CN117965654B - Method for shortening fermentation period of L-valine - Google Patents
Method for shortening fermentation period of L-valine Download PDFInfo
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- CN117965654B CN117965654B CN202410389221.6A CN202410389221A CN117965654B CN 117965654 B CN117965654 B CN 117965654B CN 202410389221 A CN202410389221 A CN 202410389221A CN 117965654 B CN117965654 B CN 117965654B
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- valine
- chloride
- shortening
- escherichia coli
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- 238000000855 fermentation Methods 0.000 title claims abstract description 210
- 230000004151 fermentation Effects 0.000 title claims abstract description 210
- KZSNJWFQEVHDMF-BYPYZUCNSA-N L-valine Chemical compound CC(C)[C@H](N)C(O)=O KZSNJWFQEVHDMF-BYPYZUCNSA-N 0.000 title claims abstract description 191
- 229960004295 valine Drugs 0.000 title claims abstract description 95
- 238000000034 method Methods 0.000 title claims abstract description 42
- 238000004904 shortening Methods 0.000 title claims abstract description 19
- 239000000203 mixture Substances 0.000 claims abstract description 76
- 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 abstract description 30
- 239000008103 glucose Substances 0.000 claims abstract description 30
- 229910052751 metal Inorganic materials 0.000 claims abstract description 28
- 239000002184 metal Substances 0.000 claims abstract description 28
- FVAUCKIRQBBSSJ-UHFFFAOYSA-M sodium iodide Chemical compound [Na+].[I-] FVAUCKIRQBBSSJ-UHFFFAOYSA-M 0.000 claims abstract description 21
- YBJHBAHKTGYVGT-ZKWXMUAHSA-N (+)-Biotin Chemical compound N1C(=O)N[C@@H]2[C@H](CCCCC(=O)O)SC[C@@H]21 YBJHBAHKTGYVGT-ZKWXMUAHSA-N 0.000 claims abstract description 14
- DFPAKSUCGFBDDF-UHFFFAOYSA-N Nicotinamide Chemical compound NC(=O)C1=CC=CN=C1 DFPAKSUCGFBDDF-UHFFFAOYSA-N 0.000 claims abstract description 14
- 241001052560 Thallis Species 0.000 claims abstract description 14
- JIAARYAFYJHUJI-UHFFFAOYSA-L zinc dichloride Chemical compound [Cl-].[Cl-].[Zn+2] JIAARYAFYJHUJI-UHFFFAOYSA-L 0.000 claims abstract description 14
- 238000009423 ventilation Methods 0.000 claims abstract description 11
- 239000001763 2-hydroxyethyl(trimethyl)azanium Substances 0.000 claims abstract description 7
- 235000019743 Choline chloride Nutrition 0.000 claims abstract description 7
- 235000000638 D-biotin Nutrition 0.000 claims abstract description 7
- 239000011665 D-biotin Substances 0.000 claims abstract description 7
- 229910021586 Nickel(II) chloride Inorganic materials 0.000 claims abstract description 7
- 239000004115 Sodium Silicate Substances 0.000 claims abstract description 7
- KGBXLFKZBHKPEV-UHFFFAOYSA-N boric acid Chemical compound OB(O)O KGBXLFKZBHKPEV-UHFFFAOYSA-N 0.000 claims abstract description 7
- 239000004327 boric acid Substances 0.000 claims abstract description 7
- AIYUHDOJVYHVIT-UHFFFAOYSA-M caesium chloride Chemical compound [Cl-].[Cs+] AIYUHDOJVYHVIT-UHFFFAOYSA-M 0.000 claims abstract description 7
- SGMZJAMFUVOLNK-UHFFFAOYSA-M choline chloride Chemical compound [Cl-].C[N+](C)(C)CCO SGMZJAMFUVOLNK-UHFFFAOYSA-M 0.000 claims abstract description 7
- 229960003178 choline chloride Drugs 0.000 claims abstract description 7
- GVPFVAHMJGGAJG-UHFFFAOYSA-L cobalt dichloride Chemical compound [Cl-].[Cl-].[Co+2] GVPFVAHMJGGAJG-UHFFFAOYSA-L 0.000 claims abstract description 7
- ORTQZVOHEJQUHG-UHFFFAOYSA-L copper(II) chloride Chemical compound Cl[Cu]Cl ORTQZVOHEJQUHG-UHFFFAOYSA-L 0.000 claims abstract description 7
- BVTBRVFYZUCAKH-UHFFFAOYSA-L disodium selenite Chemical compound [Na+].[Na+].[O-][Se]([O-])=O BVTBRVFYZUCAKH-UHFFFAOYSA-L 0.000 claims abstract description 7
- QMMRZOWCJAIUJA-UHFFFAOYSA-L nickel dichloride Chemical compound Cl[Ni]Cl QMMRZOWCJAIUJA-UHFFFAOYSA-L 0.000 claims abstract description 7
- 229960003966 nicotinamide Drugs 0.000 claims abstract description 7
- 235000005152 nicotinamide Nutrition 0.000 claims abstract description 7
- 239000011570 nicotinamide Substances 0.000 claims abstract description 7
- 230000008569 process Effects 0.000 claims abstract description 7
- DVMZCYSFPFUKKE-UHFFFAOYSA-K scandium chloride Chemical compound Cl[Sc](Cl)Cl DVMZCYSFPFUKKE-UHFFFAOYSA-K 0.000 claims abstract description 7
- 235000009518 sodium iodide Nutrition 0.000 claims abstract description 7
- 235000015393 sodium molybdate Nutrition 0.000 claims abstract description 7
- 239000011684 sodium molybdate Substances 0.000 claims abstract description 7
- TVXXNOYZHKPKGW-UHFFFAOYSA-N sodium molybdate (anhydrous) Chemical compound [Na+].[Na+].[O-][Mo]([O-])(=O)=O TVXXNOYZHKPKGW-UHFFFAOYSA-N 0.000 claims abstract description 7
- 235000015921 sodium selenite Nutrition 0.000 claims abstract description 7
- 239000011781 sodium selenite Substances 0.000 claims abstract description 7
- 229960001471 sodium selenite Drugs 0.000 claims abstract description 7
- NTHWMYGWWRZVTN-UHFFFAOYSA-N sodium silicate Chemical compound [Na+].[Na+].[O-][Si]([O-])=O NTHWMYGWWRZVTN-UHFFFAOYSA-N 0.000 claims abstract description 7
- 229910052911 sodium silicate Inorganic materials 0.000 claims abstract description 7
- 235000005074 zinc chloride Nutrition 0.000 claims abstract description 7
- 239000011592 zinc chloride Substances 0.000 claims abstract description 7
- 241000588724 Escherichia coli Species 0.000 claims description 25
- 239000007788 liquid Substances 0.000 claims description 22
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 22
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 claims description 20
- 239000002904 solvent Substances 0.000 claims description 20
- 239000001963 growth medium Substances 0.000 claims description 18
- 239000002609 medium Substances 0.000 claims description 16
- 238000011218 seed culture Methods 0.000 claims description 12
- 239000000843 powder Substances 0.000 claims description 11
