CN114875090B - Method for producing lysine and application thereof - Google Patents
Method for producing lysine and application thereof Download PDFInfo
- Publication number
- CN114875090B CN114875090B CN202210616257.4A CN202210616257A CN114875090B CN 114875090 B CN114875090 B CN 114875090B CN 202210616257 A CN202210616257 A CN 202210616257A CN 114875090 B CN114875090 B CN 114875090B
- Authority
- CN
- China
- Prior art keywords
- fermentation
- glucose
- producing
- lysine
- threonine
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
- 239000004472 Lysine Substances 0.000 title claims abstract description 66
- KDXKERNSBIXSRK-UHFFFAOYSA-N Lysine Natural products NCCCCC(N)C(O)=O KDXKERNSBIXSRK-UHFFFAOYSA-N 0.000 title claims abstract description 64
- 238000004519 manufacturing process Methods 0.000 title claims abstract description 25
- 238000000855 fermentation Methods 0.000 claims abstract description 171
- 230000004151 fermentation Effects 0.000 claims abstract description 166
- 239000004473 Threonine Substances 0.000 claims abstract description 61
- AYFVYJQAPQTCCC-UHFFFAOYSA-N Threonine Natural products CC(O)C(N)C(O)=O AYFVYJQAPQTCCC-UHFFFAOYSA-N 0.000 claims abstract description 51
- 238000000034 method Methods 0.000 claims abstract description 26
- 241000894006 Bacteria Species 0.000 claims abstract description 19
- 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 45
- 239000008103 glucose Substances 0.000 claims description 45
- 239000002609 medium Substances 0.000 claims description 34
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 claims description 28
- QGZKDVFQNNGYKY-UHFFFAOYSA-N Ammonia Chemical compound N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 claims description 27
- 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 description 20
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 claims description 20
- 229910052921 ammonium sulfate Inorganic materials 0.000 claims description 20
- 235000011130 ammonium sulphate Nutrition 0.000 claims description 20
- 240000008042 Zea mays Species 0.000 claims description 19
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 claims description 19
- 235000002017 Zea mays subsp mays Nutrition 0.000 claims description 19
- 235000005822 corn Nutrition 0.000 claims description 19
- XKMRRTOUMJRJIA-UHFFFAOYSA-N ammonia nh3 Chemical compound N.N XKMRRTOUMJRJIA-UHFFFAOYSA-N 0.000 claims description 18
- 235000019764 Soybean Meal Nutrition 0.000 claims description 16
- 239000004455 soybean meal Substances 0.000 claims description 16
- 229910052757 nitrogen Inorganic materials 0.000 claims description 14
- 229910021529 ammonia Inorganic materials 0.000 claims description 13
- 239000001963 growth medium Substances 0.000 claims description 13
- 239000002054 inoculum Substances 0.000 claims description 13
- 235000013379 molasses Nutrition 0.000 claims description 13
- VHUUQVKOLVNVRT-UHFFFAOYSA-N Ammonium hydroxide Chemical compound [NH4+].[OH-] VHUUQVKOLVNVRT-UHFFFAOYSA-N 0.000 claims description 12
- 235000011114 ammonium hydroxide Nutrition 0.000 claims description 12
- 239000000413 hydrolysate Substances 0.000 claims description 12
- 229960002685 biotin Drugs 0.000 claims description 10
- 235000020958 biotin Nutrition 0.000 claims description 10
- 239000011616 biotin Substances 0.000 claims description 10
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 claims description 9
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 claims description 9
- 229910052799 carbon Inorganic materials 0.000 claims description 9
- 108010009736 Protein Hydrolysates Proteins 0.000 claims description 4
- 238000005273 aeration Methods 0.000 claims description 4
- 241000588724 Escherichia coli Species 0.000 claims description 3
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 claims description 3
- 229930006000 Sucrose Natural products 0.000 claims description 3
- 238000007599 discharging Methods 0.000 claims description 3
- 238000004321 preservation Methods 0.000 claims description 3
- 239000005720 sucrose Substances 0.000 claims description 3
- 230000001105 regulatory effect Effects 0.000 claims description 2
- 239000001166 ammonium sulphate Substances 0.000 claims 1
- 239000000758 substrate Substances 0.000 abstract description 7
- 238000003912 environmental pollution Methods 0.000 abstract description 4
- 230000008569 process Effects 0.000 abstract description 3
- AYFVYJQAPQTCCC-GBXIJSLDSA-N L-threonine Chemical compound C[C@@H](O)[C@H](N)C(O)=O AYFVYJQAPQTCCC-GBXIJSLDSA-N 0.000 description 65
- 229960002898 threonine Drugs 0.000 description 55
- KDXKERNSBIXSRK-YFKPBYRVSA-N L-lysine Chemical compound NCCCC[C@H](N)C(O)=O KDXKERNSBIXSRK-YFKPBYRVSA-N 0.000 description 54
- 235000018977 lysine Nutrition 0.000 description 52
- 239000007788 liquid Substances 0.000 description 15
- 239000002253 acid Substances 0.000 description 10
- 238000012360 testing method Methods 0.000 description 9
- 230000001276 controlling effect Effects 0.000 description 7
- 238000006555 catalytic reaction Methods 0.000 description 6
- 229940063746 oxygen 20 % Drugs 0.000 description 6
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 6
- 238000006243 chemical reaction Methods 0.000 description 5
- 239000000463 material Substances 0.000 description 5
- 238000011218 seed culture Methods 0.000 description 5
- 229940024606 amino acid Drugs 0.000 description 4
- 235000001014 amino acid Nutrition 0.000 description 4
- 150000001413 amino acids Chemical class 0.000 description 4
- 108090000790 Enzymes Proteins 0.000 description 3
- 102000004190 Enzymes Human genes 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- 238000011081 inoculation Methods 0.000 description 3
- 238000012986 modification Methods 0.000 description 3
- 230000004048 modification Effects 0.000 description 3
- 230000037361 pathway Effects 0.000 description 3
- 244000144977 poultry Species 0.000 description 3
- 239000008223 sterile water Substances 0.000 description 3
- 238000009423 ventilation Methods 0.000 description 3
- QGZKDVFQNNGYKY-UHFFFAOYSA-O Ammonium Chemical compound [NH4+] QGZKDVFQNNGYKY-UHFFFAOYSA-O 0.000 description 2
- CURLTUGMZLYLDI-UHFFFAOYSA-N Carbon dioxide Chemical compound O=C=O CURLTUGMZLYLDI-UHFFFAOYSA-N 0.000 description 2
