CN114854635A - Fermentation medium of bacillus subtilis and method for producing gamma-polyglutamic acid - Google Patents
Fermentation medium of bacillus subtilis and method for producing gamma-polyglutamic acid Download PDFInfo
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- CN114854635A CN114854635A CN202210539026.8A CN202210539026A CN114854635A CN 114854635 A CN114854635 A CN 114854635A CN 202210539026 A CN202210539026 A CN 202210539026A CN 114854635 A CN114854635 A CN 114854635A
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- 238000000855 fermentation Methods 0.000 title claims abstract description 61
- 230000004151 fermentation Effects 0.000 title claims abstract description 61
- 229920002643 polyglutamic acid Polymers 0.000 title claims abstract description 37
- 244000063299 Bacillus subtilis Species 0.000 title claims abstract description 33
- 235000014469 Bacillus subtilis Nutrition 0.000 title claims abstract description 33
- 238000004519 manufacturing process Methods 0.000 title claims abstract description 15
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 27
- 239000001963 growth medium Substances 0.000 claims abstract description 26
- 239000002609 medium Substances 0.000 claims abstract description 25
- LPUQAYUQRXPFSQ-DFWYDOINSA-M monosodium L-glutamate Chemical compound [Na+].[O-]C(=O)[C@@H](N)CCC(O)=O LPUQAYUQRXPFSQ-DFWYDOINSA-M 0.000 claims abstract description 9
- 239000004223 monosodium glutamate Substances 0.000 claims abstract description 9
- 235000013923 monosodium glutamate Nutrition 0.000 claims abstract description 9
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 16
- 238000009630 liquid culture Methods 0.000 claims description 14
- 239000007788 liquid Substances 0.000 claims description 12
- 238000000034 method Methods 0.000 claims description 10
- 230000004913 activation Effects 0.000 claims description 9
- 238000004321 preservation Methods 0.000 claims description 8
- 239000011780 sodium chloride Substances 0.000 claims description 8
- 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 6
- 239000001888 Peptone Substances 0.000 claims description 6
- 108010080698 Peptones Proteins 0.000 claims description 6
- 235000015278 beef Nutrition 0.000 claims description 6
- 239000008103 glucose Substances 0.000 claims description 6
- 235000019319 peptone Nutrition 0.000 claims description 6
- 239000000843 powder Substances 0.000 claims description 6
- 230000003068 static effect Effects 0.000 claims description 5
- 230000000813 microbial effect Effects 0.000 claims description 4
- 238000009629 microbiological culture Methods 0.000 claims description 4
- 230000010355 oscillation Effects 0.000 claims description 3
- 239000002904 solvent Substances 0.000 claims description 3
- 235000013405 beer Nutrition 0.000 description 12
- 238000002360 preparation method Methods 0.000 description 6
- 230000000052 comparative effect Effects 0.000 description 5
- 230000001954 sterilising effect Effects 0.000 description 5
- LZZYPRNAOMGNLH-UHFFFAOYSA-M Cetrimonium bromide Chemical compound [Br-].CCCCCCCCCCCCCCCC[N+](C)(C)C LZZYPRNAOMGNLH-UHFFFAOYSA-M 0.000 description 4
- 238000012258 culturing Methods 0.000 description 4
- 238000001556 precipitation Methods 0.000 description 4
- GXCLVBGFBYZDAG-UHFFFAOYSA-N N-[2-(1H-indol-3-yl)ethyl]-N-methylprop-2-en-1-amine Chemical compound CN(CCC1=CNC2=C1C=CC=C2)CC=C GXCLVBGFBYZDAG-UHFFFAOYSA-N 0.000 description 3
- 238000007865 diluting Methods 0.000 description 3
- 239000000203 mixture Substances 0.000 description 3
- WHUUTDBJXJRKMK-VKHMYHEASA-N L-glutamic acid Chemical compound OC(=O)[C@@H](N)CCC(O)=O WHUUTDBJXJRKMK-VKHMYHEASA-N 0.000 description 2
- 229920002472 Starch Polymers 0.000 description 2
- 241000209149 Zea Species 0.000 description 2
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 description 2
- 235000002017 Zea mays subsp mays Nutrition 0.000 description 2
- 239000006227 byproduct Substances 0.000 description 2
- 239000003795 chemical substances by application Substances 0.000 description 2
- 235000005822 corn Nutrition 0.000 description 2
- 238000005034 decoration Methods 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 239000008107 starch Substances 0.000 description 2
- 235000019698 starch Nutrition 0.000 description 2
- 241000894006 Bacteria Species 0.000 description 1
- 229920002488 Hemicellulose Polymers 0.000 description 1
- 241000282414 Homo sapiens Species 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 125000000129 anionic group Chemical group 0.000 description 1
- 238000013124 brewing process Methods 0.000 description 1
- 239000001913 cellulose Substances 0.000 description 1
- 229920002678 cellulose Polymers 0.000 description 1
- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 description 1
- 229910000396 dipotassium phosphate Inorganic materials 0.000 description 1
- 235000019797 dipotassium phosphate Nutrition 0.000 description 1
- 235000013399 edible fruits Nutrition 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 230000006353 environmental stress Effects 0.000 description 1
