CN103695485B - High-efficiency production method of gamma-polyglutamic acid - Google Patents
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- 238000004519 manufacturing process Methods 0.000 title abstract description 14
- 229920002643 polyglutamic acid Polymers 0.000 title abstract description 8
- 238000000855 fermentation Methods 0.000 claims abstract description 63
- 230000004151 fermentation Effects 0.000 claims abstract description 63
- 241000194108 Bacillus licheniformis Species 0.000 claims abstract description 14
- 238000000034 method Methods 0.000 claims abstract description 14
- 238000009423 ventilation Methods 0.000 claims abstract description 13
- VWDWKYIASSYTQR-UHFFFAOYSA-N sodium nitrate Chemical compound [Na+].[O-][N+]([O-])=O VWDWKYIASSYTQR-UHFFFAOYSA-N 0.000 claims abstract description 11
- 239000001963 growth medium Substances 0.000 claims abstract description 6
- 239000002609 medium Substances 0.000 claims description 38
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 claims description 22
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 18
- 239000007788 liquid Substances 0.000 claims description 15
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims description 13
- 239000008103 glucose Substances 0.000 claims description 13
- 229910052943 magnesium sulfate Inorganic materials 0.000 claims description 11
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 claims description 10
- 239000001110 calcium chloride Substances 0.000 claims description 10
- 229910001628 calcium chloride Inorganic materials 0.000 claims description 10
- SQQMAOCOWKFBNP-UHFFFAOYSA-L manganese(II) sulfate Chemical compound [Mn+2].[O-]S([O-])(=O)=O SQQMAOCOWKFBNP-UHFFFAOYSA-L 0.000 claims description 10
- 229910000357 manganese(II) sulfate Inorganic materials 0.000 claims description 10
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 8
- 229940041514 candida albicans extract Drugs 0.000 claims description 8
- 239000012138 yeast extract Substances 0.000 claims description 8
- 238000002360 preparation method Methods 0.000 claims description 7
- 230000004913 activation Effects 0.000 claims description 6
- 230000001133 acceleration Effects 0.000 claims description 5
- 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 5
- 239000002054 inoculum Substances 0.000 claims description 5
- 229910000359 iron(II) sulfate Inorganic materials 0.000 claims description 5
- 235000013923 monosodium glutamate Nutrition 0.000 claims description 5
- 229940073490 sodium glutamate Drugs 0.000 claims description 5
- 239000012530 fluid Substances 0.000 claims description 4
- 239000011780 sodium chloride Substances 0.000 claims description 4
- 229920001817 Agar Polymers 0.000 claims description 3
- 239000001888 Peptone Substances 0.000 claims description 3
- 108010080698 Peptones Proteins 0.000 claims description 3
- 240000004808 Saccharomyces cerevisiae Species 0.000 claims description 3
- 239000008272 agar Substances 0.000 claims description 3
- 229910000396 dipotassium phosphate Inorganic materials 0.000 claims description 3
- 235000019319 peptone Nutrition 0.000 claims description 3
- 239000000843 powder Substances 0.000 claims description 3
- 239000012137 tryptone Substances 0.000 claims description 3
- 230000009123 feedback regulation Effects 0.000 claims description 2
- 229910052564 epsomite Inorganic materials 0.000 claims 1
- 229910052930 hexahydrite Inorganic materials 0.000 claims 1
- WHUUTDBJXJRKMK-UHFFFAOYSA-N Glutamic acid Natural products OC(=O)C(N)CCC(O)=O WHUUTDBJXJRKMK-UHFFFAOYSA-N 0.000 abstract description 12
- 235000013922 glutamic acid Nutrition 0.000 abstract description 12
- 239000004220 glutamic acid Substances 0.000 abstract description 12
- 229910002651 NO3 Inorganic materials 0.000 abstract description 5
- 238000006243 chemical reaction Methods 0.000 abstract description 4
- 230000000694 effects Effects 0.000 abstract description 3
- 239000002861 polymer material Substances 0.000 abstract description 3
- 235000010344 sodium nitrate Nutrition 0.000 abstract description 3