- 108010064711 Homoserine dehydrogenase Proteins 0.000 description 2
- CKLJMWTZIZZHCS-REOHCLBHSA-N L-aspartic acid Chemical compound OC(=O)[C@@H](N)CC(O)=O CKLJMWTZIZZHCS-REOHCLBHSA-N 0.000 description 2
- 241001052560 Thallis Species 0.000 description 2
- 239000003674 animal food additive Substances 0.000 description 2
- 235000003704 aspartic acid Nutrition 0.000 description 2
- OQFSQFPPLPISGP-UHFFFAOYSA-N beta-carboxyaspartic acid Natural products OC(=O)C(N)C(C(O)=O)C(O)=O OQFSQFPPLPISGP-UHFFFAOYSA-N 0.000 description 2
- 238000012258 culturing Methods 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 238000009472 formulation Methods 0.000 description 2
- 244000144972 livestock Species 0.000 description 2
- 230000004060 metabolic process Effects 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 230000029219 regulation of pH Effects 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- UKAUYVFTDYCKQA-UHFFFAOYSA-N -2-Amino-4-hydroxybutanoic acid Natural products OC(=O)C(N)CCO UKAUYVFTDYCKQA-UHFFFAOYSA-N 0.000 description 1
- 108010055400 Aspartate kinase Proteins 0.000 description 1
- 101710088194 Dehydrogenase Proteins 0.000 description 1
- 102100021758 E3 ubiquitin-protein transferase MAEA Human genes 0.000 description 1
- 102000010911 Enzyme Precursors Human genes 0.000 description 1
- 108010062466 Enzyme Precursors Proteins 0.000 description 1
- 235000019766 L-Lysine Nutrition 0.000 description 1
- UKAUYVFTDYCKQA-VKHMYHEASA-N L-homoserine Chemical compound OC(=O)[C@@H](N)CCO UKAUYVFTDYCKQA-VKHMYHEASA-N 0.000 description 1
- AGPKZVBTJJNPAG-WHFBIAKZSA-N L-isoleucine Chemical compound CC[C@H](C)[C@H](N)C(O)=O AGPKZVBTJJNPAG-WHFBIAKZSA-N 0.000 description 1
- FFEARJCKVFRZRR-BYPYZUCNSA-N L-methionine Chemical compound CSCC[C@H](N)C(O)=O FFEARJCKVFRZRR-BYPYZUCNSA-N 0.000 description 1
- 108010043075 L-threonine 3-dehydrogenase Proteins 0.000 description 1
- 102100026808 Mitochondrial import inner membrane translocase subunit Tim8 A Human genes 0.000 description 1
- GNISQJGXJIDKDJ-YFKPBYRVSA-N O-succinyl-L-homoserine Chemical compound OC(=O)[C@@H](N)CCOC(=O)CCC(O)=O GNISQJGXJIDKDJ-YFKPBYRVSA-N 0.000 description 1
- 102100033451 Thyroid hormone receptor beta Human genes 0.000 description 1
- 230000004913 activation Effects 0.000 description 1
- GZCGUPFRVQAUEE-SLPGGIOYSA-N aldehydo-D-glucose Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@@H](O)C=O GZCGUPFRVQAUEE-SLPGGIOYSA-N 0.000 description 1
- 230000004075 alteration Effects 0.000 description 1
- 238000005576 amination reaction Methods 0.000 description 1
- 125000003277 amino group Chemical group 0.000 description 1
- 230000036528 appetite Effects 0.000 description 1
- 235000019789 appetite Nutrition 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 239000012620 biological material Substances 0.000 description 1
- 229910002092 carbon dioxide Inorganic materials 0.000 description 1
- 239000001569 carbon dioxide Substances 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 208000019425 cirrhosis of liver Diseases 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 230000002950 deficient Effects 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 238000004134 energy conservation Methods 0.000 description 1
- 239000003623 enhancer Substances 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- HHLFWLYXYJOTON-UHFFFAOYSA-N glyoxylic acid Chemical compound OC(=O)C=O HHLFWLYXYJOTON-UHFFFAOYSA-N 0.000 description 1
- 238000004128 high performance liquid chromatography Methods 0.000 description 1
- 108010071598 homoserine kinase Proteins 0.000 description 1
- 238000009776 industrial production Methods 0.000 description 1
- 229960000310 isoleucine Drugs 0.000 description 1
- AGPKZVBTJJNPAG-UHFFFAOYSA-N isoleucine Natural products CCC(C)C(N)C(O)=O AGPKZVBTJJNPAG-UHFFFAOYSA-N 0.000 description 1
- 210000005229 liver cell Anatomy 0.000 description 1
- 230000003908 liver function Effects 0.000 description 1
- 230000037353 metabolic pathway Effects 0.000 description 1
- 229930182817 methionine Natural products 0.000 description 1
- 230000000813 microbial effect Effects 0.000 description 1
- 230000002906 microbiologic effect Effects 0.000 description 1
- 238000009629 microbiological culture Methods 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 238000012544 monitoring process Methods 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 230000016087 ovulation Effects 0.000 description 1
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 230000008929 regeneration Effects 0.000 description 1
- 238000011069 regeneration method Methods 0.000 description 1
- 230000001954 sterilising effect Effects 0.000 description 1
- 230000004936 stimulating effect Effects 0.000 description 1
- 239000013589 supplement Substances 0.000 description 1
- 230000004102 tricarboxylic acid cycle Effects 0.000 description 1
Classifications
-
- 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
- C12P13/04—Alpha- or beta- amino acids
- C12P13/08—Lysine; Diaminopimelic acid; Threonine; Valine
-
- 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
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
- C12R2001/185—Escherichia
- C12R2001/19—Escherichia coli
Landscapes
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Organic Chemistry (AREA)
- Health & Medical Sciences (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Genetics & Genomics (AREA)
- Biotechnology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- Microbiology (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- Biomedical Technology (AREA)
- Virology (AREA)
- Tropical Medicine & Parasitology (AREA)
- Medicinal Chemistry (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
The invention relates to the technical field of biological fermentation, and particularly discloses a method for producing lysine and application thereof. According to the method for producing lysine, fermentation bacteria for producing lysine and fermentation bacteria for producing threonine are subjected to co-fermentation culture in the same fermentation medium. The method can not only improve the substrate utilization rate, but also save labor and energy, simplify process equipment and reduce environmental pollution, and provides a novel method for producing lysine.
