CN116287046A - Amino acid fermentation medium and method for producing lysine by semi-continuous fermentation - Google Patents
Amino acid fermentation medium and method for producing lysine by semi-continuous fermentation Download PDFInfo
- Publication number
- CN116287046A CN116287046A CN202210726965.3A CN202210726965A CN116287046A CN 116287046 A CN116287046 A CN 116287046A CN 202210726965 A CN202210726965 A CN 202210726965A CN 116287046 A CN116287046 A CN 116287046A
- Authority
- CN
- China
- Prior art keywords
- fermentation
- lysine
- tank
- producing
- continuous
- 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.)
- Pending
Links
- 238000000855 fermentation Methods 0.000 title claims abstract description 144
- 230000004151 fermentation Effects 0.000 title claims abstract description 144
- 239000004472 Lysine Substances 0.000 title claims abstract description 45
- KDXKERNSBIXSRK-UHFFFAOYSA-N Lysine Natural products NCCCCC(N)C(O)=O KDXKERNSBIXSRK-UHFFFAOYSA-N 0.000 title claims abstract description 45
- 238000004519 manufacturing process Methods 0.000 title claims abstract description 35
- 150000001413 amino acids Chemical class 0.000 title claims abstract description 22
- 239000002609 medium Substances 0.000 claims abstract description 21
- PVNIIMVLHYAWGP-UHFFFAOYSA-N Niacin Chemical compound OC(=O)C1=CC=CN=C1 PVNIIMVLHYAWGP-UHFFFAOYSA-N 0.000 claims abstract description 19
- 239000001963 growth medium Substances 0.000 claims abstract description 18
- 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
- 229960003512 nicotinic acid Drugs 0.000 claims abstract description 10
- 235000001968 nicotinic acid Nutrition 0.000 claims abstract description 10
- 239000011664 nicotinic acid Substances 0.000 claims abstract description 10
- 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 7
- 229930003451 Vitamin B1 Natural products 0.000 claims abstract description 7
- 240000008042 Zea mays Species 0.000 claims abstract description 7
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 claims abstract description 7
- 235000002017 Zea mays subsp mays Nutrition 0.000 claims abstract description 7
- 235000020958 biotin Nutrition 0.000 claims abstract description 7
- 239000011616 biotin Substances 0.000 claims abstract description 7
- 229960002685 biotin Drugs 0.000 claims abstract description 7
- 235000005822 corn Nutrition 0.000 claims abstract description 7
- 239000008103 glucose Substances 0.000 claims abstract description 7
- 235000013379 molasses Nutrition 0.000 claims abstract description 7
- 229960003495 thiamine Drugs 0.000 claims abstract description 7
- DPJRMOMPQZCRJU-UHFFFAOYSA-M thiamine hydrochloride Chemical compound Cl.[Cl-].CC1=C(CCO)SC=[N+]1CC1=CN=C(C)N=C1N DPJRMOMPQZCRJU-UHFFFAOYSA-M 0.000 claims abstract description 7
- 239000011691 vitamin B1 Substances 0.000 claims abstract description 7
- 235000010374 vitamin B1 Nutrition 0.000 claims abstract description 7
- 235000019764 Soybean Meal Nutrition 0.000 claims abstract description 6
- 239000004455 soybean meal Substances 0.000 claims abstract description 6
- 108010009736 Protein Hydrolysates Proteins 0.000 claims abstract description 5
- 235000001014 amino acid Nutrition 0.000 claims description 20
- 235000000346 sugar Nutrition 0.000 claims description 10
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 claims description 7
- 229910052921 ammonium sulfate Inorganic materials 0.000 claims description 7
- 235000011130 ammonium sulphate Nutrition 0.000 claims description 7
- 235000015097 nutrients Nutrition 0.000 claims description 7
- 238000004321 preservation Methods 0.000 claims description 6
- FFEARJCKVFRZRR-BYPYZUCNSA-N L-methionine Chemical compound CSCC[C@H](N)C(O)=O FFEARJCKVFRZRR-BYPYZUCNSA-N 0.000 claims description 4
- AYFVYJQAPQTCCC-GBXIJSLDSA-N L-threonine Chemical compound C[C@@H](O)[C@H](N)C(O)=O AYFVYJQAPQTCCC-GBXIJSLDSA-N 0.000 claims description 4
- AYFVYJQAPQTCCC-UHFFFAOYSA-N Threonine Natural products CC(O)C(N)C(O)=O AYFVYJQAPQTCCC-UHFFFAOYSA-N 0.000 claims description 4
- 239000004473 Threonine Substances 0.000 claims description 4
- 235000006109 methionine Nutrition 0.000 claims description 4
- 229930182817 methionine Natural products 0.000 claims description 4