- 239000004317 sodium nitrate Substances 0.000 abstract description 3
- NHNBFGGVMKEFGY-UHFFFAOYSA-N Nitrate Chemical compound [O-][N+]([O-])=O NHNBFGGVMKEFGY-UHFFFAOYSA-N 0.000 abstract description 2
- 230000029058 respiratory gaseous exchange Effects 0.000 abstract description 2
- 101000681023 Escherichia coli (strain K12) Ribosomal protein S6-L-glutamate ligase Proteins 0.000 abstract 1
- 238000009629 microbiological culture Methods 0.000 abstract 1
- WHUUTDBJXJRKMK-VKHMYHEASA-N L-glutamic acid Chemical compound OC(=O)[C@@H](N)CCC(O)=O WHUUTDBJXJRKMK-VKHMYHEASA-N 0.000 description 25
- 229960002989 glutamic acid Drugs 0.000 description 17
- 230000001580 bacterial effect Effects 0.000 description 6
- 239000000463 material Substances 0.000 description 4
- 238000001514 detection method Methods 0.000 description 3
- QGZKDVFQNNGYKY-UHFFFAOYSA-N Ammonia Chemical compound N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 description 2
- 108010020346 Polyglutamic Acid Proteins 0.000 description 2
- 239000002253 acid Substances 0.000 description 2
- 230000001419 dependent effect Effects 0.000 description 2
- 239000003814 drug Substances 0.000 description 2
- 244000005700 microbiome Species 0.000 description 2
- 239000002243 precursor Substances 0.000 description 2
- 238000011218 seed culture Methods 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- DWNBOPVKNPVNQG-LURJTMIESA-N (2s)-4-hydroxy-2-(propylamino)butanoic acid Chemical compound CCCN[C@H](C(O)=O)CCO DWNBOPVKNPVNQG-LURJTMIESA-N 0.000 description 1
- FRXSZNDVFUDTIR-UHFFFAOYSA-N 6-methoxy-1,2,3,4-tetrahydroquinoline Chemical compound N1CCCC2=CC(OC)=CC=C21 FRXSZNDVFUDTIR-UHFFFAOYSA-N 0.000 description 1
- 241000193830 Bacillus <bacterium> Species 0.000 description 1
- 241000311115 Bacillus paralicheniformis ATCC 9945a Species 0.000 description 1
- 244000063299 Bacillus subtilis Species 0.000 description 1
- 235000014469 Bacillus subtilis Nutrition 0.000 description 1
- 241000894006 Bacteria Species 0.000 description 1
- WHUUTDBJXJRKMK-GSVOUGTGSA-N D-glutamic acid Chemical compound OC(=O)[C@H](N)CCC(O)=O WHUUTDBJXJRKMK-GSVOUGTGSA-N 0.000 description 1
- 229930182847 D-glutamic acid Natural products 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 241001597008 Nomeidae Species 0.000 description 1
- 239000002250 absorbent Substances 0.000 description 1
- 230000002745 absorbent Effects 0.000 description 1
- 125000003368 amide group Chemical group 0.000 description 1
- 229910021529 ammonia Inorganic materials 0.000 description 1
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- 229910001385 heavy metal Inorganic materials 0.000 description 1
- 238000009655 industrial fermentation Methods 0.000 description 1
- 238000011031 large-scale manufacturing process Methods 0.000 description 1
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- 231100000252 nontoxic Toxicity 0.000 description 1
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- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention discloses a high-efficiency production method of gamma-polyglutamic acid, and belongs to the technical field of production of polymer materials based on biological fermentation. According to the method, bacillus licheniformis CGMCC (China General Microbiological Culture Collection Center) NO.3336 for high-efficient production of gamma-polyglutamic acid is taken as a starting strain, sodium nitrate is added into a fermentation culture medium, and the activity of polyglutamate synthase is influenced under the nitrate respiration condition by adjusting and controlling the pH value and the ventilation quantity in a fermentation process, so that residual glutamic acid in a fermentation solution is reduced and the conversion rate of glutamic acid is increased; after fermentation, the glutamic acid content of the fermentation solution is 5-15 g/L and the gamma-polyglutamic acid yield is 45-55 g/L. Therefore, the economic and feasible production method is provided for industrialization production of gamma-polyglutamic acid.