Description
Technical Field
The invention relates to the technical field of biological fermentation, in particular to a method for producing lysine and application thereof.
Background
Lysine is a basic amino acid of formula C 6 H 14 N 2 O 2 Molecular weight 146.19, which contains two amino groups in its molecule. Lysine has wide application as food enhancer and medicine as liver cell regeneration agent, and has obvious curative effect on improving liver function, treating liver cirrhosis, stimulating appetite and improving nutrient condition. As a feed additive, a small amount of lysine is added into the feed of livestock and poultry, and the feed additive has remarkable effects on daily gain, feed conversion ratio, ovulation amount of poultry and the like of the poultry and the livestock.
Metabolic pathways for producing lysine from glucose include an EMP pathway, an HMP pathway, a TCA cycle, a glyoxylate cycle, a carbon dioxide fixation reaction, an aminoacylacetic acid amination reaction to produce aspartic acid, aspartic acid to produce cycloaspartic acid- β -semialdehyde under the catalysis of aspartokinase and aspartic acid- β -semialdehyde dehydrogenase, and then split into two pathways, on the one hand, lysine is produced under the catalysis of a series of enzymes such as DDP synthase, on the other hand, homoserine is produced under the catalysis of homoserine dehydrogenase, on the other hand, methionine is produced under the catalysis of a series of enzymes such as succinyl homoserine synthase, on the other hand, threonine is produced under the catalysis of enzymes such as homoserine kinase, and isoleucine is produced under the catalysis of threonine dehydrogenase, etc.
The lysine-producing microorganism can improve lysine production by a auxotroph such as threonine-deficient strain. Threonine can be added into the formula in the lysine fermentation process to meet the growth requirement of lysine-producing bacteria. Threonine is also an amino acid produced by microbial fermentation. The production of threonine or lysine by fermentation requires separate fermentation equipment, raw materials and auxiliary materials, and energy sources, which are not beneficial to saving cost and avoiding environmental pollution, so that further research on the production and fermentation of lysine is necessary.
Disclosure of Invention
The invention aims to provide a lysine production method with high substrate utilization rate, labor and energy conservation, simplified process equipment and little environmental pollution.
The technical scheme of the invention is as follows:
a method for producing lysine, which comprises co-fermenting and culturing a fermentation bacterium producing lysine and a fermentation bacterium producing threonine in the same fermentation medium.
In the method of the invention, the zymogen for producing lysine is MHZ-0914, and the preservation number of MHZ-0914 is CGMCC No.22648;
the fermentation bacteria for producing threonine is MHZ-0216-5.
The invention provides a novel method for producing alkaline amino acid-lysine, which is to put a strain for producing lysine into a culture medium for fermentation culture, and put a strain for producing threonine into the culture medium for mixed culture with lysine bacteria. Threonine produced by the threonine producing strain is used as a nitrogen source for producing lysine by the lysine producing strain, and is used for the growth and metabolism of lysine producing bacteria to complete fermentation and produce lysine, so that the production of the lysine nitrogen source and the production of lysine are simultaneously carried out in the same substrate, the threonine does not need to be produced before the lysine is produced, the fermentation process and the fermentation substrate are integrally saved, the substrate utilization rate is improved, and the industrial production cost is greatly saved.
In the method of the present invention, the ratio of the inoculum size of the lysine-producing fermentation cylinder to that of the threonine-producing fermentation cylinder is 1: (0.04-0.16), preferably 1:0.08.
In the method, during the co-fermentation culture, the concentration of the initial fermentation glucose is 19-21g/L, the fermentation temperature is 36-38 ℃, the fermentation pressure is controlled to be 0.07-0.09MPa, and the ventilation rate is 0.9-1.1vvm; glucose and ammonium sulfate are continuously added in the fermentation process, the concentration of residual sugar in the fermentation process is controlled to be 0-1g/L, and ammonia Nitrogen (NH) is fermented 4 + ) The concentration is 0.8-1.2g/L, ammonia water is used for pH regulation and control, the fermentation pH value is 6.7-7.1, and the fermentation time is 35-37h; and discharging when the volume of the culture solution in the fermentation tank reaches 68-72% of the total volume of the fermentation tank, wherein the discharge volume is 4.8-5.2% of the volume of the culture medium.