- 240000004808 Saccharomyces cerevisiae Species 0.000 claims description 10
- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 claims description 10
- 229910052943 magnesium sulfate Inorganic materials 0.000 claims description 10
- 235000019341 magnesium sulphate Nutrition 0.000 claims description 10
- YWYZEGXAUVWDED-UHFFFAOYSA-N triammonium citrate Chemical compound [NH4+].[NH4+].[NH4+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O YWYZEGXAUVWDED-UHFFFAOYSA-N 0.000 claims description 10
- 229910000402 monopotassium phosphate Inorganic materials 0.000 claims description 9
- 235000019796 monopotassium phosphate Nutrition 0.000 claims description 9
- PJNZPQUBCPKICU-UHFFFAOYSA-N phosphoric acid;potassium Chemical compound [K].OP(O)(O)=O PJNZPQUBCPKICU-UHFFFAOYSA-N 0.000 claims description 6
- KWIUHFFTVRNATP-UHFFFAOYSA-N Betaine Natural products C[N+](C)(C)CC([O-])=O KWIUHFFTVRNATP-UHFFFAOYSA-N 0.000 claims description 5
- KWIUHFFTVRNATP-UHFFFAOYSA-O N,N,N-trimethylglycinium Chemical compound C[N+](C)(C)CC(O)=O KWIUHFFTVRNATP-UHFFFAOYSA-O 0.000 claims description 5
- 229960003237 betaine Drugs 0.000 claims description 5
- 238000012136 culture method Methods 0.000 claims description 2
- 238000002360 preparation method Methods 0.000 claims description 2
- 238000011081 inoculation Methods 0.000 claims 1
- 238000006243 chemical reaction Methods 0.000 abstract description 24
- 238000004519 manufacturing process Methods 0.000 abstract description 21
- 239000011573 trace mineral Substances 0.000 abstract description 21
- 235000013619 trace mineral Nutrition 0.000 abstract description 21
- 230000000694 effects Effects 0.000 abstract description 9
- 102000004190 Enzymes Human genes 0.000 abstract description 7
- 108090000790 Enzymes Proteins 0.000 abstract description 7
- 238000009825 accumulation Methods 0.000 abstract description 5
- 230000009286 beneficial effect Effects 0.000 abstract description 5
- 230000000813 microbial effect Effects 0.000 abstract description 5
- 230000007269 microbial metabolism Effects 0.000 abstract description 4
- 230000037358 bacterial metabolism Effects 0.000 abstract description 3
- 235000010338 boric acid Nutrition 0.000 abstract 1
- 235000019794 sodium silicate Nutrition 0.000 abstract 1
- 230000000052 comparative effect Effects 0.000 description 31
- 238000012795 verification Methods 0.000 description 31
- 239000002253 acid Substances 0.000 description 15
- 230000012010 growth Effects 0.000 description 13
- 239000000243 solution Substances 0.000 description 10
- 229910052761 rare earth metal Inorganic materials 0.000 description 7
- 150000002910 rare earth metals Chemical class 0.000 description 7
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 6
- 238000002835 absorbance Methods 0.000 description 5
- 239000003795 chemical substances by application Substances 0.000 description 5
- 238000003786 synthesis reaction Methods 0.000 description 5
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 4
- LRHPLDYGYMQRHN-UHFFFAOYSA-N N-Butanol Chemical group CCCCO LRHPLDYGYMQRHN-UHFFFAOYSA-N 0.000 description 4
- 230000008901 benefit Effects 0.000 description 4
- GNSKLFRGEWLPPA-UHFFFAOYSA-M potassium dihydrogen phosphate Chemical compound [K+].OP(O)([O-])=O GNSKLFRGEWLPPA-UHFFFAOYSA-M 0.000 description 4
- 239000000523 sample Substances 0.000 description 4
- 230000001580 bacterial effect Effects 0.000 description 3
- 230000015572 biosynthetic process Effects 0.000 description 3
- 239000002054 inoculum Substances 0.000 description 3
- LWIHDJKSTIGBAC-UHFFFAOYSA-K potassium phosphate Substances [K+].[K+].[K+].[O-]P([O-])([O-])=O LWIHDJKSTIGBAC-UHFFFAOYSA-K 0.000 description 3
- 238000012360 testing method Methods 0.000 description 3
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 description 2
- LCTONWCANYUPML-UHFFFAOYSA-N Pyruvic acid Chemical compound CC(=O)C(O)=O LCTONWCANYUPML-UHFFFAOYSA-N 0.000 description 2
- 229960000583 acetic acid Drugs 0.000 description 2
- 229940024606 amino acid Drugs 0.000 description 2
- 150000001413 amino acids Chemical class 0.000 description 2
- 230000010261 cell growth Effects 0.000 description 2
- 229910000365 copper sulfate Inorganic materials 0.000 description 2
- ARUVKPQLZAKDPS-UHFFFAOYSA-L copper(II) sulfate Chemical compound [Cu+2].[O-][S+2]([O-])([O-])[O-] ARUVKPQLZAKDPS-UHFFFAOYSA-L 0.000 description 2
- 238000010790 dilution Methods 0.000 description 2
- 239000012895 dilution Substances 0.000 description 2
- 239000012153 distilled water Substances 0.000 description 2
- 239000003480 eluent Substances 0.000 description 2
- 239000012362 glacial acetic acid Substances 0.000 description 2
- 230000006872 improvement Effects 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 238000005259 measurement Methods 0.000 description 2
- 230000002503 metabolic effect Effects 0.000 description 2
- 230000004060 metabolic process Effects 0.000 description 2
- 230000007065 protein hydrolysis Effects 0.000 description 2
- 239000002994 raw material Substances 0.000 description 2
- 238000005070 sampling Methods 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 241000894006 Bacteria Species 0.000 description 1
- QIVBCDIJIAJPQS-VIFPVBQESA-N L-tryptophane Chemical compound C1=CC=C2C(C[C@H](N)C(O)=O)=CNC2=C1 QIVBCDIJIAJPQS-VIFPVBQESA-N 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- QIVBCDIJIAJPQS-UHFFFAOYSA-N Tryptophan Natural products C1=CC=C2C(CC(N)C(O)=O)=CNC2=C1 QIVBCDIJIAJPQS-UHFFFAOYSA-N 0.000 description 1
- KZSNJWFQEVHDMF-UHFFFAOYSA-N Valine Chemical compound CC(C)C(N)C(O)=O KZSNJWFQEVHDMF-UHFFFAOYSA-N 0.000 description 1
- 241000212749 Zesius chrysomallus Species 0.000 description 1
- 150000007513 acids Chemical class 0.000 description 1
- 238000005273 aeration Methods 0.000 description 1
- 230000003698 anagen phase Effects 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 1
- 150000005693 branched-chain amino acids Chemical class 0.000 description 1
