CN111763636A - Method for producing coenzyme Q10 by industrial fermentation - Google Patents
Method for producing coenzyme Q10 by industrial fermentation Download PDFInfo
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- ACTIUHUUMQJHFO-UHFFFAOYSA-N Coenzym Q10 Natural products COC1=C(OC)C(=O)C(CC=C(C)CCC=C(C)CCC=C(C)CCC=C(C)CCC=C(C)CCC=C(C)CCC=C(C)CCC=C(C)CCC=C(C)CCC=C(C)C)=C(C)C1=O ACTIUHUUMQJHFO-UHFFFAOYSA-N 0.000 title claims abstract description 33
- 235000017471 coenzyme Q10 Nutrition 0.000 title claims abstract description 33
- ACTIUHUUMQJHFO-UPTCCGCDSA-N coenzyme Q10 Chemical compound COC1=C(OC)C(=O)C(C\C=C(/C)CC\C=C(/C)CC\C=C(/C)CC\C=C(/C)CC\C=C(/C)CC\C=C(/C)CC\C=C(/C)CC\C=C(/C)CC\C=C(/C)CCC=C(C)C)=C(C)C1=O ACTIUHUUMQJHFO-UPTCCGCDSA-N 0.000 title claims abstract description 32
- 229940110767 coenzyme Q10 Drugs 0.000 title claims abstract description 30
- 238000004519 manufacturing process Methods 0.000 title claims abstract description 21
- 238000009655 industrial fermentation Methods 0.000 title claims abstract description 7
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- 241000191043 Rhodobacter sphaeroides Species 0.000 claims abstract description 7
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- 239000000843 powder Substances 0.000 claims description 30
- 239000007788 liquid Substances 0.000 claims description 28
- 238000012258 culturing Methods 0.000 claims description 19
- 229910000402 monopotassium phosphate Inorganic materials 0.000 claims description 19
- 235000019796 monopotassium phosphate Nutrition 0.000 claims description 19
- 240000008042 Zea mays Species 0.000 claims description 18
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 claims description 18
- 235000002017 Zea mays subsp mays Nutrition 0.000 claims description 18
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 claims description 18
- 229910052921 ammonium sulfate Inorganic materials 0.000 claims description 18
- 235000011130 ammonium sulphate Nutrition 0.000 claims description 18
- 235000005822 corn Nutrition 0.000 claims description 18
- PXEDJBXQKAGXNJ-QTNFYWBSSA-L disodium L-glutamate Chemical compound [Na+].[Na+].[O-]C(=O)[C@@H](N)CCC([O-])=O PXEDJBXQKAGXNJ-QTNFYWBSSA-L 0.000 claims description 18
- SURQXAFEQWPFPV-UHFFFAOYSA-L iron(2+) sulfate heptahydrate Chemical compound O.O.O.O.O.O.O.[Fe+2].[O-]S([O-])(=O)=O SURQXAFEQWPFPV-UHFFFAOYSA-L 0.000 claims description 18
- 235000013923 monosodium glutamate Nutrition 0.000 claims description 18
- 229940073490 sodium glutamate Drugs 0.000 claims description 18
- 239000000463 material Substances 0.000 claims description 16
- WRUGWIBCXHJTDG-UHFFFAOYSA-L magnesium sulfate heptahydrate Chemical compound O.O.O.O.O.O.O.[Mg+2].[O-]S([O-])(=O)=O WRUGWIBCXHJTDG-UHFFFAOYSA-L 0.000 claims description 15
- 229940061634 magnesium sulfate heptahydrate Drugs 0.000 claims description 15
- 229940076230 magnesium sulfate monohydrate Drugs 0.000 claims description 15
- LFCFXZHKDRJMNS-UHFFFAOYSA-L magnesium;sulfate;hydrate Chemical compound O.[Mg+2].[O-]S([O-])(=O)=O LFCFXZHKDRJMNS-UHFFFAOYSA-L 0.000 claims description 15
- 238000009423 ventilation Methods 0.000 claims description 14
- 238000011081 inoculation Methods 0.000 claims description 13
- VTYYLEPIZMXCLO-UHFFFAOYSA-L Calcium carbonate Chemical compound [Ca+2].[O-]C([O-])=O VTYYLEPIZMXCLO-UHFFFAOYSA-L 0.000 claims description 12
- 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 12
- 239000002518 antifoaming agent Substances 0.000 claims description 12
- 239000008103 glucose Substances 0.000 claims description 12
- 239000013589 supplement Substances 0.000 claims description 12
- GNSKLFRGEWLPPA-UHFFFAOYSA-M potassium dihydrogen phosphate Chemical compound [K+].OP(O)([O-])=O GNSKLFRGEWLPPA-UHFFFAOYSA-M 0.000 claims description 11
- LWIHDJKSTIGBAC-UHFFFAOYSA-K potassium phosphate Substances [K+].[K+].[K+].[O-]P([O-])([O-])=O LWIHDJKSTIGBAC-UHFFFAOYSA-K 0.000 claims description 11
- OAICVXFJPJFONN-UHFFFAOYSA-N Phosphorus Chemical compound [P] OAICVXFJPJFONN-UHFFFAOYSA-N 0.000 claims description 9
- 229940041514 candida albicans extract Drugs 0.000 claims description 9
- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 claims description 9
- 238000010899 nucleation Methods 0.000 claims description 9
- 229910052698 phosphorus Inorganic materials 0.000 claims description 9
- 239000011574 phosphorus Substances 0.000 claims description 9
- 239000012138 yeast extract Substances 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 8
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 claims description 6
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 claims description 6
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 6
- 229910000019 calcium carbonate Inorganic materials 0.000 claims description 6
- 239000001110 calcium chloride Substances 0.000 claims description 6
- 229910001628 calcium chloride Inorganic materials 0.000 claims description 6
- GFHNAMRJFCEERV-UHFFFAOYSA-L cobalt chloride hexahydrate Chemical compound O.O.O.O.O.O.[Cl-].[Cl-].[Co+2] GFHNAMRJFCEERV-UHFFFAOYSA-L 0.000 claims description 6
- 229920001817 Agar Polymers 0.000 claims description 3
- 240000004808 Saccharomyces cerevisiae Species 0.000 claims description 3
- 239000008272 agar Substances 0.000 claims description 3
- 239000011790 ferrous sulphate Substances 0.000 claims description 3
- 235000003891 ferrous sulphate Nutrition 0.000 claims description 3
- 229910000359 iron(II) sulfate Inorganic materials 0.000 claims description 3
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- 229910052943 magnesium sulfate Inorganic materials 0.000 claims description 3