- 239000003337 fertilizer Substances 0.000 description 1
- 238000009472 formulation Methods 0.000 description 1
- 229960002989 glutamic acid Drugs 0.000 description 1
- 150000004676 glycans Chemical class 0.000 description 1
- 229920001519 homopolymer Polymers 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 239000000178 monomer Substances 0.000 description 1
- 231100000252 nontoxic Toxicity 0.000 description 1
- 230000003000 nontoxic effect Effects 0.000 description 1
- 239000002245 particle Substances 0.000 description 1
- 229920001282 polysaccharide Polymers 0.000 description 1
- 239000005017 polysaccharide Substances 0.000 description 1
- LWIHDJKSTIGBAC-UHFFFAOYSA-K potassium phosphate Substances [K+].[K+].[K+].[O-]P([O-])([O-])=O LWIHDJKSTIGBAC-UHFFFAOYSA-K 0.000 description 1
- 239000003755 preservative agent Substances 0.000 description 1
- 230000002335 preservative effect Effects 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 239000003516 soil conditioner Substances 0.000 description 1
- 239000002910 solid waste Substances 0.000 description 1
- 238000003860 storage Methods 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 235000013311 vegetables Nutrition 0.000 description 1
- 239000002699 waste material Substances 0.000 description 1
Classifications
-
- 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
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
-
- 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
- C12P13/00—Preparation of nitrogen-containing organic compounds
- C12P13/02—Amides, e.g. chloramphenicol or polyamides; Imides or polyimides; Urethanes, i.e. compounds comprising N-C=O structural element or polyurethanes
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P60/00—Technologies relating to agriculture, livestock or agroalimentary industries
- Y02P60/80—Food processing, e.g. use of renewable energies or variable speed drives in handling, conveying or stacking
- Y02P60/87—Re-use of by-products of food processing for fodder production
Abstract
The invention relates to a fermentation medium of bacillus subtilis and a method for producing gamma-polyglutamic acid, belonging to the technical field of fermentation. The fermentation medium comprises the following components in parts by weight: 5-10 parts of brewer's grain, 0.15-0.3 part of monosodium glutamate and 10-30 parts of water; the water content of the brewer's grains is 60-65%. The bacillus subtilis can utilize the brewer's grains as a fermentation culture medium to produce the gamma-polyglutamic acid by fermentation, and a new path is searched for the comprehensive utilization of the brewer's grains; and the content of the gamma-polyglutamic acid generated after fermentation is high. The culture medium for producing the gamma-polyglutamic acid by fermentation has simple components, is easy to obtain, has low cost and provides a new way for producing the gamma-polyglutamic acid.
Description
Technical Field
The invention relates to the technical field of fermentation, in particular to a fermentation medium of bacillus subtilis and a method for producing gamma-polyglutamic acid.
Background
Gamma-polyglutamic acid (Gamma-PGA) is a biodegradable nontoxic anionic homopolymer consisting of D-and L-glutamic acid monomers, and can be widely applied to the agricultural field as a water-retaining agent, a fruit, vegetable and flower storage preservative, a growth and development promoter, a fertilizer synergist, an environmental stress resistance agent, a soil conditioner and the like.
Beer lees, the most important by-product produced in the beer brewing process, accounts for 85% of all by-products in beer production. The beer lees is the undissolved residue of malt and auxiliary materials in the saccharification process, mainly comprises seed coats and shells of the malt, a small amount of incompletely dissolved starch endosperm particles and the like, and mainly comprises non-starch polysaccharides (cellulose and hemicellulose; 30-50%) and proteins (19-30%) from the chemical composition. It is estimated that the annual average production of brewery mash worldwide is 3900 ten thousand tons. Generally, 20kg of wet brewery mash is produced for every 100L of beer produced; in China, each ton of malt can produce 1.1-1.3 tons of wet brewer's grain with 75% -80% of water content. At present, the beer lees are mainly utilized as animal feed, the annual average yield of the produced beer lees is increased along with the annual increase of the global beer output, the feed market demand is limited, and therefore, the redundant beer lees are generally directly discharged as solid waste, which wastes resources and pollutes the environment. With the enhancement of the environmental awareness of human beings, the development and utilization of the brewer's grains become a research hotspot. No method for producing gamma-polyglutamic acid by using brewer's grains nor a precedent for which strain can be used for fermenting gamma-polyglutamic acid by using brewer's grains has been found in the prior art.