Description
Technical field
The invention belongs to biofermentation produces technical field of polymer materials, specifically a kind of efficiently production γ-polyglutamic
The method of acid.
Background technology
Gamma-polyglutamic acid-is by the amido link combination between alpha-amido and γ-carboxyl by D-Glu and L-Glutamic Acid
A kind of multifunctional bio degradable high polymer material becoming.Because gamma-polyglutamic acid-and its derivant have excellent water solublity
And adsorptivity, can thoroughly be biodegradable, to humans and animals safety non-toxic, the advantages of edible, thus can as water-retaining agent,
Thickening agent, flocculant, heavy metal absorbent, medicine, fertilizer slow-release formulation and pharmaceutical carrier etc., in agricultural, food, medicine, cosmetic
The fields such as product, environmental protection, synthetic fibers and film have broad application prospects.The environmental pollution causing with chemical industry synthesis material
Increasingly serious, exploitation Biodegradable material substitutes chemical material to a certain extent and has become the urgent of whole international community and is essential
Ask.
The preparation method of polyglutamic acid has chemosynthesis, extraction and three kinds of methods of fermentable.Microbe fermentation method ratio
The low production cost of first two method, production process is little to the pollution of environment, so primarily now adopting microbe fermentation method to give birth to
Produce gamma-polyglutamic acid-.
Both at home and abroad the fermentation technology of gamma-polyglutamic acid-is conducted extensive research, mainly with the training of liquid submerged fermentation
Based on foster based formulas and culture process.One plant of gamma-polyglutamic acid-superior strain Bcillius screens to obtain in Nanjing University of Technology
Subtilis NX-2, produces gamma-polyglutamic acid-and has done the more comprehensive research of ratio, and applied for patent, patent is public to liquid fermentation
The number of opening is CN1346891.Hua Zhong Agriculture University has been also carried out the screening of gamma-polyglutamic acid generating bacterium and the work of process exploitation,
Its strain and liquids production process have also applied for patent, and patent publication No. is CN1536071.Korea S's research worker is 2.5
Rise the method in fermentation tank, using stream, High Density Cultivation being added to Bacillus licheniformis ATCC9945a, fermentation 35 is little
When after can reach the whole yield of 35g/L（Yoon SH, DO JH, Lee SY.Biotechnol.Lett, 2000,22:585-
588）.Kubota separates, in Osaka City University soil, the one plant of Bacillus subtilis F201 obtaining,
This bacterial strain can reach the maximum output of 50g/L under optimal fermentation conditions, and this is the maximum output of document report（Kubota
H.Biosci Biotec Biochem, 1993:1212-1213）.
γ-the PGA having now been found that produces bacterial strain and is mainly bacillus.According to whether providing L- paddy ammonia in culture medium
Bacterial strain can be divided into two big class by acid：One class is the L-Glutamic Acid dependent form needing to add in the medium L-Glutamic Acid precursor,
Another kind of for not needing to add in the medium the L-Glutamic Acid independent form of L-Glutamic Acid precursor.L-Glutamic Acid independent form is given birth to
Produce less efficient, so industrial fermentation produces substantially all selects L-Glutamic Acid dependent form bacterial strain, L-Glutamic Acid is as before important
Body is added in the medium, but its 50% about conversion ratio is not appropriate for economical large-scale production, seeks a kind of height
Effect, the gamma-polyglutamic acid-production method of low cost are still a kind of needs.