- 235000019341 magnesium sulphate Nutrition 0.000 claims description 3
- 238000009629 microbiological culture Methods 0.000 claims description 3
- 239000011780 sodium chloride Substances 0.000 claims description 3
- 238000012262 fermentative production Methods 0.000 claims 11
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- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 15
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- QGZKDVFQNNGYKY-UHFFFAOYSA-N Ammonia Chemical compound N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 description 4
- 239000002994 raw material Substances 0.000 description 4
- 239000008223 sterile water Substances 0.000 description 4
- 230000001954 sterilising effect Effects 0.000 description 4
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- 239000000047 product Substances 0.000 description 3
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- AFPLNGZPBSKHHQ-UHFFFAOYSA-N Betulaprenol 9 Natural products CC(C)=CCCC(C)=CCCC(C)=CCCC(C)=CCCC(C)=CCCC(C)=CCCC(C)=CCCC(C)=CCCC(C)=CCO AFPLNGZPBSKHHQ-UHFFFAOYSA-N 0.000 description 2
- 241001465754 Metazoa Species 0.000 description 2
- 241001052560 Thallis Species 0.000 description 2
- 229910021529 ammonia Inorganic materials 0.000 description 2
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- 230000001276 controlling effect Effects 0.000 description 2
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- AFPLNGZPBSKHHQ-MEGGAXOGSA-N solanesol Chemical compound CC(C)=CCC\C(C)=C\CC\C(C)=C\CC\C(C)=C\CC\C(C)=C\CC\C(C)=C\CC\C(C)=C\CC\C(C)=C\CC\C(C)=C\CO AFPLNGZPBSKHHQ-MEGGAXOGSA-N 0.000 description 2
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- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 1
- 208000024172 Cardiovascular disease Diseases 0.000 description 1
- 229920000742 Cotton Polymers 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 1
- 244000068988 Glycine max Species 0.000 description 1
- 235000010469 Glycine max Nutrition 0.000 description 1
- 244000061176 Nicotiana tabacum Species 0.000 description 1
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- 238000010521 absorption reaction Methods 0.000 description 1
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- 239000003963 antioxidant agent Substances 0.000 description 1
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- 235000006708 antioxidants Nutrition 0.000 description 1
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 1
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- 229940099596 manganese sulfate Drugs 0.000 description 1
- 239000011702 manganese sulphate Substances 0.000 description 1
- 235000007079 manganese sulphate Nutrition 0.000 description 1
- SQQMAOCOWKFBNP-UHFFFAOYSA-L manganese(II) sulfate Chemical compound [Mn+2].[O-]S([O-])(=O)=O SQQMAOCOWKFBNP-UHFFFAOYSA-L 0.000 description 1
- 230000002503 metabolic effect Effects 0.000 description 1
- 239000002207 metabolite Substances 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 229930014626 natural product Natural products 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- 235000016709 nutrition Nutrition 0.000 description 1
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- 230000003647 oxidation Effects 0.000 description 1
- 238000007254 oxidation reaction Methods 0.000 description 1
- 229910052760 oxygen Inorganic materials 0.000 description 1
- 239000001301 oxygen Substances 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 150000003254 radicals Chemical class 0.000 description 1
- NPCOQXAVBJJZBQ-UHFFFAOYSA-N reduced coenzyme Q9 Natural products COC1=C(O)C(C)=C(CC=C(C)CCC=C(C)CCC=C(C)CCC=C(C)CCC=C(C)CCC=C(C)CCC=C(C)CCC=C(C)CCC=C(C)C)C(O)=C1OC NPCOQXAVBJJZBQ-UHFFFAOYSA-N 0.000 description 1
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- 230000001502 supplementing effect Effects 0.000 description 1
- 229940035936 ubiquinone Drugs 0.000 description 1
- 210000001835 viscera Anatomy 0.000 description 1
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- 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
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- 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
- C12P7/00—Preparation of oxygen-containing organic compounds
- C12P7/66—Preparation of oxygen-containing organic compounds containing the quinoid structure
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Abstract
The invention relates to a preparation method of coenzyme Q10, in particular to a method for producing coenzyme Q10 by industrial fermentation. The method adopts rhodobacter sphaeroides as a strain, and comprises the following process steps of plate culture of the strain, slant culture of the strain, 20L seed culture, first-stage seed tank culture, second-stage seed culture, third-stage fermentation culture and seed pouring and re-culture fermentation. The invention solves the problems that the culture medium in the fermentation liquor is not completely consumed, the impurity content is high, the effective unit is low, the extraction yield is low, the quality of industrial salt is not easy to control, the low unit hyphae are more, the low yield and the pollution discharge are caused, and the COD discharge exceeds the standard due to more unused organic matters in the emission, thereby increasing the environmental protection cost. The method has the advantages of simple process flow, low risk of bacterial contamination, fermentation cost saving, fermentation yield improvement, process reserve increase and production discharge increase.