Disclosure of Invention
The invention aims to provide a fermentation medium of bacillus subtilis and a method for producing gamma-polyglutamic acid.
In order to achieve the above object, the present invention provides the following technical solutions:
the invention also provides a fermentation medium of the bacillus subtilis, which comprises the following components in parts by weight:
5-10 parts of brewer's grain, 0.15-0.3 part of monosodium glutamate and 10-30 parts of water;
the water content of the brewer's grains is 60-65%.
The invention also provides a method for producing gamma-polyglutamic acid by using the fermentation culture medium, which comprises the following steps:
(1) inoculating bacillus subtilis into a liquid culture medium for activated culture to obtain a seed solution;
(2) inoculating the seed solution into the fermentation culture medium for fermentation culture to obtain gamma-polyglutamic acid;
the bacillus subtilis is bacillus subtilis YB 18;
the bacillus subtilis YB18 is preserved in the China general microbiological culture Collection center (CGMCC), the address is No. 3 of West Lu No.1 of the morning-West Chen in the Korean-Yang district of Beijing, the microbial research institute of the Chinese academy of sciences, the preservation number is CGMCC No.17642, the bacillus subtilis is named by classification, and the preservation time is 2019, 4 months and 28 days;
the initial viable count contained in the inoculated seed liquid is 1.0-1.5 multiplied by 10 8 cfu/mL; the volume ratio of the seed liquid to the fermentation medium is 4-5: 2500.
Preferably, the liquid culture medium in the step (1) takes water as a solvent and comprises the following components in concentration:
8-12 g/L of peptone, 2.5-3.5 g/L of beef extract powder, 18-22 g/L of glucose and 4-6 g/L of sodium chloride.
Preferably, the temperature of the activation culture in the step (1) is 35-39 ℃;
the activation culture time is 16-20 h;
the activation culture is shaking culture;
the rotating speed of the oscillation is 140-160 rpm.
Preferably, the temperature of the fermentation culture in the step (2) is 35-39 ℃;
the fermentation culture time is 45-72 h;
the fermentation culture is static culture.
The invention provides bacillus subtilis, a fermentation culture medium thereof and a method for producing gamma-polyglutamic acid. The strain and the production method of the invention have the following advantages:
(1) the bacillus subtilis can utilize the brewer's grains as a fermentation culture medium to produce the gamma-polyglutamic acid by fermentation, and a new path is searched for the comprehensive utilization of the brewer's grains; and the content of the gamma-polyglutamic acid generated after fermentation is high.
(2) The culture medium for producing the gamma-polyglutamic acid by fermentation has simple components, is easy to obtain, has low cost and provides a new way for producing the gamma-polyglutamic acid.
Deposit description
The Bacillus subtilis YB18 is preserved in the China general microbiological culture Collection center (CGMCC), the address is No. 3 of West Lu No.1 of the morning-West Chen in the south-Yang district of Beijing, the microbial research institute of the Chinese academy of sciences, the preservation number is CGMCC No.17642, the Bacillus subtilis is named by classification, and the preservation time is 2019, 4 months and 28 days.
Detailed Description
The invention provides a fermentation medium of bacillus subtilis, which comprises the following components in parts by weight:
5-10 parts of brewer's grain, preferably 7.5 parts;
0.15-0.3 part of monosodium glutamate, preferably 0.225 part;
10-30 parts of water, preferably 20 parts;
the water content of the brewer's grains is 60-65%, and the preferable water content is 62.5%.