Content of the invention
Technical problem solved by the invention is with the Bacillus licheniformis of high yield gamma-polyglutamic acid-（Bacillus
licheniformis）CGMCC NO.3336 is starting strain, adds sodium nitrate and fermented by regulation and control in fermentation medium
During pH value and ventilation to reduce the residue of fermentation liquid Glutamic Acid, improve the conversion ratio of glutamic acid, with reduces cost, carry
The yield of high gamma-polyglutamic acid-.
The bacterial strain of the production high molecular gamma-polyglutamic acid-that the present invention provides is specially Bacillus licheniformis（Bacillus
licheniformis）CGMCC NO.3336.This bacterial strain is preserved in Chinese microorganism strain preservation on October 14th, 2009
Administration committee's common micro-organisms center（Abbreviation CGMCC, address：Datun Road, Chaoyang District, Beijing City, Chinese Academy of Sciences's microorganism is ground
Study carefully institute, postcode：100101）, preserving number is CGMCC NO.3336, and Classification And Nomenclature is：Bacillus licheniformis（Bacillus
licheniformis）.
A kind of efficient method producing gamma-polyglutamic acid-, comprises the steps：
（1）Actication of culture：Bacillus licheniformis CGMCC NO.3336 original strain is seeded on slant medium, in
Cultivate 16 hours for 37 DEG C, so activation 2-3 time, the ripe inclined-plane seed of preparation；
Described slant medium consists of：Peptone 10g/L, yeast powder 5g/L, NaCl10g/L, agar 20g/L,
PH7.2, insufficient section pure water is supplied；
（2）The preparation of seed liquor：Inclined-plane seed after one ring activation is inoculated in the 500ml equipped with 50ml fluid medium
In triangular flask, 37 DEG C, 220rpm cultivates 16 hours to exponential phase；
Described seed culture medium consists of：Glucose 30g/L, yeast extract 7g/L, tryptone 10g/L, K2HPO40.5g/
L, MgSO4·7H2O0.5g/L, pH7.2, insufficient section pure water is supplied；
（3）Ferment tank：By step（2）In seed liquor fermentation tank, 30L fermentation tank are inoculated in 10% inoculum concentration
Fermentation medium liquid amount is 15L, 37 DEG C of Preliminary fermentation temperature, tank pressure 0.01-0.05Mpa, ventilation 0.5-1.0vvm, rotating speed
400-600rpm；Adjust rotating speed to 200-400rpm, ventilation 1.0-1.5vvm after fermentation is to 24h, and start to add stream plus
Culture medium, by the flow acceleration of the feedback regulation fed-batch medium of pH value, control ph 2- before 7.4-7.9 to fermentation ends
6 hours, total fermentation time was 72h.
Described fermentation medium consists of：Glucose 80g/L, yeast extract 20g/L, NH4NO34.5g/L, MgSO4·
6H2O0.5g/L, NaCl10g/L, CaCl2·6H2O0.5g/L, FeSO40.01g/L, sodium glutamate 40g/L, NaNO320-40g/
L,MnSO4·H2O0.15g/L, pH value 7.2-7.5, insufficient section pure water is supplied；
Described fed-batch medium consists of：Glucose 650-850g/L, NaCl30.0-60.0g/L, CaCl2·6H2O10-
15.0g/L, MnSO4·H2O5-10g/L, MgSO4·6H2O5-20g/L, pH value 7.4-7.9, insufficient section pure water is supplied；
After fermentation ends, thalline is removed in centrifugation, takes 100 times of supernatant distilled water diluting, by SBA-40E bio-sensing instrument
Remaining glutamic acid is detected, content of glutamic acid is 5-15g/L, gamma-polyglutamic acid-yield 45-55g/L.