- 235000008521 threonine Nutrition 0.000 claims description 4
- 238000007599 discharging Methods 0.000 abstract description 16
- 241001052560 Thallis Species 0.000 abstract description 10
- 238000000034 method Methods 0.000 abstract description 6
- 230000002503 metabolic effect Effects 0.000 abstract description 4
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 abstract description 2
- 230000006978 adaptation Effects 0.000 abstract description 2
- 229910052799 carbon Inorganic materials 0.000 abstract description 2
- 238000009776 industrial production Methods 0.000 abstract description 2
- 238000005457 optimization Methods 0.000 abstract description 2
- 239000007788 liquid Substances 0.000 abstract 1
- 239000002253 acid Substances 0.000 description 20
- 239000000243 solution Substances 0.000 description 11
- 238000006243 chemical reaction Methods 0.000 description 10
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 8
- 230000006872 improvement Effects 0.000 description 7
- 241001465754 Metazoa Species 0.000 description 5
- 239000000463 material Substances 0.000 description 5
- 235000016709 nutrition Nutrition 0.000 description 5
- 230000035764 nutrition Effects 0.000 description 5
- 244000005700 microbiome Species 0.000 description 4
- 229910052757 nitrogen Inorganic materials 0.000 description 4
- 230000004060 metabolic process Effects 0.000 description 3
- QGZKDVFQNNGYKY-UHFFFAOYSA-N Ammonia Chemical compound N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 description 2
- XKMRRTOUMJRJIA-UHFFFAOYSA-N ammonia nh3 Chemical compound N.N XKMRRTOUMJRJIA-UHFFFAOYSA-N 0.000 description 2
- 230000001276 controlling effect Effects 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 239000000413 hydrolysate Substances 0.000 description 2
- 238000009629 microbiological culture Methods 0.000 description 2
- 239000011259 mixed solution Substances 0.000 description 2
- 239000000047 product Substances 0.000 description 2
- 230000001105 regulatory effect Effects 0.000 description 2
- 238000009423 ventilation Methods 0.000 description 2
- 241000894006 Bacteria Species 0.000 description 1
- 241001061906 Caragana Species 0.000 description 1
- 241000186226 Corynebacterium glutamicum Species 0.000 description 1
- 101710088194 Dehydrogenase Proteins 0.000 description 1
- 241000588724 Escherichia coli Species 0.000 description 1
- BAWFJGJZGIEFAR-NNYOXOHSSA-N NAD zwitterion Chemical compound NC(=O)C1=CC=C[N+]([C@H]2[C@@H]([C@H](O)[C@@H](COP([O-])(=O)OP(O)(=O)OC[C@@H]3[C@H]([C@@H](O)[C@@H](O3)N3C4=NC=NC(N)=C4N=C3)O)O2)O)=C1 BAWFJGJZGIEFAR-NNYOXOHSSA-N 0.000 description 1
- XJLXINKUBYWONI-NNYOXOHSSA-N NADP zwitterion Chemical compound NC(=O)C1=CC=C[N+]([C@H]2[C@@H]([C@H](O)[C@@H](COP([O-])(=O)OP(O)(=O)OC[C@@H]3[C@H]([C@@H](OP(O)(O)=O)[C@@H](O3)N3C4=NC=NC(N)=C4N=C3)O)O2)O)=C1 XJLXINKUBYWONI-NNYOXOHSSA-N 0.000 description 1
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 description 1
- LCTONWCANYUPML-UHFFFAOYSA-M Pyruvate Chemical compound CC(=O)C([O-])=O LCTONWCANYUPML-UHFFFAOYSA-M 0.000 description 1
- 238000005273 aeration Methods 0.000 description 1
- 230000037354 amino acid metabolism Effects 0.000 description 1
- 239000003674 animal food additive Substances 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 239000005515 coenzyme Substances 0.000 description 1
- 230000000052 comparative effect Effects 0.000 description 1
- AWADHHRPTLLUKK-UHFFFAOYSA-N diazanium sulfuric acid sulfate Chemical compound [NH4+].[NH4+].OS(O)(=O)=O.[O-]S([O-])(=O)=O AWADHHRPTLLUKK-UHFFFAOYSA-N 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 238000006911 enzymatic reaction Methods 0.000 description 1
- 230000006545 glycolytic metabolism Effects 0.000 description 1
- 239000001257 hydrogen Substances 0.000 description 1
- 229910052739 hydrogen Inorganic materials 0.000 description 1
- 125000004435 hydrogen atom Chemical class [H]* 0.000 description 1
- 230000007062 hydrolysis Effects 0.000 description 1
- 238000006460 hydrolysis reaction Methods 0.000 description 1
- 230000037353 metabolic pathway Effects 0.000 description 1
- 230000000813 microbial effect Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 229950006238 nadide Drugs 0.000 description 1
- 230000003204 osmotic effect Effects 0.000 description 1