- 238000004587 chromatography analysis Methods 0.000 description 1
- 239000013078 crystal Substances 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000018109 developmental process Effects 0.000 description 1
- 239000012470 diluted sample Substances 0.000 description 1
- 238000007865 diluting Methods 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 238000001035 drying Methods 0.000 description 1
- 238000005265 energy consumption Methods 0.000 description 1
- 239000003797 essential amino acid Substances 0.000 description 1
- 235000020776 essential amino acid Nutrition 0.000 description 1
- 238000001704 evaporation Methods 0.000 description 1
- 230000008020 evaporation Effects 0.000 description 1
- 230000002349 favourable effect Effects 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 238000010438 heat treatment Methods 0.000 description 1
- 125000005842 heteroatom Chemical group 0.000 description 1
- 239000002207 metabolite Substances 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 230000007935 neutral effect Effects 0.000 description 1
- FEMOMIGRRWSMCU-UHFFFAOYSA-N ninhydrin Chemical group C1=CC=C2C(=O)C(O)(O)C(=O)C2=C1 FEMOMIGRRWSMCU-UHFFFAOYSA-N 0.000 description 1
- 230000009965 odorless effect Effects 0.000 description 1
- 150000007524 organic acids Chemical class 0.000 description 1
- 239000001301 oxygen Substances 0.000 description 1
- 229910052760 oxygen Inorganic materials 0.000 description 1
- 238000004816 paper chromatography Methods 0.000 description 1
- 230000002035 prolonged effect Effects 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 229940107700 pyruvic acid Drugs 0.000 description 1
- 238000005507 spraying Methods 0.000 description 1
- 238000003892 spreading Methods 0.000 description 1
- 230000007480 spreading Effects 0.000 description 1
- 230000002194 synthesizing effect Effects 0.000 description 1
Classifications
-
- 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
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Landscapes
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
The invention is suitable for the technical field of microbial fermentation, and provides a method for shortening the fermentation period of L-valine, which comprises the following steps of adding a trace element composition into a fermentation medium: sodium selenite, boric acid, sodium silicate, sodium iodide, D-biotin, nicotinamide and choline chloride; rare metal compositions were also added: nickel chloride, cobalt chloride, copper chloride, zinc chloride, sodium molybdate, scandium chloride and cesium chloride; and controlling ventilation and glucose concentration in the fermentation culture process. In conclusion, the trace element composition is added into the fermentation medium, so that the thalli grow and propagate rapidly, and the fermentation period is shortened; by adding the rare metal composition into the fermentation medium, the bacterial metabolism enzyme activity is improved, so that the microbial metabolism is more beneficial to the production and accumulation of L-valine, the fermentation conversion rate of the L-valine is improved, and the production cost is reduced.
Description
Technical Field
The invention relates to the technical field of microbial fermentation, in particular to a method for shortening the fermentation period of L-valine.
Background
L-valine is chemically named as alpha-amino-3-methylbutanoic acid, odorless, bitter in taste, white monoclinic crystal or crystalline powder. L-valine is neutral amino acid, the aqueous solution of which is weak acid, belongs to branched chain amino acid, is one of essential amino acids of human body, and has wide application in the industries of medicines, foods and animal feeds.
The L-valine can be produced by chemical synthesis, protein hydrolysis and microbial fermentation. The chemical synthesis method and the protein hydrolysis method are easy to cause serious pollution to the environment when producing the L-valine, have huge energy consumption, high production cost and low efficiency, and are not suitable for the output production of the L-valine. The microbial fermentation method is a main production mode of L-valine at present, and is a method for directly fermenting and synthesizing the L-valine by taking glucose as a raw material by utilizing the biological metabolism of the microorganism; the method has the advantages of low cost of raw materials, mild and easily controlled reaction conditions, small pollution and the like, and becomes a main production method of L-valine at home and abroad.
However, in the process of producing L-valine by a microbial fermentation method, the cost is mainly influenced by the fermentation level, and the method is mainly characterized by low sugar-acid conversion rate, has higher production cost and is unfavorable for the green development of industry.
Patent publication No. CN114875089A discloses a method for improving the fermentation efficiency of L-valine, which promotes the accumulation of a large amount of pyruvic acid by adding a small amount of tryptophan into a fermentation medium, thereby improving the acid yield of L-valine and finally enabling the conversion rate to be about 46%, and the conversion rate is still to be improved although the conversion rate of L-valine is improved to a certain extent.
Disclosure of Invention
In view of the above, the invention provides a method for shortening the fermentation period of L-valine, which is characterized in that trace element composition and rare metal composition are added to ferment and prepare L-valine, so that thalli grow and reproduce rapidly, the fermentation period is shortened, microbial metabolism is more beneficial to producing and accumulating L-valine, and the fermentation conversion rate is improved, thereby reducing the production cost.