Beneficial effect：
The present invention is with the Bacillus licheniformis of high yield gamma-polyglutamic acid-（Bacillus licheniformis）CGMCC
NO.3336 is starting strain, adds sodium nitrate and by regulating and controlling pH value and ventilation in sweat in fermentation medium,
Affecting the activity of polyglutamic acid synzyme under the conditions of nitrate respiration, thus reducing the residue of fermentation liquid Glutamic Acid, improving paddy
The conversion ratio of propylhomoserin, fermentation ends after fermentation liquid Glutamic Acid content is 5-15g/L, gamma-polyglutamic acid-yield 45-55g/L.For
Gamma-polyglutamic acid-industrialized production provides a kind of economically viable production method.
Specific embodiment
Describe the present invention below by specific embodiment.Unless stated otherwise, technological means used in the present invention
It is method known in those skilled in the art.In addition, embodiment is interpreted as illustrative, and the unrestricted present invention
Scope, the spirit and scope of the invention are limited only by the claims that follow.To those skilled in the art, without departing substantially from this
On the premise of invention spirit and scope, the material component in these embodiments and consumption are carried out various changes or are changed
Belong to protection scope of the present invention.
Embodiment 1
A kind of efficient method producing gamma-polyglutamic acid-, comprises the steps：
（1）Actication of culture：Bacillus licheniformis CGMCC NO.3336 original strain is seeded on slant medium, in
Cultivate 16 hours for 37 DEG C, so activation 2 times, the ripe inclined-plane seed of preparation；
Described slant medium consists of：Peptone 10g/L, yeast powder 5g/L, NaCl10g/L, agar 20g/L,
PH7.2, insufficient section pure water is supplied；
（2）The preparation of seed liquor：Inclined-plane seed after one ring activation is inoculated in the 500ml equipped with 50ml fluid medium
In triangular flask, 37 DEG C, 220rpm cultivates 16 hours to exponential phase；
Described seed culture medium consists of：Glucose 30g/L, yeast extract 7g/L, tryptone 10g/L, K2HPO40.5g/
L, MgSO4·7H2O0.5g/L, pH7.2, insufficient section pure water is supplied；
（3）Ferment tank：By step（2）In seed liquor fermentation tank, 30L fermentation tank are inoculated in 10% inoculum concentration
Fermentation medium liquid amount is 15L, 37 DEG C of Preliminary fermentation temperature, tank pressure 0.03Mpa ventilation 0.8vvm, rotating speed 500rpm；When
Ferment to 24h and adjust rotating speed to 300rpm, ventilation 1.2vvm, and start to add fed-batch medium, by the feedback of pH value
Adjust the flow acceleration of fed-batch medium, control ph first 4 hours in 7.7 to fermentation ends, total fermentation time is 72h.
Described fermentation medium consists of：Glucose 80g/L, yeast extract 20g/L, NH4NO34.5g/L, MgSO4·
6H2O0.5g/L, NaCl10g/L, CaCl2·6H2O0.5g/L, FeSO40.01g/L, sodium glutamate 40g/L, NaNO330g/L,
MnSO4·H2O0.15g/L, pH value 7.2, insufficient section pure water is supplied；
Described fed-batch medium consists of：Glucose 750g/L, NaCl40g/L, CaCl2·6H2O12g/L, MnSO4·
H2O8g/L, MgSO4·6H2O12g/L, pH value 7.7, insufficient section pure water is supplied；
Through detection after fermentation ends, fermentation liquid Glutamic Acid content is 10g/L, gamma-polyglutamic acid-yield 50g/L.
Embodiment 2
Actication of culture and seed liquor are prepared with embodiment 1
（3）Ferment tank：By step（2）In seed liquor fermentation tank, 30L fermentation tank are inoculated in 10% inoculum concentration
Fermentation medium liquid amount is 15L, 37 DEG C of Preliminary fermentation temperature, tank pressure 0.01Mpa ventilation 0.5vvm, rotating speed 400rpm；When
Ferment to 24h and adjust rotating speed to 200rpm, ventilation 1.0vvm, and start to add fed-batch medium, by the feedback of pH value
Adjust the flow acceleration of fed-batch medium, control ph first 2 hours in 7.4 to fermentation ends, total fermentation time is 72h.