- 230000003647 oxidation Effects 0.000 description 1
- 238000007254 oxidation reaction Methods 0.000 description 1
- 150000002972 pentoses Chemical class 0.000 description 1
- 235000011007 phosphoric acid Nutrition 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 235000018102 proteins Nutrition 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 238000006479 redox reaction Methods 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 230000001502 supplementing effect Effects 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
Images
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
- 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
- C12N1/38—Chemical stimulation of growth or activity by addition of chemical compounds which are not essential growth factors; Stimulation of growth by removal of a chemical compound
-
- 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
- C12R2001/15—Corynebacterium
-
- 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
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P60/00—Technologies relating to agriculture, livestock or agroalimentary industries
- Y02P60/80—Food processing, e.g. use of renewable energies or variable speed drives in handling, conveying or stacking
- Y02P60/87—Re-use of by-products of food processing for fodder production
Landscapes
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Life Sciences & Earth Sciences (AREA)
- Organic Chemistry (AREA)
- Health & Medical Sciences (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Genetics & Genomics (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Biotechnology (AREA)
- General Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- Microbiology (AREA)
- General Chemical & Material Sciences (AREA)
- Medicinal Chemistry (AREA)
- Tropical Medicine & Parasitology (AREA)
- Virology (AREA)
- Biomedical Technology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
The invention provides an amino acid fermentation medium and a method for producing lysine by semi-continuous fermentation, belonging to the technical field of fermentation engineering. The fermentation medium comprises: glucose, H 3 PO 4 ，KCl，MgSO 4 Molasses, corn steep liquor, soybean meal hydrolysate, nicotinic acid and MnSO 4 ，FeSO 4 Vitamin B1 and biotin. The invention improves the metabolic capability of the thalli through the optimization of the culture medium, also maximally utilizes the thalli in the culture medium, and avoids the attenuation of the thalli in the culture medium due to lack of carbon source after discharging in conventional productionThe artificial consumption of fermentation capacity, and the adaptation period of continuous fermentation liquid without thallus reproduction shortens the production period and improves the production efficiency. In addition, the production conditions have no larger variation, the operation is simple, and the method is suitable for large-scale industrial production.
Description
Technical Field
The invention relates to the technical field of fermentation engineering, in particular to an amino acid fermentation medium and a method for producing lysine by semi-continuous fermentation.
Background
Lysine (Lysine) is one of many amino acids essential to humans and animals and is commonly found in biological proteins. Conventionally, the production methods of lysine include hydrolysis, chemical synthesis and enzymatic methods, and with the rise of microbial fermentation and the development of biotechnology, new fermentation technologies gradually replace the conventional production methods. The lysine product is mainly used as feed additive for animals, and has important effects of improving metabolic condition in animals, promoting growth and development of animals, improving nutrition utilization rate of animals on feed, etc.
Nicotinic acid is an important participant in oxidation-reduction reaction, and coenzyme I and coenzyme II mainly serve as the coenzyme of dehydrogenase and play a role of hydrogen transfer body in biological oxidation, have important roles in microorganisms, participate in glycolytic metabolism, pyruvate metabolism, pentose metabolic pathway and the like, and are closely related to amino acid metabolism.