According to the invention, lysine-producing bacteria and threonine-producing bacteria are used as research objects, the amount of threonine required in mixed culture is considered from the amount of threonine required in the independent lysine fermentation process, and the growth and metabolism of the lysine-producing bacteria are not influenced (the mixed fermentation reaches the pure fermentation lysine fermentation index level) by controlling the specific inoculation proportion and the ammonium ion concentration in the fermentation process, and the growth of the threonine-producing bacteria is controlled specifically, so that the threonine yield is controlled, the lysine production effect in the mixed fermentation is good, and the substrate utilization rate is high.
In the method of the present invention, the carbon source of the fermentation medium includes glucose and sucrose, and the nitrogen source includes ammonium sulfate and ammonia water.
In the method of the present invention, the fermentation medium comprises:
glucose 19-21g/L, H 3 PO 4 1.2-1.7g/L，KCl 0.4-0.6g/L，MgSO 4 0.6-0.8g/L, 9-11g/L molasses, 58-62g/L corn steep liquor, 29-31g/L soybean meal hydrolysate, mnSO 4 0.0018-0.0022g/L，FeSO 4 0.0018-0.0022g/L，VB 1 95-105 mug/L, 190-210 mug/L biotin; glucose and ammonium sulfate were fed in.
Preferably, the fermentation medium comprises: glucose 20g/L, H 3 PO 4 1.5g/L，KCl 0.5g/L，MgSO 4 0.7g/L，10g/L molasses, 60g/L corn steep liquor, 30g/L soybean meal hydrolysate and MnSO 4 0.002g/L，FeSO 4 0.002g/L，VB 1 100. Mu.g/L, biotin 200. Mu.g/L.
In the method of the present invention, the seed medium of the lysine-producing fermentation broth comprises: glucose 39-41g/L KH 2 PO 4 1.4-1.6g/L，MgSO 4 1.4-1.6g/L, molasses 11-13g/L, corn steep liquor 24-26g/L, ammonium sulfate 11-13g/L, mnSO 4 0.0015-0.0025g/L，FeSO 4 0.0015-0.0025g/L。
In the method of the present invention, the seed medium of threonine-producing fermentation broth includes: glucose 18-22g/L, corn steep liquor 10-20g/L, soybean meal hydrolysate 3-7g/L and MgSO 4 ·7H 2 O 0.4-0.6g/L，KH 2 PO 4 1.8-2.2g/L。
The invention also provides an application of the method in reducing the cost of producing lysine.
The invention has the advantages that:
the invention carries out fermentation culture on two different zymophytes under the same condition, thereby improving the substrate utilization rate, saving labor and energy, simplifying process equipment, reducing environmental pollution and providing a novel method for producing lysine.
Drawings
FIG. 1 shows the results of lysine fermentation OD and acid production in the 50L fermentation tank of MHZ-0914 in example 1 of the present invention.
FIG. 2 shows the results of threonine fermentation OD and acid production in the 50L fermenter of MHZ-0216-5 in example 2 of the present invention.
FIG. 3 shows the results of fermentation OD and acid production in 50L fermentors with different inoculum sizes of MHZ-0216-5 in example 3 according to the present invention.
FIG. 4 shows the results of fermentation OD and acid production in 50L fermentors under the control of ammonia nitrogen concentration in different fermentation processes of MHZ-0216-5 in example 4 of the present invention.
FIG. 5 shows the results of the co-fermentation OD and acid production in the MHZ-0914 and MHZ-0216-5 50L fermentors of example 5 of the present invention.
Detailed Description
Preferred embodiments of the present invention will be described in detail below with reference to examples. It is to be understood that the following examples are given for illustrative purposes only and are not intended to limit the scope of the present invention. Various modifications and alterations of this invention may be made by those skilled in the art without departing from the spirit and scope of this invention.
The experimental methods used in the following examples are conventional methods unless otherwise specified. Materials, reagents and the like used in the examples described below are commercially available unless otherwise specified.
In the present invention, the OD value is the result of the test at a wavelength of 600nm unless otherwise specified.
In the embodiment of the invention, the experimental method is as follows:
(1) Pure culture fermentation for producing lysine and threonine is respectively carried out;
(2) Inoculating threonine bacteria into lysine fermentation culture medium according to different inoculum sizes for pure culture fermentation;
(3) The lysine bacteria and threonine bacteria are inoculated into a lysine fermentation medium according to a certain proportion, and mixed fermentation is carried out.
(1) Inoculating L-lysine and L-threonine producing strains in a bacteria-preserving tube stored at-80 ℃ on a slant culture medium for activation;
(2) inoculating thalli on the slant culture medium into a first-stage seed shake flask for culture;
(3) the first-level seed liquid is inoculated into a 10L fermentation tank for secondary seed culture;
(4) and (3) inoculating the secondary seed liquid into a fermentation medium, and fermenting and culturing in a 50L fermentation tank.
And (3) condition control:
(1) ammonia concentration control: the ammonia concentration as a nitrogen source required for amino acid production cannot be in a low state in the medium, which would otherwise lead to a decrease in the productivity of basic amino acids. Ammonia is fed in, and simultaneously, ammonium sulfate is fed in, so that the ammonia concentration in the fermentation liquor is kept at 0.8-1.2g/L, preferably 1g/L.
(2) Sugar concentration and pH control: the proportional relationship between the two is obtained by examining the consumption conditions of acid production, sugar and ammonia of thalli in the fermentation process. According to the method, pH feedback signals are used as control conditions, zero sugar is used for control in fermentation liquor, so that the pH feedback system can realize the supplement of sugar while ammonia is fed into a fermentation tank, a Kjeldahl nitrogen analyzer is used for monitoring the content of free ammonia in the fermentation process, and recording is carried out every 6 hours.
Example 1
The present example uses a lysine-producing strain: MHZ-0914, fermentation alone.
Biomaterial MHZ-0914, class designation: escherichia coli deposited in China general microbiological culture Collection center (CMCC) at 2021, month 6 and day 1: the Beijing city, the Korean region, the North Chen West way No. 1, the national academy of sciences of China, the microbiological institute, is postal code 100101, and the preservation number is CGMCC No.22648.