Described fermentation medium consists of：Glucose 80g/L, yeast extract 20g/L, NH4NO34.5g/L, MgSO4·
6H2O0.5g/L, NaCl10g/L, CaCl2·6H2O0.5g/L, FeSO40.01g/L, sodium glutamate 40g/L, NaNO320g/L,
MnSO4·H2O0.15g/L, pH value 7.4, insufficient section pure water is supplied；
Described fed-batch medium consists of：Glucose 650g/L, NaCl30.0g/L, CaCl2·6H2O10g/L, MnSO4·
H2O5g/L, MgSO4·6H2O5g/L, pH value 7.4, insufficient section pure water is supplied；
Through detection after fermentation ends, fermentation liquid Glutamic Acid content is 15g/L, gamma-polyglutamic acid-yield 45g/L.
Embodiment 3
（3）Ferment tank：By step（2）In seed liquor fermentation tank, 30L fermentation tank are inoculated in 10% inoculum concentration
Fermentation medium liquid amount is 15L, 37 DEG C of Preliminary fermentation temperature, tank pressure 0.05Mpa ventilation 1.0vvm, rotating speed 600rpm；When
Ferment to 24h and adjust rotating speed to 400rpm, ventilation 1.5vvm, and start to add fed-batch medium, by the feedback of pH value
Adjust the flow acceleration of fed-batch medium, control ph first 6 hours in 7.9 to fermentation ends, total fermentation time is 72h.
Described fermentation medium consists of：Glucose 80g/L, yeast extract 20g/L, NH4NO34.5g/L, MgSO4·
6H2O0.5g/L, NaCl10g/L, CaCl2·6H2O0.5g/L, FeSO40.01g/L, sodium glutamate 40g/L, NaNO340g/L,
MnSO4·H2O0.15g/L, pH value 7.5, insufficient section pure water is supplied；
Described fed-batch medium consists of：Glucose 850g/L, sodium chloride 60.0g/L, CaCl2·6H2O15.0g/L,
MnSO4·H2O10g/L, MgSO4·6H2O20g/L, pH value 7.9, insufficient section pure water is supplied；
Through detection after fermentation ends, fermentation liquid Glutamic Acid content is 5g/L, gamma-polyglutamic acid-yield 55g/L.
Claims (1)
1. a kind of efficient produce gamma-polyglutamic acid-method it is characterised in that：With Bacillus licheniformis (Bacillus
Licheniformis) CGMCC NO.3336 is starting strain, is prepared by liquid submerged fermentation, comprises the steps：
(1) actication of culture：Bacillus licheniformis CGMCC NO.3336 original strain is seeded on slant medium, in 37 DEG C
Culture 16 hours, so activation 2-3 time, the ripe inclined-plane seed of preparation；
Described slant medium consists of：Peptone 10g/L, yeast powder 5g/L, NaCl 10g/L, agar 20g/L, pH7.2, no
Foot divides pure water to supply；
(2) preparation of seed liquor：Inclined-plane seed after one ring activation is inoculated in the 500ml triangle equipped with 50ml fluid medium
In bottle, 37 DEG C, 220rpm cultivates 16 hours to exponential phase；
Described fluid medium consists of：Glucose 30g/L, yeast extract 7g/L, tryptone 10g/L, K2HPO40.5g/L,
MgSO4·7H2O 0.5g/L, pH7.2, insufficient section pure water is supplied；
(3) ferment tank：Seed liquor in step (2) is inoculated in fermentation tank, 30L ferment tank with 10% inoculum concentration
Culture medium liquid amount is 15L, 37 DEG C of Preliminary fermentation temperature, tank pressure 0.05Mpa, ventilation 1.0vvm, rotating speed 600rpm；Work as fermentation