At present, lysine production in China is mostly carried out in a fed-batch fermentation mode. According to the fermentation tank, corresponding materials are added and added according to a certain volume ratio, and meanwhile, continuous discharging is carried out for producing lysine by fermentation, so that fermentation liquor accumulated by discharging cannot play a role in the mode along with the fermentation, and the bacteria with vigorous metabolism are decayed due to lack of nutrition, so that the acid production efficiency of strains is influenced due to the fact that acid produced in the discharging acid and the fermentation tank are greatly different, single-batch fermentation conversion rate is low, acid production is low, lysine productivity is limited finally, and production benefits are reduced.
Disclosure of Invention
The invention aims to provide an amino acid fermentation medium and a method for producing lysine by semi-continuous fermentation, which are used for improving fermentation strength by utilizing the promotion effect of nicotinic acid in a fermentation metabolism way, and simultaneously discharging materials in a fermentation process and throwing the materials into a secondary fermentation tank for secondary fermentation, so that the discharged thalli are utilized, the acid yield of a single fermentation tank is further improved, and the lysine productivity is improved on the basis of not limiting the fermentation conversion rate.
The technical scheme of the invention is realized as follows:
the present invention provides an amino acid fermentation medium comprising: 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, 100-120mg/L nicotinic acid, mnSO 4 1.8-2.2mg/L，FeSO 4 1.8-2.2mg/L, vitamin B1 95-105 mug/L, biotin 190-210 mug/L.
The invention further provides a method for producing lysine by semi-continuous fermentation, which comprises the steps of utilizing a lysine production strain, performing expansion culture by a seed tank, inoculating the strain into the amino acid fermentation culture medium of claim 1 for primary fermentation, periodically discharging part of fermentation liquor when the volume of the culture medium in the primary fermentation tank reaches a certain proportion of the volume of the fermentation tank, supplementing the amino acid fermentation culture medium in the primary fermentation tank, discharging the fermentation liquor into a secondary fermentation tank, continuously feeding nutrients, continuously performing secondary fermentation, discharging the fermentation liquor together by the secondary fermentation tank and the primary fermentation tank, and entering a treatment flow of the fermentation liquor to obtain lysine.
As a further improvement of the invention, the lysine producing strain is MHZ-0914 and/or MHZ-0913-3, the preservation number of MHZ-0914 is CGMCC No.22648, and the preservation number of MHZ-0913-3 is CGMCC No.11942.
As a further improvement of the present invention, the lysine-producing strain is MHZ-0914.
As a further improvement of the present invention, the fed-batch nutrients include sugar, ammonium sulfate, threonine and methionine.
As a further improvement of the invention, the culture solution in the fermentation tank is discharged when the volume of the culture solution in the fermentation tank is 65-75% of the volume of the fermentation tank, and the discharge volume is 5-10% of the volume of the fermentation solution.
As a further development of the invention, the secondary fermenter and the primary fermenter are provided with 1 or more.
As a further improvement of the invention, the secondary fermentation tank and the primary fermentation tank are both provided with a feeding system.
As a further improvement of the invention, the secondary fermentation tank is free of culture medium before discharging, and fermentation is continued by feeding nutrient substances after discharging.
The invention has the following beneficial effects: the nicotinic acid is added into the amino acid fermentation medium, so that the metabolic capability of thalli can be promoted, the fermentation consumption level is improved, the fermentation strength is further improved, and finally, the aim of increasing the acid yield of a single batch can be fulfilled.
Meanwhile, the adopted semi-continuous fermentation method is characterized in that fresh culture medium is continuously supplemented, the fermentation tank is used for circularly discharging, so that the osmotic pressure in a fermentation tank system is reduced, the mass transfer environment in the tank is improved, and meanwhile, the discharging part is supplemented with the fresh culture medium for fermentation after being moved into the next-stage fermentation tank, so that the maximum utilization of thalli is realized.
According to the invention, the metabolic capacity of the thalli is improved through the optimization of the culture medium, the thalli in the culture medium is utilized to the greatest extent, the artificial consumption of the fermentation capacity caused by the decay of the thalli in the culture medium due to the lack of a carbon source after the discharging in the conventional production is avoided, meanwhile, the adaptation period of continuous discharging fermentation liquor without the thalli reproduction is shortened, the production period is shortened, and the production efficiency is improved. In addition, the production conditions have no larger variation, the operation is simple, and the method is suitable for large-scale industrial production.