The invention also provides a method for producing gamma-polyglutamic acid by using the fermentation culture medium, which comprises the following steps:
(1) inoculating bacillus subtilis into a liquid culture medium for activated culture to obtain a seed solution;
(2) inoculating the seed solution into the fermentation culture medium for fermentation culture to obtain gamma-polyglutamic acid;
the bacillus subtilis is bacillus subtilis YB 18;
the Bacillus subtilis YB18 is preserved in the China general microbiological culture Collection center (CGMCC), the address is No. 3 of West Lu No.1 of the morning-West Chen in the morning-Yang district of Beijing, the microbial research institute of the Chinese academy of sciences, the preservation number is CGMCC No.17642, the Bacillus subtilis is named by classification, and the preservation time is 2019, 4 months and 28 days;
the initial viable count contained in the inoculated seed liquid is 1.0-1.5 multiplied by 10 8 cfu/mL, preferably 1.25X 10 8 cfu/mL; the volume ratio of the seed liquid to the fermentation medium is 4-5: 2500, and preferably 4.5: 2500.
In the invention, the liquid culture medium in the step (1) takes water as a solvent and comprises the following components in concentration:
peptone is 8-12 g/L, preferably 10 g/L;
2.5-3.5 g/L of beef extract powder, preferably 3 g/L;
18-22 g/L of glucose, preferably 20 g/L;
4-6 g/L of sodium chloride, preferably 5 g/L.
In the invention, the temperature of the activation culture in the step (1) is 35-39 ℃, preferably 36-38 ℃, and more preferably 37 ℃; the activation culture time is 16-20 h, preferably 17-19 h, and further preferably 18 h; the activation culture is shaking culture; the rotating speed of the oscillation is 140-160 rpm, and preferably 150 rpm.
In the invention, the temperature of the fermentation culture in the step (2) is 35-39 ℃, preferably 36-38 ℃, and more preferably 37 ℃; the fermentation culture time is 45-72 h, preferably 50-67 h, further preferably 55-62 h, and further preferably 58.5 h; the fermentation culture is static culture.
The embodiments of the present invention will be described in detail with reference to the following examples, but they should not be construed as limiting the scope of the present invention.
Example 1
The preparation method of each liter of liquid culture medium comprises the following steps: taking 800mL of water, then adding 8g of peptone, 3.5g of beef extract powder, 20g of glucose and 6g of sodium chloride, diluting to 1L with water, and sterilizing at 121 ℃ for 20min to obtain a liquid culture medium.
The preparation method of each liter of fermentation medium comprises the following steps: taking 3000mL of water, adding 800g of beer lees with the water content of 60% and 30g of monosodium glutamate, and sterilizing at 121 ℃ for 20min to obtain the fermentation medium.
Selecting slant strain of Bacillus subtilis YB18, inoculating in liquid culture medium containing 50mL, shake culturing at 35 deg.C and 160rpm for 18h to obtain viable count of 1.20 × 10 8 cfu/mL of seed solution. Inoculating 4mL of seed liquid into 2500mL of fermentation medium, performing static culture at 35 ℃ for 60h, and detecting the yield of the gamma-polyglutamic acid by using a CTAB precipitation method, wherein the yield is 1000g of brewer's grain, so that 30.75g of gamma-polyglutamic acid can be obtained by fermentation.
Example 2
The preparation method of each liter of liquid culture medium comprises the following steps: taking 800mL of water, then adding 12g of peptone, 3g of beef extract powder, 18g of glucose and 4g of sodium chloride, diluting to 1L with water, and sterilizing at 121 ℃ for 20min to obtain a liquid culture medium.
The preparation method of each liter of fermentation medium comprises the following steps: and (3) taking 5000mL of water, adding 1250g of brewer's grains with the water content of 65% and 50g of monosodium glutamate, and sterilizing at 121 ℃ for 20min to obtain the fermentation medium.
Selecting slant strain of Bacillus subtilis YB18, inoculating in liquid culture medium containing 50mL, shake culturing at 39 deg.C and 140rpm for 16h to obtain viable count of 1.0 × 10 8 cfu/mL of seed solution. Inoculating 5mL of seed liquid into 2500mL of fermentation medium, statically culturing at 39 ℃ for 45h, and detecting the yield of the gamma-polyglutamic acid by a CTAB precipitation method, wherein the yield is 1000g of brewer's grain, so that 31.25g of gamma-polyglutamic acid can be obtained by fermentation.
Example 3
The preparation method of each liter of liquid culture medium comprises the following steps: taking 800mL of water, then adding 10g of peptone, 2.5g of beef extract powder, 22g of glucose and 5g of sodium chloride, diluting to 1L with water, and sterilizing at 121 ℃ for 20min to obtain a liquid culture medium.
The preparation method of each liter of fermentation medium comprises the following steps: 5000mL of water is taken, 5000g of beer lees with the water content of 62% and 75g of monosodium glutamate are added, and the mixture is sterilized for 20min at 121 ℃ to obtain the fermentation medium.