Adjust rotating speed to 400rpm, ventilation 1.5vvm to 24h, and start to add fed-batch medium, by the feedback regulation of pH value
The flow acceleration of fed-batch medium, control ph first 6 hours in 7.9 to fermentation ends, total fermentation time is 72h；
Described fermentation medium consists of：Glucose 80g/L, yeast extract 20g/L, NH4NO34.5g/L, MgSO4·6H2O0.5g/
L, NaCl 10g/L, CaCl2·6H2O 0.5g/L, FeSO40.01g/L, sodium glutamate 40g/L, NaNO340g/L,MnSO4·
H2O 0.15g/L, pH value 7.5, insufficient section pure water is supplied；
Described fed-batch medium consists of：Glucose 850g/L, sodium chloride 60.0g/L, CaCl2·6H2O 15.0g/L,
MnSO4·H2O10g/L, MgSO4·6H2O 20g/L, pH value 7.9, insufficient section pure water is supplied.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN104017760B (en) * | 2014-06-10 | 2016-07-27 | 河南农业大学 | One strain gamma-polyglutamic acid generating bacterium and the application of tunning thereof |
| CN104694437B (en) * | 2015-03-23 | 2018-04-27 | 领先生物农业股份有限公司 | One bacillus licheniformis and its purposes in gamma-polyglutamic acid is produced |
| CN106282252A (en) * | 2015-06-01 | 2017-01-04 | 山东建筑大学 | A kind of high density fermentation prepares culture medium and the fermentation process thereof of gamma-polyglutamic acid- |
| CN106591190B (en) * | 2016-12-16 | 2019-07-16 | 大连理工大学 | A kind of bacillus and its preparing the application in gamma-polyglutamic acid |
| CN107699594A (en) * | 2017-11-22 | 2018-02-16 | 天津北洋百川生物技术有限公司 | A kind of method that γ polyglutamic acids are produced using bacillus licheniformis |
| CN109161567A (en) * | 2018-10-09 | 2019-01-08 | 四川省食品发酵工业研究设计院 | A method of improving gamma-polyglutamic acid yield |
| CN109182404A (en) * | 2018-10-15 | 2019-01-11 | 天津科技大学 | A method of utilizing stream plus sugar industry gamma-polyglutamic acid when the lichen bacillus ferments |
| CN109988788B (en) * | 2019-04-24 | 2022-11-15 | 武汉骏安生物科技有限公司 | Method for promoting high yield of sodium polyglutamate by bacillus licheniformis |
| CN114456980B (en) * | 2022-02-28 | 2023-08-11 | 天津北洋百川生物技术有限公司 | Gamma-polyglutamic acid high-yield strain and application thereof |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102268465A (en) * | 2011-07-29 | 2011-12-07 | 天津北洋百川生物技术有限公司 | Method for massively producing gamma-poly-glutamic acid |
| CN102367431A (en) * | 2011-11-11 | 2012-03-07 | 中国科学院过程工程研究所 | Bacillus licheniformis and applications thereof |
| CN102719501A (en) * | 2012-06-29 | 2012-10-10 | 天津北洋百川生物技术有限公司 | Method for producing polyglutamic acid |
-
2013
- 2013-12-27 CN CN201310749908.8A patent/CN103695485B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102268465A (en) * | 2011-07-29 | 2011-12-07 | 天津北洋百川生物技术有限公司 | Method for massively producing gamma-poly-glutamic acid |
| CN102367431A (en) * | 2011-11-11 | 2012-03-07 | 中国科学院过程工程研究所 | Bacillus licheniformis and applications thereof |
| CN102719501A (en) * | 2012-06-29 | 2012-10-10 | 天津北洋百川生物技术有限公司 | Method for producing polyglutamic acid |
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