The technical scheme of the invention is realized as follows:
the invention provides a method for shortening the fermentation period of L-valine, which takes escherichia coli as a strain, and adds a microelement composition and a rare metal composition into a fermentation culture medium for fermentation culture;
Wherein, the microelement composition takes water as a solvent and also comprises the following components with the concentration: 1-3g/L of sodium selenite, 0.2-0.4g/L of boric acid, 10-12g/L of sodium silicate, 5-8g/L of sodium iodide, 2-5g/L of D-biotin, 20-25g/L of nicotinamide and 4-6g/L of choline chloride.
Wherein, rare metal composition uses water as solvent, still include the component of following concentration: 0.3-0.5g/L of nickel chloride, 3-6g/L of cobalt chloride, 10-15g/L of copper chloride, 5-8g/L of zinc chloride, 5-7g/L of sodium molybdate, 0.2-0.5g/L of scandium chloride and 0.2-0.4g/L of cesium chloride.
The microelement composition is added to enable the escherichia coli to grow rapidly, and the growth lag phase of the thalli is shortened, so that the fermentation period is shortened, and the fermentation production efficiency is improved.
The rare metal composition is added, so that the microbial metabolism enzyme activity is improved, the production and accumulation of L-valine are promoted, the sugar acid conversion rate is effectively improved, and the production cost is reduced.
Preferably, the fermentation medium takes water as a solvent, the microelement composition is 1-2g/L, and the rare metal composition is 0.2-0.5g/L; the composition also comprises the following basic components in concentration: 5-8g/L of yeast powder, 0.6-0.8g/L of betaine, 10-15g/L of glucose, 0.2-0.5g/L of magnesium sulfate, 1-2g/L of dipotassium hydrogen phosphate, 1-2g/L of potassium dihydrogen phosphate and 2-4g/L of ammonium citrate.
Preferably, the fermentation culture method comprises the following steps: inoculating the escherichia coli seed liquid into a fermentation culture medium, controlling the temperature to be 35-38 ℃, the pH value to be 6.9-7.1, the pressure to be 0.03-0.05MPa, the ventilation rate to be 0.5-0.8 m air/m fermentation liquid/min, simultaneously feeding 60-65% (w/v) glucose solution, controlling the glucose concentration in the fermentation liquid to be 0.03-0.05% (w/v), and performing fermentation culture, wherein the fermentation is finished when the L-valine content in the fermentation liquid is 7.5-8.5%; if the L-valine content in the fermentation liquor is more than 8.5%, the fermentation liquor is easy to crystallize, so that the pipeline is blocked to influence the fermentation, and the fermentation is finished within the range, thereby being more beneficial to fermentation production. In the present invention, the fermentation period of L-valine is 23-24 hours from the beginning to the end of fermentation.
Preferably, the E.coli seed solution is inoculated in an amount of 13-16% by volume of the fermentation medium.
Preferably, the preparation method of the escherichia coli seed liquid comprises the following steps: and inoculating the escherichia coli into a seed culture medium for seed culture until the OD 610nm value of the thalli is 10-12, so as to obtain the escherichia coli seed liquid. The escherichia coli seed liquid with the thallus OD 610nm value is in a growth log phase, and has the advantage of rapid growth and propagation.
Preferably, the seed culture medium takes water as a solvent, and further comprises the following components in concentration: 10-12g/L of yeast powder, 25-30g/L of glucose, 0.5-0.8g/L of magnesium sulfate, 4-6g/L of dipotassium hydrogen phosphate, 4-6g/L of potassium dihydrogen phosphate and 4-7g/L of ammonium citrate.
Preferably, the culture temperature of the escherichia coli seed solution is 35-37 ℃, the pH value is 6.9-7.0, and the pressure is 0.03-0.05MPa.
The type of the strain of the E.coli is not particularly limited, and E.coli conventionally used in the art for fermentatively producing L-valine may be used. The sources of the components in the medium and composition of the present invention are not particularly limited and may be purchased through conventional commercial sources. In the present invention, the unit of mass volume concentration (w/v) defaults to g/dL unless otherwise specified.
Compared with the prior art, the method for shortening the fermentation period of the L-valine has the following beneficial effects:
1. the trace element composition is added into the fermentation medium to enable the thalli to grow and reproduce rapidly, so that the fermentation period is shortened;
2. The rare metal composition is added into the fermentation medium, so that the bacterial metabolism enzyme activity is improved, and the microbial metabolism is more beneficial to the production and accumulation of L-valine, thereby improving the fermentation conversion rate of L-valine and reducing the production cost;
3. the conditions of the fermentation culture process are adapted to the improvement of the fermentation medium of the invention by controlling the ventilation in the fermentation process and the glucose concentration in the fermentation broth.
Drawings
In order to more clearly illustrate the embodiments of the invention or the technical solutions in the prior art, the drawings that are required in the embodiments or the description of the prior art will be briefly described, it being obvious that the drawings in the following description are only some embodiments of the invention, and that other drawings may be obtained according to these drawings without inventive effort for a person skilled in the art.
FIG. 1 shows the growth of the cells OD 610nm of example 2 during different fermentation periods;
FIG. 2 is a graph showing the results of measurement of L-valine content in example 2 at various fermentation periods;
FIG. 3 shows the growth of the cells OD 610nm of the comparative example 1 at different fermentation periods;
FIG. 4 is a graph showing the results of measurement of L-valine content in comparative example 1 at various fermentation periods.
Detailed Description
The following description of the embodiments of the present invention will clearly and fully describe the technical aspects of the embodiments of the present invention, and it is apparent that the described embodiments are only some embodiments of the present invention, not all embodiments. All other embodiments, which can be made by those skilled in the art based on the embodiments of the present invention without making any inventive effort, are intended to fall within the scope of the present invention.