Seed medium (g/L): glucose 40, KH 2 PO 4 1.5，MgSO 4 1.5, molasses 12, corn steep liquor 25, ammonium sulfate 12, mnSO 4 0.002，FeSO 4 0.002。
Fermentation medium: glucose 20g/L, H 3 PO 4 1.5g/L，KCl 0.5g/L，MgSO 4 0.7g/L, 10g/L molasses, 60g/L corn steep liquor, 30g/L soybean meal hydrolysate and MnSO 4 0.002g/L，FeSO 4 0.002g/L，VB 1 100. Mu.g/L, biotin 200. Mu.g/L. 600g/L glucose, 500g/L ammonium sulfate and total nitrogen are fed in: threonine 4g/L. Sterilizing at 121deg.C for 20min.
The seed tanks are respectively connected with the lysine production strains, when the OD value reaches 0.8, the seed tanks are connected with a fermentation tank for fermentation, the fermentation inoculation ratio is 20%, the initial fermentation glucose concentration is 20g/L, the fermentation temperature is 36-38 ℃, the fermentation pressure is controlled to be 0.08MPa, the ventilation rate is 1vvm, the culture is 36 hours, the fermentation tanks are continuously fed with fermentable sugar which is high-concentration glucose solution with the concentration of 600g/L, the fed ammonium sulfate concentration is 500g/L, the fed total nitrogen is threonine, the total nitrogen addition amount accounts for 22% of the volume of the glucose solution addition amount, the residual sugar in the fermentation process is 1g/L, and the fermentation ammonia nitrogen (inorganic free ammonia NH) 4 + ) The concentration is 0.8-1.2g/L, ammonia water is used for pH regulation and control, the fermentation pH value is 6.7-7.1, and when the volume of the culture solution in the fermentation tank reaches 70% of the volume of the fermentation tank, the culture solution is putThe material and the discharge volume are 5% of the volume of the culture medium, and the fermentation time is 36h. During fermentation, the acid and free ammonia are measured during fermentation.
As a result of fermentation, referring to FIG. 1, it was found that the fermentation was carried out for 36 hours and that lysine was present at 190g/L. The total sugar content of the fermentation is 10050g, the conversion rate is 65.2%, the theoretical acid is 6552g, the highest OD value is 34, the threonine consumption is 14.5g, and the ammonia nitrogen in the fermentation process is controlled to be 0.8-1.2g/L.
Example 2
The present example employs threonine producing strains: MHZ-0216-5, and fermenting separately.
The L-threonine-producing strain in this example is Escherichia coli MHZ-0216-5 (see Chinese patent CN113846132A for strain MHZ-0216-5).
Seed culture medium: 20g/L glucose, 15g/L corn steep liquor, 5g/L soybean meal hydrolysate and MgSO 4 ·7H 2 O0.5g/L，KH 2 PO 4 2g/L。
Fermentation medium: 40g/L glucose, 7g/L corn steep liquor, 7g/L soybean meal hydrolysate and MgSO 4 ·7H 2 O0.5g/L，KH 2 PO 4 2g/L。
The sterilized L-threonine seed medium was added to a sterile seed tank 10L, water was added to adjust the initial volume of the seed medium to 6L, and 200mL of seed solution was inoculated. Culture conditions: aeration rate 0.8vvm, rotation speed 300rpm, culture pH7.0, temperature 37 ℃ and dissolved oxygen 20%. The culture was stopped when the seeds grew until the OD value was 10.
Inoculating sterilized L-threonine fermentation medium into a sterile fermentation tank 50L, and adding sterile water to a volume of 15L; and 3L of seed liquid in the seed tank is taken and connected into the fermentation tank. Fermentation process control conditions: 0.5-0.8vvm,300-700rpm, tank pressure 0.07MPa, pH7.0, temperature 37 ℃, dissolved oxygen 20%, fermentation period 36h.
When the residual sugar content in the fermentation liquid is 0.1g/L, starting to feed a carbon source, controlling pH to 7.0 by taking ammonia water as a pH regulator, preparing a 50% glucose solution as the fed-batch carbon source, controlling pH to 7.0 by taking 25% ammonia water as the fed-batch nitrogen source in the fermentation process, and controlling the concentration of glucose in the fermentation liquid to be about 1g/L in the fermentation process.
The glucose content was determined using an SBA biosensor, the free ammonium content was determined using a Kjeldahl nitrogen determination instrument, and the L-threonine content was determined using HPLC.
As a result of fermentation, as shown in FIG. 2, it was revealed that the fermentation was carried out for 36 hours at an L-threonine concentration of 110g/L and a conversion rate of 54%, and that the highest OD value was 30 and the threonine yield was 2540g.
Example 3
The present example employs threonine producing strains: MHZ-0216-5, and fermenting separately. Using the lysine fermentation formulation of example 1, inoculum sizes were 1L,0.5L,0.25L, and 0.125L, respectively.
Seed culture medium: 20g/L glucose, 15g/L corn steep liquor, 5g/L soybean meal hydrolysate and MgSO 4 ·7H 2 O0.5g/L，KH 2 PO 4 2g/L。
Fermentation medium: glucose 20g/L, H 3 PO 4 1.5g/L，KCl 0.5g/L，MgSO 4 0.7g/L, 10g/L molasses, 60g/L corn steep liquor, 30g/L soybean meal hydrolysate and MnSO 4 0.002g/L，FeSO 4 0.002g/L，VB 1 100. Mu.g/L, biotin 200. Mu.g/L. 600g/L of glucose and 500g/L of ammonium sulfate are fed in, and threonine is not fed in.