Drawings
In order to more clearly illustrate the embodiments of the invention or the technical solutions of the prior art, the drawings which are used in the description of the embodiments or the prior art will be briefly described, it being obvious that the drawings in the description below are only some embodiments of the invention, and that other drawings can be obtained according to these drawings without inventive faculty for a person skilled in the art.
FIG. 1 is a schematic illustration of the process flow of the present invention.
Detailed Description
The following description of the technical solutions in the embodiments of the present invention will be clear and complete, and it is obvious that the described embodiments are only some embodiments of the present invention, but not all embodiments. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention.
The lysine producing strain is MHZ-0914 and/or MHZ-0913-3, the preservation number of the lysine producing strain MHZ-0914 is CGMCC No.22648, and the strain is preserved in China general microbiological culture Collection center (China Committee for culture Collection of microorganisms) at the year 6 and the day 1, and the strain is named as China center for microorganisms (China) institute, national institute of sciences, no. 3, north Chen West road No.1, the area of towards the sun, beijing, and the classification name is: escherichia coli.
The preservation number of the lysine production strain MHZ-0913-3 is CGMCC No.11942, and the strain is preserved in the China general microbiological culture Collection center of China Committee for culture Collection of microorganisms for 12 months and 25 days, and is named as the national institute of microbiology, national institute of sciences, 1 st Caragana, 3 rd Chen, beijing, chaoyang, and classification: corynebacterium glutamicum Corynebacterium glutamicum, which strain is disclosed in patent CN 105734004B.
Example 1
This example is for illustrating an amino acid fermentation medium and a method for producing lysine by semi-continuous fermentation.
Inoculating lysine producing strain MHZ-0913-3 into the seed tank, and inoculating into the fermentation tank for fermentation when OD value reaches 0.8 after the seed tank is subjected to expansion culture.
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 2mg/L，FeSO 4 2mg/L, vitamin B1 100. Mu.g/L, biotin 200. Mu.g/L.
Fermentation conditions: the initial fermentation bottom sugar concentration is 2.5g/L, the ammonium sulfate nitrogen concentration is 2g/L, the fermentation temperature is 36-38 ℃, the fermentation pressure is controlled to be 0.08MPa, and the ventilation rate is 0.8m 3 /m 3 Continuously feeding fermentable sugar into a fermentation tank at a concentration of 58% by weight, mixing pure ammonium sulfate with extracted and recovered ammonium sulfate to prepare a solution according to a mass ratio of 9:1, wherein fed-batch amino acid nitrogen is threonine and methionine mixed solution, the concentration of residual sugar in the fermentation process is 3g/L, the concentration of fermented ammonia nitrogen is 1.5g/L, the fed-batch amino acid nitrogen accounts for 22% of the volume of the sugar solution, regulating and controlling the pH value by ammonia gas, and fermenting for 35h, wherein the pH value of fermentation is 6.6-6.8.
And discharging when the volume of the culture solution in the fermentation tank reaches 70% of the volume of the fermentation tank, wherein the discharging volume is 7% of the volume of the culture medium. The material is put into a secondary fermentation tank for continuous fermentation, the fermentation temperature is 36-38 ℃, the fermentation pressure is 0.06MPa, and the ventilation is 0.8m 3 /m 3 Continuously feeding fermentable sugar into a fermentation tank in a culture medium/min, wherein the concentration of the high-concentration sugar solution is 58%, mixing the fed-batch ammonium sulfate which is a pure ammonium sulfate product with the extracted and recovered ammonium sulfate according to the mass ratio of 9:1 to prepare a solution, feeding amino acid nitrogen which is threonine and methionine mixed solution, the concentration of residual sugar in the fermentation process is 5g/L, the concentration of fermented ammonia nitrogen is 1.2g/L, the fed-batch amino acid nitrogen accounts for 22% of the volume of the sugar solution, regulating and controlling the pH value by ammonia gas, and fermenting the pH value to be 6.7, and carrying out fermentation culture for 35h. After the fermentation is finished, the acid in the single batch tank and the continuous acid are measured, and the single batch conversion rate is calculated. The results are shown in Table 1.
Example 2
This example is for illustrating an amino acid fermentation medium and a method for producing lysine by semi-continuous fermentation.