Example 1
A method for shortening the fermentation period of L-valine comprises the following steps:
inoculating Escherichia coli serving as a strain into a seed culture medium, controlling the fermentation temperature to 35 ℃, controlling the pH value to 6.9, controlling the tank pressure to 0.03MPa, and when the OD 610nm value is 10, ending the culture to obtain Escherichia coli seed liquid;
The seed culture medium takes water as a solvent and also comprises the following components in concentration: 10g/L of yeast powder, 25g/L of glucose, 0.5g/L of magnesium sulfate, 4g/L of dipotassium hydrogen phosphate, 4g/L of potassium dihydrogen phosphate and 4g/L of ammonium citrate.
Inoculating seed liquid into the fermentation culture medium according to the inoculum size of 13% by volume ratio for fermentation culture. Controlling the temperature to 35 ℃, the pH value to 6.9, the pressure to 0.03MPa, the ventilation quantity to 0.5m of air/m of the fermentation liquor/min, simultaneously feeding 60% (w/v) glucose solution into the fermentation liquor, controlling the glucose concentration in the fermentation liquor to 0.03% (w/v) for fermentation culture, and ending the fermentation when the L-valine content in the fermentation liquor is 7.5%, wherein the fermentation period is 23 hours. The fermentation index is shown in Table 1.
The fermentation medium takes water as a solvent and contains 5g/L of yeast powder, 0.6g/L of betaine, 10g/L of glucose, 0.2g/L of magnesium sulfate, 1g/L of dipotassium hydrogen phosphate, 1g/L of monopotassium phosphate, 2g/L of ammonium citrate, 1g/L of microelement composition and 0.2g/L of rare metal composition.
Wherein, the microelement composition takes water as a solvent and contains 1g/L of sodium selenite, 0.2g/L of boric acid, 10g/L of sodium silicate, 5g/L of sodium iodide, 2g/L of D-biotin, 20g/L of nicotinamide and 4g/L of choline chloride.
Wherein, the rare metal composition takes water as a solvent and contains 0.3g/L of nickel chloride, 3g/L of cobalt chloride, 10g/L of copper chloride, 5g/L of zinc chloride, 5g/L of sodium molybdate, 0.2g/L of scandium chloride and 0.2g/L of cesium chloride.
Example 2
A method for shortening the fermentation period of L-valine comprises the following steps:
Inoculating Escherichia coli serving as a strain into a seed culture medium, controlling the fermentation temperature to be 36 ℃, controlling the pH value to be 7.0, controlling the tank pressure to be 0.04MPa, and when the OD 610nm value is 11, ending the culture to obtain Escherichia coli seed liquid;
The seed culture medium takes water as a solvent and also comprises the following components in concentration: 11g/L of yeast powder, 27g/L of glucose, 0.6g/L of magnesium sulfate, 5g/L of dipotassium hydrogen phosphate, 5g/L of potassium dihydrogen phosphate and 6g/L of ammonium citrate.
Inoculating seed liquid into the fermentation culture medium according to the inoculum size of 14% by volume ratio for fermentation culture. The temperature is controlled to be 37 ℃, the pH value is controlled to be 7.0, the pressure is 0.04MPa, the ventilation rate is 0.65 m mu m, the air/m mu m of fermentation liquor is used for fermentation/min, 63% (w/v) glucose solution is fed into the fermentation liquor, the concentration of glucose in the fermentation liquor is controlled to be 0.04% (w/v), fermentation culture is carried out, and when the content of L-valine in the fermentation liquor is 8%, the fermentation is finished, and the fermentation period is 24 hours. The fermentation index is shown in Table 1.
The fermentation medium takes water as a solvent, contains 7g/L of yeast powder, 0.7g/L of betaine, 12g/L of glucose, 0.4g/L of magnesium sulfate, 1.5g/L of dipotassium hydrogen phosphate, 1.5g/L of monopotassium phosphate and 3g/L of ammonium citrate, 1.5g/L of microelement composition and 0.3g/L of rare metal composition.
Wherein, the microelement composition takes water as a solvent and contains 1.5g/L of sodium selenite, 0.3g/L of boric acid, 11g/L of sodium silicate, 6g/L of sodium iodide, 3g/L of D-biotin, 22g/L of nicotinamide and 5g/L of choline chloride.
Wherein, the rare metal composition takes water as a solvent and contains 0.4g/L of nickel chloride, 4g/L of cobalt chloride, 11g/L of copper chloride, 6g/L of zinc chloride, 6g/L of sodium molybdate, 0.3g/L of scandium chloride and 0.3g/L of cesium chloride.
Example 3
A method for shortening the fermentation period of L-valine comprises the following steps:
Inoculating Escherichia coli serving as a strain into a seed culture medium, controlling the fermentation temperature to 37 ℃, controlling the pH value to 7.0, controlling the tank pressure to 0.05MPa, and when the OD 610nm value is 12, ending the culture to obtain Escherichia coli seed liquid;
The seed culture medium takes water as a solvent and also comprises the following components in concentration: 12g/L of yeast powder, 30g/L of glucose, 0.8g/L of magnesium sulfate, 6g/L of dipotassium hydrogen phosphate, 6g/L of monopotassium hydrogen phosphate and 7g/L of ammonium citrate.
Inoculating seed liquid into the fermentation culture medium according to the inoculum size of 16% by volume ratio for fermentation culture. Controlling the temperature to 38 ℃, the pH value to 7.1, the pressure to 0.05MPa, the ventilation quantity to 0.8m of air/m of the fermentation liquor/min, simultaneously feeding 65% (w/v) glucose solution into the fermentation liquor, controlling the glucose concentration in the fermentation liquor to 0.05% (w/v) for fermentation culture, and ending the fermentation when the L-valine content in the fermentation liquor is 8.5%, wherein the fermentation period is 23 hours. The fermentation index is shown in Table 1.
The fermentation medium takes water as a solvent, and contains 8g/L of yeast powder, 0.8g/L of betaine, 15g/L of glucose, 0.5g/L of magnesium sulfate, 2g/L of dipotassium hydrogen phosphate, 2g/L of monopotassium phosphate and 4g/L of ammonium citrate, 2g/L of microelement composition and 0.5g/L of rare metal composition.
Wherein, the microelement composition takes water as a solvent and contains 3g/L of sodium selenite, 0.4g/L of boric acid, 12g/L of sodium silicate, 8g/L of sodium iodide, 5g/L of D-biotin, 25g/L of nicotinamide and 6g/L of choline chloride.