The sterilized L-threonine seed medium was added to a sterile seed tank 10L, water was added to adjust the initial volume of the seed medium to 6L, and 200mL of seed solution was inoculated. Culture conditions: aeration rate 0.8vvm, rotation speed 300rpm, culture pH7.0, temperature 37 ℃ and dissolved oxygen 20%. The culture was stopped when the seeds grew until the OD value was 10.
The sterilized L-threonine fermentation medium is inoculated into a sterile fermentation tank 50L, sterile water is added to a volume of 15L, and seed liquid is inoculated. Fermentation process control conditions: 0.5-0.8vvm,300-700rpm, tank pressure 0.07MPa, pH7.0, temperature 37 ℃, dissolved oxygen 20%, fermentation period 36h.
When the residual sugar content in the fermentation liquor is 0.1g/L, starting to feed carbon source, using nitrogen source ammonia water as pH regulator to control pH7.0, preparing 600g/L glucose solution as fed carbon source, using 25% ammonia water to control pH7.0, controlling glucose concentration in the fermentation liquor to about 1g/L in fermentation process, using 500g/L ammonium sulfate to control fermentation ammonia nitrogen (inorganic free ammonia NH) 4 + ) The concentration is 0.8-1.2g/L.
The fermentation results are shown in FIG. 3, wherein Thr-1 and OD-1 are respectively the test results with the inoculum size of 1L, thr-2 and OD-2 are respectively the test results with the inoculum size of 0.5L, thr-3 and OD-3 are respectively the test results with the inoculum size of 0.25L, thr-4 and OD-4 are respectively the test results with the inoculum size of 0.125L, and the fermentation time is 36h, the threonine concentration of 1L is 7.2g/L, the fermentation liquid volume is 18.88L, and the yield is 136g; inoculating 0.5L threonine with concentration of 5g/L, fermenting liquid with volume of 16L, and yield of 80g; inoculating 0.25L threonine with concentration of 4.4g/L, fermenting liquid volume of 15.9L, and yield of 70g; the threonine concentration of 0.125L of inoculation is 3.1g/L, the fermentation liquid volume is 15.48L, and the yield is 48g.
Example 4
The present example employs threonine producing strains: MHZ-0216-5, and fermenting separately. Using the lysine fermentation formulation of example 1, the inoculum size was 0.25L.
Seed culture medium: 20g/L glucose, 15g/L corn steep liquor, 5g/L soybean meal hydrolysate and MgSO 4 ·7H 2 O0.5g/L，KH 2 PO 4 2g/L。
Fermentation medium: glucose 20g/L, H 3 PO 4 1.5g/L，KCl 0.5g/L，MgSO 4 0.7g/L, 10g/L molasses, 60g/L corn steep liquor, 30g/L soybean meal hydrolysate and MnSO 4 0.002g/L，FeSO 4 0.002g/L，VB 1 100. Mu.g/L, biotin 200. Mu.g/L. 600g/L of fed glucose, no threonine and ammonium sulfate (with the concentration of 500 g/L) are fed in, so that ammonia nitrogen (inorganic free ammonia NH) in the fermentation process is controlled 4 + ) The concentration is 0.5g/L,1.0g/L,1.5g/L and 2.0g/L respectively.
The sterilized L-threonine seed medium was added to a sterile seed tank 10L, water was added to adjust the initial volume of the seed medium to 6L, and 200mL of seed solution was inoculated. Culture conditions: aeration rate 0.8vvm, rotation speed 300rpm, culture pH7.0, temperature 37 ℃ and dissolved oxygen 20%. The culture was stopped when the seeds grew until the OD value was 10.
The sterilized L-threonine fermentation medium is inoculated into a sterile fermentation tank 50L, sterile water is added to a volume of 15L, and seed liquid is inoculated. Fermentation process control conditions: 0.5-0.8vvm,300-700rpm, tank pressure 0.07MPa, pH7.0, temperature 37 ℃, dissolved oxygen 20%, fermentation period 36h.
When the residual sugar content in the fermentation liquor is 0.1g/L, starting to feed carbon source, using nitrogen source ammonia water as pH regulator to control pH7.0, preparing 600g/L glucose solution as fed carbon source, using 25% ammonia water to control pH7.0, controlling glucose concentration in the fermentation liquor to about 1g/L, fermenting ammonia nitrogen (inorganic free ammonia NH) 4 + ) The concentration is controlled by the ammonium sulfate fed-batch and is respectively 0.5g/L,1.0g/L,1.5g/L and 2.0g/L.
Referring to FIG. 4, wherein Thr-1 and OD-1 are respectively the test results of ammonia nitrogen concentration control of 0.5g/L, thr-2 and OD-2 are respectively the test results of ammonia nitrogen concentration control of 1g/L, thr-3 and OD-3 are respectively the test results of ammonia nitrogen concentration control of 1.5g/L, thr-4 and OD-4 are respectively the test results of ammonia nitrogen concentration control of 2g/L, and from the results, 36h of fermentation, threonine concentration of 0.5g/L, threonine concentration of 18.0L of fermentation liquid volume and yield of 72g are obtained; the concentration of ammonia nitrogen is controlled to be 1g/L, the concentration of threonine is controlled to be 2.7g/L, the volume of fermentation liquid is 17.3L, and the yield is 46.7g; ammonia nitrogen concentration is controlled to be 1.5g/L, threonine concentration is controlled to be 1.8g/L, fermentation liquid volume is controlled to be 16.6L, and yield is controlled to be 30g; the concentration of ammonia nitrogen is controlled to be 2.0g/L, the concentration of threonine is controlled to be 1g/L, the volume of fermentation liquid is 15.5L, and the yield is 15.5g.
Example 5
Seed culture was performed on the lysine-producing strain MHZ-0914 and the threonine-producing strain MHZ-0216-5 as described in examples 1 and 2, respectively, and then seed solutions of the above two strains were inoculated into 15L of the lysine fermentation medium in example 1, the lysine inoculum size was 20%, and the threonine inoculum size was 1.6%.