Lysine was produced in the same manner as in example 1, except that the lysine-producing strain was MHZ-0914, the fermentation medium and the fermentation conditions were identical, and the culture broth in the fermenter was discharged when the volume of the culture broth was 75% of the volume of the fermenter, the discharge volume was 10% of the volume of the culture broth, the fermentation pressure was 0.05MPa, and the aeration rate was 1m 3 /m 3 Culture medium/min, other fed-batch nutrient conditions and examples1, and the fermentation time is 35h. After the fermentation is finished, the acid in the single batch tank and the continuous acid are measured, and the single batch conversion rate is calculated. The results are shown in Table 1.
Example 3
This example is for illustrating an amino acid fermentation medium and a method for producing lysine by semi-continuous fermentation.
Lysine was produced as in example 2, fermentation medium: glucose 19g/L, H3PO4 1.2g/L, KCl 0.4g/L, mgSO4 0.6g/L, molasses 9g/L, corn steep liquor 60g/L, soybean meal hydrolysate 30g/L, nicotinic acid 100mg/L, mnSO4 1.8mg/L, feSO4 1.8mg/L, vitamin B1 95 μg/L, biotin 190 μg/L.
The fermentation conditions, continuous release conditions and fed-batch nutrition conditions were the same as in example 2, and the fermentation time was 35h. After the fermentation is finished, the acid in the single batch tank and the continuous acid are measured, and the single batch conversion rate is calculated. The results are shown in Table 1.
Example 4
This example is for illustrating an amino acid fermentation medium and a method for producing lysine by semi-continuous fermentation.
Lysine was produced as in example 2, 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, 100mg/L nicotinic acid and MnSO 4 2mg/L，FeSO 4 2mg/L, vitamin B1 100. Mu.g/L, biotin 200. Mu.g/L.
The fermentation conditions, continuous release conditions and fed-batch nutrition conditions were the same as in example 2, and the fermentation time was 35h. After the fermentation is finished, the acid in the single batch tank and the continuous acid are measured, and the single batch conversion rate is calculated. The results are shown in Table 1.
Example 5
This example is for illustrating an amino acid fermentation medium and a method for producing lysine by semi-continuous fermentation.
Lysine was produced as in example 2, fermentation medium: glucose 20g/L, H 3 PO 4 1.5g/L，KCl 0.5g/L，MgSO 4 0.7g/L, molasses 10g/L, corn steep liquor 60g/L, beans30g/L of dreg hydrolysate, 120mg/L of nicotinic acid and MnSO 4 2mg/L，FeSO 4 2mg/L, vitamin B1 100. Mu.g/L, biotin 200. Mu.g/L.
The fermentation conditions, continuous release conditions and fed-batch nutrition conditions were the same as in example 2, and the fermentation time was 35h. After the fermentation is finished, the acid in the single batch tank and the continuous acid are measured, and the single batch conversion rate is calculated. The results are shown in Table 1.
Comparative example 1
Lysine was produced as in example 1, except that the medium contained no niacin and the material discharged during fermentation was directly placed in a continuous tank without further fermentation. After fermentation for 35 hours in the culture manner of example 1, acid in the single batch tank and acid in the continuous tank were measured, and the single batch conversion was calculated. The results are shown in Table 1.
TABLE 1
As can be seen from Table 1, the fermentation conversion rate is similar to that of the initial fermentation method, but the continuous acid content is greatly different, so that the continuous acid content and the fermentation conversion rate can be effectively improved by the method.
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 (9)
1. An amino acid fermentation medium, 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, 100-120mg/L nicotinic acid, mnSO 4 1.8-2.2mg/L，FeSO 4 1.8-2.2mg/L, vitamin B1 95-105 mug/L, biotin 190-210 mug/L.
2. A method for producing lysine by semi-continuous fermentation is characterized in that a lysine production strain is utilized, after the strain is subjected to seed tank expansion culture, the strain is inoculated into an amino acid fermentation culture medium in claim 1 for primary fermentation, when the volume of the culture medium in the primary fermentation tank reaches a certain proportion of the volume of the fermentation tank, part of fermentation liquor is periodically discharged, the amino acid fermentation culture medium is supplemented in the primary fermentation tank, the discharged fermentation liquor is fed into a secondary fermentation tank, secondary fermentation is continuously carried out by continuously feeding nutrients, the fermentation is finished, the secondary fermentation tank and the primary fermentation tank are discharged together, and the fermentation liquor enters a treatment flow of the fermentation liquor to obtain lysine.