Wherein, the rare metal composition takes water as a solvent and contains 0.5g/L of nickel chloride, 6g/L of cobalt chloride, 15g/L of copper chloride, 8g/L of zinc chloride, 7g/L of sodium molybdate, 0.5g/L of scandium chloride and 0.4g/L of cesium chloride.
For the methods of examples 1-3, the growth of the cells OD 610nm was examined for different fermentation periods:
L-valine was produced by fermentation culture in the manner described in examples 1 to 3, sampling and detecting the cell OD 610nm every 2 hours, detecting the absorbance at a wavelength of 610nm with a spectrophotometer, and controlling the absorbance at 0.2 to 0.8 by diluting by different factors, and the OD was calculated as absorbance multiplied by the dilution factor.
As shown in FIG. 1 (the present invention only shows the growth of the cell OD 610nm in the different fermentation periods in example 2 because of the large number of pictures), the cell OD 610nm gradually increases with the increase of the culture time in the initial stage of fermentation, the cell OD 610nm reaches 11 when the period is about 8 hours, the cell starts to grow in the logarithmic phase, and the cell OD 610nm basically reaches a stable value when the fermentation is 14 hours.
The results show that the method of the invention can shorten the growth arrest phase of the thalli OD 610nm, and enable the thalli to enter the logarithmic phase rapidly, thereby shortening the fermentation period.
For the methods of examples 1-3, the L-valine content was examined for various fermentation periods:
l-valine was produced by fermentation culture in the same manner as in examples 1 to 3, and the content of L-valine in the fermentation broth was measured by sampling every 4 hours by paper chromatography.
The sample is diluted according to the content of L-valine in the fermentation broth, and the sample is spotted on a filter paper of 30 x 25cm by a spotting needle, wherein the distance between the spotting lines is 1.5cm from the bottom end of the filter paper, the distance is 2.0cm, the spotting amount is 1 mu L, and the spot concentration is 5.0% of standard substance. The filter paper with the sample is placed in a chromatography cylinder with a developing agent added in advance, the filter paper is developed by an upward method at 25 ℃, and the filter paper is taken out and dried after the front edge of the developing agent is moved to the position of 1cm above the filter paper. Spreading the air-dried filter paper after layer folding, spraying a color developing agent, heating and developing in an oven, and drying at 105 ℃ for 5min until purple amino acid spots appear on the filter paper. Purplish red spots on the filter paper were cut off, soaked in 5mL of eluent for 40min (to cover the port of the test tube, prevent ethanol evaporation), and absorbance was measured at 506 nm. And (3) taking an absorbance value obtained by a standard substance as a standard curve, checking the content of the measured diluted sample on the standard curve, and multiplying the content by the dilution times to obtain the content of L-valine in the fermentation liquid sample. Wherein the developing agent is n-butanol, glacial acetic acid and distilled water, and the volume ratio of the n-butanol, the glacial acetic acid and the distilled water is 36:9:15; the color-developing agent is ninhydrin acetone solution with the mass concentration of 0.5%; the eluent is a penta-hydrated copper sulfate solution with the mass concentration of 0.2% and ethanol with the volume concentration of 75%, and the volume ratio of the penta-hydrated copper sulfate solution to the ethanol is 2:38.
As shown in FIG. 2 (the invention only shows the results of measuring the L-valine content of different fermentation periods in example 2 because of more pictures), and with reference to FIG. 1, the bacterial cells are in a lag phase for 6 hours before fermentation, the L-valine synthesis rate is slower, and when the bacterial cells OD 610nm reaches 10, the bacterial cells enter a rapid growth phase, and the L-valine synthesis rate is gradually increased.
The results show that the method provided by the invention can rapidly accumulate and produce L-valine and improve the fermentation production intensity.
Comparative example 1
L-valine was prepared by fermentation in the same manner as in example 1 except that neither composition was added to the fermentation medium, and the other was unchanged. The results are shown in FIGS. 3-4.
As can be seen from FIGS. 3 to 4, in the case where neither composition was added, the cell growth arrest phase was long, the log phase growth was started substantially only about 16 hours in the period, the stable value was reached substantially at 28 hours in the period, and L-valine was slowly produced in acid and the fermentation strength was low. The fermentation index is shown in Table 1.
Comparative example 2
L-valine was produced by fermentation in the same manner as in example 2 except that the trace element composition was added to the fermentation medium, and the rare metal composition was not added thereto, and the other was unchanged. The fermentation index is shown in Table 1.
Comparative example 3
L-valine was produced by fermentation in the same manner as in example 3 except that the rare metal composition alone was added to the fermentation medium, and the trace element composition was not added thereto, and the other was unchanged. The fermentation index is shown in Table 1.
Table 1 fermentation index data in each of examples and comparative examples 1 to 3
| Fermentation period h | Acid content% | Conversion% | |
| Example 1 | 23 | 7.5 | 51.85 |
| Example 2 | 24 | 8 | 52.08 |
| Example 3 | 23 | 8.5 | 51.91 |
| Comparative example 1 | 36 | 7.53 | 40.34 |
| Comparative example 2 | 30 | 8.14 | 41.85 |
| Comparative example 3 | 34 | 8.47 | 45.85 |
The acid content refers to the content of L-valine in the fermentation broth at the end of fermentation;
Conversion refers to the acid production amount (L-valine production amount)/glucose amount×100%; wherein the glucose amount is the total amount of glucose utilized in the fermentation process; the acid yield refers to the acid content x the volume of the material discharged, which refers to the volume of all the material in the fermenter at the end of the fermentation.
In addition, since there is an error in the experimental process, although comparative example 1 is based on the method of example 1, the control of the L-valine content in the fermentation broth may have small up-and-down fluctuations at the end of fermentation; comparative example 2 and comparative example 3 are also the same problems.
As can be seen from the data in table 1:
According to the invention, the three embodiments of adding the trace element composition and the rare metal composition have the advantages of shortened fermentation period of 23-24h and higher conversion rate of more than 51%. The method has the advantages that the trace element composition and the rare metal composition are added simultaneously, so that the rapid growth and propagation of thalli can be promoted, the fermentation period is shortened, the metabolic enzyme activity of thalli can be improved, and the fermentation conversion rate of L-valine can be improved.