Specifically, when threonine seeds and lysine seeds reach the OD of the lower tank, the threonine seeds and the lysine seeds are simultaneously inoculated into a lysine fermentation culture medium, the initial fermentation glucose concentration is 20g/L, the fermentation temperature is 36-38 ℃, the fermentation pressure is controlled to be 0.08MPa, the ventilation rate is 1vvm, the culture is carried out for 36 hours, and 600g/L of high-concentration glucose solution and 500g/L of ammonium sulfate are continuously fed into the fermentation tank. The residual sugar in the fermentation process is zero sugar, the ammonia nitrogen concentration of the fermentation is 0.8-1.2g/L, the pH value of the fermentation is 6.7-7.1 by regulating and controlling the pH value of the fermentation by 25% ammonia water, the fermentation culture is carried out, when the volume of the culture solution in the fermentation tank reaches 70% of the volume of the fermentation tank, the material is discharged, the material discharging volume is 5% of the volume of the culture medium, and the fermentation time is 36h.
As shown in the fermentation result in FIG. 5, the fermentation time is 36h, the lysine content is 192g/L, the total sugar content is 10105g, the conversion rate is 65.3%, the theoretical acid content is 6598g, the threonine content is 0g, and the ammonia nitrogen content in the fermentation process is controlled to be 0.8-1.2g/L.
While the invention has been described in detail in the foregoing general description and with reference to specific embodiments thereof, it will be apparent to one skilled in the art that modifications and improvements can be made thereto. Accordingly, such modifications or improvements may be made without departing from the spirit of the invention and are intended to be within the scope of the invention as claimed.
Claims (14)
1. A method for producing lysine is characterized in that fermentation bacteria for producing lysine and fermentation bacteria for producing threonine are subjected to co-fermentation culture in the same fermentation medium;
the fermentation bacteria for producing lysine is MHZ-0914, and the preservation number of the MHZ-0914 is CGMCC No.22648; the fermentation bacteria for producing threonine are escherichia coli MHZ-0216-5.
2. The method according to claim 1, wherein the ratio of the inoculum size of the lysine-producing fermentation cylinder to the threonine-producing fermentation cylinder is 1: (0.04-0.16).
3. The method according to claim 2, wherein the ratio of the inoculum size of the lysine-producing fermentation cylinder to the threonine-producing fermentation cylinder is 1:0.08.
4. A method according to any one of claims 1 to 3, wherein the initial fermentation glucose concentration is 19 to 21g/L, the fermentation temperature is 36 to 38 ℃, the fermentation pressure is controlled to be 0.07 to 0.09MPa, and the aeration rate is 0.9 to 1.1vvm when the co-fermentation culture is performed; glucose and ammonium sulfate are continuously added in the fermentation process, the concentration of residual sugar in the fermentation process is controlled to be 0-1g/L, the concentration of ammonia nitrogen in the fermentation process is controlled to be 0.8-1.2g/L, pH is regulated and controlled by ammonia water, the fermentation pH value is 6.7-7.1, and the fermentation time is 35-37h; and discharging when the volume of the culture solution in the fermentation tank reaches 68-72% of the total volume of the fermentation tank, wherein the discharge volume is 4.8-5.2% of the volume of the culture medium.
5. A method according to any one of claims 1-3, characterized in that the carbon source of the fermentation medium comprises glucose and sucrose and the nitrogen source comprises ammonium sulphate and ammonia.
6. The method of claim 4, wherein the carbon source of the fermentation medium comprises glucose and sucrose and the nitrogen source comprises ammonium sulfate and aqueous ammonia.
7. The method according to any one of claims 1-3, 6, wherein the fermentation medium comprises:
glucose 19-21g/L, H 3 PO 4 1.2-1.7g/L，KCl 0.4-0.6g/L，MgSO 4 0.6-0.8g/L, 9-11g/L molasses, 58-62g/L corn steep liquor, 29-31g/L soybean meal hydrolysate, mnSO 4 0.0018-0.0022g/L，FeSO 4 0.0018-0.0022g/L，VB 1 95-105 mug/L, 190-210 mug/L biotin; glucose and ammonium sulfate were fed in.
8. The method of claim 4, wherein the fermentation medium comprises:
glucose 19-21g/L, H 3 PO 4 1.2-1.7g/L，KCl 0.4-0.6g/L，MgSO 4 0.6-0.8g/L, 9-11g/L molasses, 58-62g/L corn steep liquor, 29-31g/L soybean meal hydrolysate, mnSO 4 0.0018-0.0022g/L，FeSO 4 0.0018-0.0022g/L，VB 1 95-105 mug/L, 190-210 mug/L biotin; glucose and ammonium sulfate were fed in.
9. The method of claim 5, wherein the fermentation medium comprises:
glucose 19-21g/L, H 3 PO 4 1.2-1.7g/L，KCl 0.4-0.6g/L，MgSO 4 0.6-0.8g/L, 9-11g/L molasses, 58-62g/L corn steep liquor, 29-31g/L soybean meal hydrolysate, mnSO 4 0.0018-0.0022g/L，FeSO 4 0.0018-0.0022g/L，VB 1 95-105 mug/L, 190-210 mug/L biotin; glucose and ammonium sulfate were fed in.
10. The method of claim 7, wherein the fermentation medium comprises: glucose 20g/L, H 3 PO 4 1.5g/L，KCl 0.5g/L，MgSO 4 0.7g/L, 10g/L molasses, 60g/L corn steep liquor, 30g/L soybean meal hydrolysate and MnSO 4 0.002g/L，FeSO 4 0.002g/L，VB 1 100. Mu.g/L, biotin 200. Mu.g/L.