3. The method for producing lysine by semi-continuous fermentation according to claim 2, wherein the lysine producing strain is MHZ-0914 and/or MHZ-0913-3, the preservation number of MHZ-0914 is CGMCC No.22648, and the preservation number of MHZ-0913-3 is CGMCC No.11942.
4. A method for producing lysine according to claim 3, wherein the lysine producing strain is MHZ-0914.
5. The method for producing lysine according to claim 2, wherein the fed-batch nutrient includes sugar, ammonium sulfate, threonine and methionine.
6. The method for producing lysine by semi-continuous fermentation according to claim 2, wherein the culture solution in the fermentation tank is discharged from the fermentation tank to 65% -75% of the volume of the fermentation tank, and the discharge volume is 5% -10% of the volume of the fermentation solution.
7. The method for producing lysine according to claim 2, wherein the secondary fermenter and the primary fermenter are provided with 1 or more.
8. The method for producing lysine by semi-continuous fermentation according to claim 2, wherein the secondary fermenter and the primary fermenter are each provided with a fed-batch system.
9. The method for producing lysine according to claim 2, wherein the secondary fermenter is fed without a medium, and fermentation is continued by feeding nutrients after feeding.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210726965.3A CN116287046A (en) | 2022-06-24 | 2022-06-24 | Amino acid fermentation medium and method for producing lysine by semi-continuous fermentation |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210726965.3A CN116287046A (en) | 2022-06-24 | 2022-06-24 | Amino acid fermentation medium and method for producing lysine by semi-continuous fermentation |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN116287046A true CN116287046A (en) | 2023-06-23 |
Family
ID=86813671
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202210726965.3A Pending CN116287046A (en) | 2022-06-24 | 2022-06-24 | Amino acid fermentation medium and method for producing lysine by semi-continuous fermentation |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN116287046A (en) |
-
2022
- 2022-06-24 CN CN202210726965.3A patent/CN116287046A/en active Pending
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN108220175B (en) | High-density culture method and pH regulation and control method for saccharomyces cerevisiae | |
| CN116333948B (en) | Clostridium aerophilum enrichment medium and preparation method thereof | |
| CN112501221A (en) | Method for improving conversion rate of threonine and saccharic acid | |
| CN115449499B (en) | Denitrification hyphomycete and method for preparing pyrroloquinoline quinone through fermentation of same | |
| CN116426443A (en) | Bacterial strain with quorum sensing behavior and application thereof in promotion of anaerobic ammonia oxidation denitrification | |
| FI71766C (en) | FRAMSTAELLNING AV ETHANOL GENOM HOEGEFFEKTIV BAKTERIEJAESNING. | |
| CN113502308B (en) | Method for producing vitamin B12 by aerobic fermentation based on redox potential regulation | |
| CN116287046A (en) | Amino acid fermentation medium and method for producing lysine by semi-continuous fermentation | |
| CN113528599B (en) | Production method of efficient chelating enzyme peptide | |
| CN114686531B (en) | Method for preparing 1,3-propanediol by biotransformation | |
| JP3074781B2 (en) | Production method of L-lysine by fermentation method | |
| CN112522338A (en) | Method for improving fermentation yield of L-tryptophan | |
| CN112852896A (en) | Fermentation production method of L-arginine | |
| CN114875090B (en) | Method for producing lysine and application thereof | |
| CN110885774A (en) | Method for optimizing glutamic acid fermentation | |
| CN110923275A (en) | Fermentation and extraction process of glutamic acid | |
| CN112662609B (en) | Fermentation medium for increasing yield of beta-alanine and application method | |
| CN117757863A (en) | Method for improving fermentation conversion rate of amino acid | |
| WO2007067005A1 (en) | Fermentation process for preparing l-lysine | |
| CN117431277A (en) | Method for producing L-amino acid by fermentation | |
| CN116218926A (en) | Method for producing acidic amino acid and application thereof | |
| CN117625706A (en) | Method for improving fermentation yield of L-proline | |
| CN113817627A (en) | Corynebacterium acetoacidophilum and method for performing glutamine fermentation under control of ammoniacal nitrogen concentration | |
| CN118308439A (en) | Novel efficient decoloring method for amino acid fermentation broth | |
| CN116286584A (en) | High osmotic pressure resistant strain screening method and application thereof in glutamic acid 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 |