Comparative example 1 (neither composition was added), the fermentation period was longer, the cell growth was slow, the lag phase was longer, and L-valine accumulation was slower; the fermentation conversion rate of L-valine is low and is only 40.34 percent.
Comparative example 2 (only trace element composition is added, no rare metal composition is added), the fermentation period is shortened more than comparative example 1, and the addition of trace element composition can promote the rapid growth and propagation of thalli and shorten the fermentation period; but has less effect on improving the metabolic enzyme activity of the thalli.
Comparative example 3 (only rare metal composition is added, no trace element composition is added), the L-valine conversion rate is improved more than comparative example 1, and the rare metal composition can improve the bacterial metabolism enzyme activity and the L-valine fermentation conversion rate; but has less effect of promoting the rapid growth and propagation of the thalli.
Verification example 1
L-valine was prepared by fermentation in the same manner as in example 1 except that sodium selenite was not added to the trace element composition, and the other was unchanged.
Verification example 2
L-valine was prepared by fermentation in the same manner as in example 1 except that boric acid was not added to the trace element composition, and the other was unchanged.
Verification example 3
L-valine was prepared by fermentation in the same manner as in example 1 except that sodium silicate was not added to the trace element composition, and the other was unchanged.
Verification example 4
L-valine was prepared by fermentation in the same manner as in example 1 except that sodium iodide was not added to the trace element composition, and the other was unchanged.
Verification example 5
L-valine was prepared by fermentation in the same manner as in example 1 except that D-biotin was not added to the trace element composition, and the other was unchanged.
Verification example 6
L-valine was prepared by fermentation in the same manner as in example 1 except that nicotinamide was not added to the trace element composition, and the other was unchanged.
Verification example 7
L-valine was prepared by fermentation in the same manner as in example 1 except that choline chloride was not added to the trace element composition, and the other was unchanged.
Verification example 8
L-valine was prepared by fermentation in the same manner as in example 1 except that nickel chloride was not added to the rare earth metal composition, and the other was unchanged.
Verification example 9
L-valine was prepared by fermentation in the same manner as in example 1 except that cobalt chloride was not added to the rare earth metal composition, and the other was unchanged.
Verification example 10
L-valine was prepared by fermentation in the same manner as in example 1 except that copper chloride was not added to the rare earth metal composition, and the other was unchanged.
Verification example 11
L-valine was prepared by fermentation in the same manner as in example 1 except that zinc chloride was not added to the rare earth metal composition, and the other was unchanged.
Verification example 12
L-valine was produced by fermentation in the same manner as in example 1 except that sodium molybdate was not added to the rare earth metal composition, and the other was not changed.
Verification example 13
L-valine was produced by fermentation in the same manner as in example 1 except that scandium chloride was not added to the rare earth metal composition, and the other was not changed.
Verification example 14
L-valine was prepared by fermentation in the same manner as in example 1 except that cesium chloride was not added to the rare earth metal composition, and the other was unchanged.
TABLE 2 fermentation index data for each of the verification examples
| Fermentation period h | Acid content% | Conversion% | |
| Verification example 1 | 26 | 7.56 | 50.45 |
| Verification example 2 | 28 | 7.51 | 50.39 |
| Verification example 3 | 27 | 7.53 | 51.05 |
| Verification example 4 | 27 | 7.47 | 50.98 |
| Verification example 5 | 27 | 7.55 | 50.30 |
| Verification example 6 | 26 | 7.49 | 50.58 |
| Verification example 7 | 26 | 7.45 | 50.67 |
| Verification example 8 | 24 | 7.56 | 48.62 |
| Verification example 9 | 23 | 7.51 | 47.98 |
| Verification example 10 | 25 | 7.47 | 48.35 |
| Verification example 11 | 24 | 7.46 | 47.84 |
| Verification example 12 | 24 | 7.54 | 48.25 |
| Verification example 13 | 23 | 7.48 | 48.11 |
| Verification example 14 | 24 | 7.55 | 47.88 |
Because of errors in the experimental process, although the test examples 1 to 14 were based on the method of example 1, the control of the L-valine content in the fermentation broth at the end of the fermentation may have small fluctuations.
The fermentation index data of each test example are shown in table 2, and it can be seen that:
in the verification examples 1 to 7, when a certain element is absent in the trace element composition, the fermentation period is prolonged by 2 to 3 hours compared with the example, which shows that the trace element composition is the best trace element composition suitable for producing L-valine by fermentation, and the absence of any one has an influence on the fermentation period;
In the verification examples 8 to 14, when a certain element is absent from the rare metal composition, the fermentation period is substantially the same as that of the examples, but the fermentation conversion rate is lower than that of the examples, which indicates that the rare metal composition is the best rare metal composition suitable for the fermentation production of L-valine, and the absence of any one has an influence on the fermentation acid production and conversion rate;
Therefore, the microelement composition and the rare metal composition in the embodiment are the optimal proportion and the optimal composition suitable for fermentation production of L-valine, the fermentation period can be shortened to the greatest extent, the fermentation acid production can be improved, and the lack of any one has a great influence on fermentation.
Comparative example 4
L-valine was produced by fermentation in the same manner as in example 3 except that the ventilation was 0.2 m g/m.mu.m.g/min during fermentation, and the other was unchanged.
Comparative example 5
L-valine was produced by fermentation in the same manner as in example 3 except that the ventilation amount was 1.1 m air/mW of fermentation broth/min during fermentation, and the other was unchanged.
Comparative example 6
L-valine was produced by fermentation in the same manner as in example 3 except that the glucose concentration in the fermentation broth was controlled to be 0.01% (w/v) during the fermentation, and the other was not changed.
Comparative example 7
L-valine was produced by fermentation in the same manner as in example 3 except that the glucose concentration in the fermentation broth was controlled to be 0.1% (w/v) during the fermentation, and the other was not changed.