11. The method according to claim 8 or 9, wherein the fermentation medium comprises: glucose 20g/L, H 3 PO 4 1.5g/L，KCl 0.5g/L，MgSO 4 0.7g/L, 10g/L molasses, 60g/L corn steep liquor, 30g/L soybean meal hydrolysate and MnSO 4 0.002g/L，FeSO 4 0.002g/L，VB 1 100. Mu.g/L, biotin 200. Mu.g/L.
12. A method according to any one of claims 1-3, wherein the seed medium of the lysine producing fermentation broth comprises: glucose 39-41g/L KH 2 PO 4 1.4-1.6g/L，MgSO 4 1.4-1.6g/L, molasses 11-13g/L, corn steep liquor 24-26g/L, ammonium sulfate 11-13g/L, mnSO 4 0.0015-0.0025g/L，FeSO 4 0.0015-0.0025g/L。
13. A method according to any one of claims 1-3, characterized in that the seed medium of threonine-producing fermentation broth comprises: glucose 18-22g/L, corn steep liquor 10-20g/L, soybean meal hydrolysate 3-7g/L and MgSO 4 ·7H 2 O 0.4-0.6g/L，KH 2 PO 4 1.8-2.2g/L。
14. Use of the method of any one of claims 1-13 for reducing the cost of producing lysine.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210616257.4A CN114875090B (en) | 2022-05-31 | 2022-05-31 | Method for producing lysine and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210616257.4A CN114875090B (en) | 2022-05-31 | 2022-05-31 | Method for producing lysine and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN114875090A CN114875090A (en) | 2022-08-09 |
| CN114875090B true CN114875090B (en) | 2024-03-26 |
Family
ID=82680301
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202210616257.4A Active CN114875090B (en) | 2022-05-31 | 2022-05-31 | Method for producing lysine and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN114875090B (en) |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| GB1118719A (en) * | 1966-03-14 | 1968-07-03 | Kyowa Hakko Kogyo Kk | Process for producing l-lysine by fermentation |
| RU2347807C1 (en) * | 2007-12-13 | 2009-02-27 | Общество с ограниченной ответственностью "БИОРЕАКТОР" | Escherichia coli-lysine producer strain, method of making feed additive, containing this strain, composition, obtained using this method and method of monogastric animals and birds |
| CN113846132A (en) * | 2020-06-28 | 2021-12-28 | 廊坊梅花生物技术开发有限公司 | Construction of threonine producing strain and method for producing threonine |
-
2022
- 2022-05-31 CN CN202210616257.4A patent/CN114875090B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| GB1118719A (en) * | 1966-03-14 | 1968-07-03 | Kyowa Hakko Kogyo Kk | Process for producing l-lysine by fermentation |
| RU2347807C1 (en) * | 2007-12-13 | 2009-02-27 | Общество с ограниченной ответственностью "БИОРЕАКТОР" | Escherichia coli-lysine producer strain, method of making feed additive, containing this strain, composition, obtained using this method and method of monogastric animals and birds |
| CN113846132A (en) * | 2020-06-28 | 2021-12-28 | 廊坊梅花生物技术开发有限公司 | Construction of threonine producing strain and method for producing threonine |
Non-Patent Citations (1)
| Title |
|---|
| A Programmable Escherichia coli Consortium via Tunable Symbiosis;Alissa Kerner等;PLoS ONE;第7卷(第3期);doi:10.1371/ journal.pone.0034032，第1-10页 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN114875090A (en) | 2022-08-09 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN1072721C (en) | Process for producing L-leucine | |
| CN114875090B (en) | Method for producing lysine and application thereof | |
| JP3074781B2 (en) | Production method of L-lysine by fermentation method | |
| CN110885774A (en) | Method for optimizing glutamic acid fermentation | |
| CN112852896A (en) | Fermentation production method of L-arginine | |
| CA1301690C (en) | PROCESS FOR THE FERMENTATIVE PREPARATION OF L-AMINO ACIDS FROM .alpha.-KETO CARBOXYLIC ACIDS | |
| US20050032159A1 (en) | Method for low-temperature culture of microorganism | |
| CN110923275A (en) | Fermentation and extraction process of glutamic acid | |
| CN112029683A (en) | Glucose control process for improving L-isoleucine yield | |
| CN112662609B (en) | Fermentation medium for increasing yield of beta-alanine and application method | |
| CN1183474A (en) | Process for producing L-lysine by fermentation | |
| CN116287046A (en) | Amino acid fermentation medium and method for producing lysine by semi-continuous fermentation | |
| CN116042753B (en) | Preparation method of corn steep liquor mycoprotein composite hydrolysate and preparation method of L-isoleucine | |
| CN114606276B (en) | Method for improving fermentation yield of L-threonine | |
| EP2287324A2 (en) | Fermentation processes with low concentrations of carbon- and nitrogen-containing nutrients | |
| CN117431277A (en) | Method for producing L-amino acid by fermentation | |
| CN118222647A (en) | Method for improving lysine fermentation yield and reducing by-product acetic acid | |
| CN112080533B (en) | Full-nutrition fed-batch fermentation control process for improving yield of L-isoleucine | |
| CN101321860A (en) | Fermentation process for preparing L-lysine | |
| CN116656755A (en) | Culture medium sterilization method and application thereof in L-amino acid fermentation production | |
| CN116606795A (en) | Method for improving amino acid fermentation acid production | |
| CN116218926A (en) | Method for producing acidic amino acid and application thereof | |
| CN117402919A (en) | Method for producing amino acid of aspartic acid family by fermentation and application thereof | |
| CN106520744A (en) | Method for improving nitrile hydratase activity of bacillus subtilis | |
| CN117903983A (en) | Corynebacterium glutamicum mutant strain, microbial inoculum thereof and application of corynebacterium glutamicum mutant strain in L-arginine fermentation |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |