, and adding proportion is ethanol: feed liquid=1～10:1(V/V); During crystallization, regulator solution pH value is to the iso-electric point 7.2 of γ-aminobutyric acid, and the greenhouse cooling scope is 90～5 ℃, and the addition of crystal seed is 0. 2-10g/L, and stir speed (S.S.) is 30-100r/min, and rate of temperature fall is 1-10 ℃/h.
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| CN102978250B (en) * | 2012-12-20 | 2014-07-30 | 江苏久吾高科技股份有限公司 | Method for producing Gamma-aminobutyric acid through centrifugal mother liquid of glutamic acid |
| CN104561160A (en) * | 2014-12-22 | 2015-04-29 | 南通励成生物工程有限公司 | Method for preparing theanine by using biological method |
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| CN101333548A (en) * | 2008-06-26 | 2008-12-31 | 南昌大学 | Method for preparing gamma-aminobutyric acid by lactobacillus brevis |
| CN101838672B (en) * | 2010-05-10 | 2014-12-17 | 江南大学 | Method for producing gamma-amino butyric acid by using immobilized lactobacillus plantarum |
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