TABLE 3 fermentation index data in comparative examples 4-7
| Fermentation period h | Acid content% | Conversion% | |
| Example 1 | 23 | 7.5 | 51.85 |
| Example 2 | 24 | 8 | 52.08 |
| Example 3 | 23 | 8.5 | 51.91 |
| Comparative example 4 | 26 | 8.49 | 42.69 |
| Comparative example 5 | 24 | 8.53 | 51.96 |
| Comparative example 6 | 25 | 8.55 | 44.16 |
| Comparative example 7 | 25 | 8.47 | 45.77 |
Because of errors in the experimental process, although comparative examples 4 to 7 were based on the method of example 3, there was a possibility that the control of L-valine content in the fermentation broth could be slightly fluctuating up and down at the end of fermentation.
The normal fermentation ventilation rate of L-valine is generally 0.2-0.4m, and air/m is the fermentation broth/min, but as the fermentation medium is improved, the growth and propagation speed of thalli is higher, and metabolism is vigorous, so that the oxygen amount needs to be increased, and as can be seen from the data of comparative example 4, when the ventilation rate is lower than 0.5 m m, the fermentation broth/min is the fermentation broth/min, the fermentation of L-valine is influenced, and the conversion rate is reduced; as can be seen from the data of comparative example 5, the conversion was not affected when the aeration level was higher than 0.8 m m of air/m of broth/min.
As the microelement composition and the rare metal composition are added, the bacteria are well metabolized, the enzyme activity is improved, the low-sugar fermentation is more favorable for accumulating and producing L-valine, and the synthesis of other mixed acids such as organic acid is reduced. As can be seen from comparative example 6, the glucose concentration is too low, the fermentation is incomplete, and the conversion rate is relatively low; as can be seen from comparative example 7, the glucose concentration was too high, but the conversion was low, indicating that the metabolite was doped with a lot of hetero acids.
The foregoing description of the preferred embodiments of the invention is not intended to be limiting, but rather is intended to cover all modifications, equivalents, alternatives, and improvements that fall within the spirit and scope of the invention.
Claims (8)
1. A method for shortening the fermentation period of L-valine, characterized by: taking escherichia coli as a strain, adding 1-2g/L of microelement composition and 0.2-0.5g/L of rare metal composition into a fermentation culture medium for fermentation culture, wherein:
The microelement composition takes water as a solvent and also comprises the following components in concentration: 1-3g/L of sodium selenite, 0.2-0.4g/L of boric acid, 10-12g/L of sodium silicate, 5-8g/L of sodium iodide, 2-5g/L of D-biotin, 20-25g/L of nicotinamide and 4-6g/L of choline chloride;
The rare metal composition takes water as a solvent and further comprises the following components in concentration: 0.3-0.5g/L of nickel chloride, 3-6g/L of cobalt chloride, 10-15g/L of copper chloride, 5-8g/L of zinc chloride, 5-7g/L of sodium molybdate, 0.2-0.5g/L of scandium chloride and 0.2-0.4g/L of cesium chloride;
the fermentation culture method comprises the following steps: inoculating the escherichia coli seed liquid into a fermentation culture medium, controlling the ventilation rate to be 0.5-0.8 m mu m, adding 60-65% w/v glucose solution in a flowing mode, and controlling the glucose concentration in the fermentation liquid to be 0.03-0.05% w/v for fermentation culture.
2. The method for shortening the fermentation period of L-valine according to claim 1, wherein: the fermentation medium takes water as a solvent and also comprises the following basic components in concentration: 5-8g/L of yeast powder, 0.6-0.8g/L of betaine, 10-15g/L of glucose, 0.2-0.5g/L of magnesium sulfate, 1-2g/L of dipotassium hydrogen phosphate, 1-2g/L of potassium dihydrogen phosphate and 2-4g/L of ammonium citrate.
3. The method for shortening the fermentation period of L-valine according to claim 1, wherein: inoculating the escherichia coli seed liquid into a fermentation culture medium, controlling the temperature to be 35-38 ℃, the pH value to be 6.9-7.1, and the pressure to be 0.03-0.05MPa.
4. The method for shortening the fermentation period of L-valine according to claim 1, wherein: the inoculation amount of the escherichia coli seed liquid is 13-16% of the volume of the fermentation medium.
5. The method for shortening a fermentation period of L-valine according to claim 4, wherein: the preparation method of the escherichia coli seed liquid comprises the following steps: inoculating the escherichia coli into a seed culture medium for seed culture until the OD 610nm value of the thalli is 10-12, and obtaining escherichia coli seed liquid.
6. The method for shortening a fermentation period of L-valine according to claim 5, wherein: the seed culture medium takes water as a solvent and also comprises the following components in concentration: 10-12g/L of yeast powder, 25-30g/L of glucose, 0.5-0.8g/L of magnesium sulfate, 4-6g/L of dipotassium hydrogen phosphate, 4-6g/L of potassium dihydrogen phosphate and 4-7g/L of ammonium citrate.
7. The method for shortening a fermentation period of L-valine according to claim 5, wherein: the culture temperature of the escherichia coli seed liquid is 35-37 ℃, the pH value is 6.9-7.0, and the pressure is 0.03-0.05MPa.
8. The method for shortening the fermentation period of L-valine according to claim 1, wherein: in the fermentation culture process, when the content of L-valine in the fermentation liquid is 7.5-8.5%, the fermentation is finished; at the end of fermentation, the fermentation period of L-valine is 23-24h.
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| CN1982466A (en) * | 2005-12-12 | 2007-06-20 | 上海化工研究院 | Production of stabilized isotope 15N labelled L-valine |
| CN117701459A (en) * | 2023-12-22 | 2024-03-15 | 苏州百因诺生物科技有限公司 | Escherichia coli high-density fermentation medium and fermentation process |
| CN117757861A (en) * | 2022-09-19 | 2024-03-26 | 廊坊梅花生物技术开发有限公司 | Valine production method and application |
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| CN1982466A (en) * | 2005-12-12 | 2007-06-20 | 上海化工研究院 | Production of stabilized isotope 15N labelled L-valine |
| CN117757861A (en) * | 2022-09-19 | 2024-03-26 | 廊坊梅花生物技术开发有限公司 | Valine production method and application |
| CN117701459A (en) * | 2023-12-22 | 2024-03-15 | 苏州百因诺生物科技有限公司 | Escherichia coli high-density fermentation medium and fermentation process |
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