Selecting slant strain of Bacillus subtilis YB18, inoculating in liquid culture medium containing 50mL, shake culturing at 37 deg.C and 150rpm for 20 hr to obtain viable bacteria number of 1.5 × 10 8 cfu/mL of seed solution. Inoculating 4mL of seed liquid into 2500mL of fermentation medium, performing static culture at 38 ℃ for 72h, and detecting the yield of the gamma-polyglutamic acid to be 1000g of brewer's grain by using a CTAB precipitation method to ferment so as to obtain 33.45g of gamma-polyglutamic acid.
Comparative example 1
The method of comparative example 1 of the present application was set up according to the method of example 1, and unlike example 1, the fermentation medium formulation of comparative example 1 was: 0.5L/L of yellow water, 50g/L of reducing sugar content of corn saccharification liquid, 60g/L of monosodium glutamate and 9g/L, K g of sodium chloride 2 HPO 4 4g/L and pH value of 5. After the culture of the fermentation medium, the yield of the gamma-polyglutamic acid detected by a CTAB precipitation method is 14.51 g/L.
Application example 1
The fermentation media of examples 1 to 3 and comparative example 1 were subjected to cost calculation, and capital spent per 100g of gamma-polyglutamic acid produced by fermentation was compared. The results are shown in Table 1.
TABLE 1100g raw Material cost for Gamma-polyglutamic acid production
| Example 1 | Example 2 | Example 3 | Comparative example 1 | |
| Cost/dollar | 3.356084 | 3.3024 | 3.085201 | 9.264483 |
The beer grains are calculated according to 0.0006 yuan/g; the monosodium glutamate is calculated according to 0.01152 yuan/g; calculating the yellow water according to 1.001 yuan/mL; calculating the corn saccharification liquid according to 0.0018 yuan/mL; calculating the sodium chloride according to 0.0158 yuan/g; dipotassium phosphate was calculated at 0.035 yuan/g.
As can be seen from the above examples, the present invention provides a fermentation medium of Bacillus subtilis and a method for producing gamma-polyglutamic acid. The bacillus subtilis can utilize the brewer's grains as a fermentation culture medium to produce the gamma-polyglutamic acid by fermentation, and a new path is searched for the comprehensive utilization of the brewer's grains; and the content of the gamma-polyglutamic acid generated after fermentation is high. The culture medium for producing the gamma-polyglutamic acid by fermentation has simple components, is easy to obtain, has low cost and provides a new way for producing the gamma-polyglutamic acid.
The foregoing is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, various modifications and decorations can be made without departing from the principle of the present invention, and these modifications and decorations should also be regarded as the protection scope of the present invention.
Claims (5)
1. The fermentation medium of the bacillus subtilis is characterized by comprising the following components in parts by mass:
5-10 parts of brewer's grain, 0.15-0.3 part of monosodium glutamate and 10-30 parts of water;
the water content of the brewer's grains is 60-65%.
2. The method for producing gamma-polyglutamic acid using the fermentation medium of claim 2, comprising the steps of:
(1) inoculating bacillus subtilis into a liquid culture medium for activated culture to obtain a seed solution;
(2) inoculating the seed solution into the fermentation culture medium of claim 1 for fermentation culture to obtain gamma-polyglutamic acid;
the bacillus subtilis is bacillus subtilis YB 18;
the Bacillus subtilis YB18 is preserved in the China general microbiological culture Collection center (CGMCC), the address is No. 3 of West Lu No.1 of the morning-West Chen in the morning-Yang district of Beijing, the microbial research institute of the Chinese academy of sciences, the preservation number is CGMCC No.17642, the Bacillus subtilis is named by classification, and the preservation time is 2019, 4 months and 28 days;
the initial viable count contained in the inoculated seed liquid is 1.0-1.5 multiplied by 10 8 cfu/mL; the volume ratio of the seed liquid to the fermentation medium is 4-5: 2500.
3. The method according to claim 2, wherein the liquid medium of step (1) comprises the following components in the following concentrations in water as a solvent:
8-12 g/L of peptone, 2.5-3.5 g/L of beef extract powder, 18-22 g/L of glucose and 4-6 g/L of sodium chloride.
4. The method according to claim 3, wherein the temperature of the activation culture in the step (1) is 35-39 ℃;
the activation culture time is 16-20 h;
the activation culture is shaking culture;
the rotating speed of the oscillation is 140-160 rpm.
5. The method according to claim 4, wherein the temperature of the fermentation culture in the step (2) is 35-39 ℃;
the fermentation culture time is 45-72 h;
the fermentation culture is static culture.
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