Description
Technical Field
The invention relates to a preparation method of coenzyme Q10, in particular to a method for producing coenzyme Q10 by industrial fermentation.
Background
Coenzyme Q10 (CoenzymeQ 10, CoQ 10), also known as ubiquinone, is a retinoid and is present in very low levels in cells of animals, plants, microorganisms, and the like. Coenzyme Q10 is a natural antioxidant and a cell metabolism activator synthesized by cells, can protect proteins, DNA molecules and the like from being damaged by free radical induced oxidation, and is widely applied to industries such as medicines, cosmetics, foods and the like. At present, coenzyme Q10 is a good clinical biochemical drug in medicine, and is commonly used for comprehensive treatment of liver diseases, cardiovascular diseases and cancers and improvement of immunity of organisms. The product is used as a novel food nutrition additive in the field of food, and has the functions of enhancing physical ability and improving immunity. Meanwhile, the coenzyme Q10 has irreplaceable effects on delaying senility and improving the immunity of the organism, and is widely applied to the field of female cosmetics.
The production method of coenzyme Q10 is generally classified into direct extraction, chemical synthesis, microbial fermentation, and the like. The direct extraction method is mainly used for separating and extracting soybean, tobacco leaf or animal viscera. The extraction method has simple preparation process, but has high extraction cost, is limited by raw materials, seasons and the like, and is not suitable for large-scale production in the modern chemical industry. The chemical synthesis methods are mainly classified into a full chemical synthesis method in which solanesol is not used as a raw material and a semi-chemical synthesis method in which solanesol is used as a raw material. The chemical synthesis method has the defects of complex reaction, multiple steps, low conversion efficiency and often existence of a plurality of byproducts, and the industrial development of the chemical synthesis method is influenced by the factors. The microbial fermentation method is the main method for producing coenzyme Q10 at present. The method has the advantages of cheap and rich raw materials, relatively simple product separation process, natural product, no compound chirality problem, good biological activity, easy absorption by human body, and realization of large-scale industrial production by fermentation tank, so the method becomes the coenzyme Q10 production method with the most development potential.
The normal fermentation period of the coenzyme Q10 is long, and nutrient substances such as carbon sources, nitrogen sources and the like need to be supplemented in order to meet the growth requirements of thalli in the fermentation process. However, the volume of the feed liquid in the fermentation tank gradually increases along with the increase of the feed supplement, so that the feed liquid overflows. In order to improve the equipment utilization rate of the fermentation tank, three times of intermediate discharge are required periodically. The volume of the fermentation broth per unit of batch is reduced to about one fourth of the total production per batch. The thalli with the discharging liquid does not reach the aging period, and the culture solution also contains partial nutrient substances which are not completely metabolized, for example, newly supplemented liquid can be discharged along with the discharging, so that the waste of raw and auxiliary materials is caused. From the fermentation period, the first two zones were in the logarithmic growth phase of the cells, and the third zone was in the cumulative peak phase of Q10. Therefore, the original process loses partial titer after 3 tape discharges. Meanwhile, the fermentation liquor with the fermentation liquor discharged for 3 times in the original process has a short period, the culture medium in the fermentation liquor is incompletely consumed, the impurity content is high, the effective unit is low, the extraction yield is low, the quality of industrial salt is not easy to control, low unit hyphae mostly cause low-yield pollution discharge, and unused organic matters in the emission mostly cause excessive COD discharge, so that the environmental protection cost is increased. Therefore, the improvement of the utilization rate of the low-unit fermentation liquor has great significance for increasing the yield, improving the technical index and reducing the cost.
Disclosure of Invention
In order to solve the problems in the prior art, the invention provides a method for producing coenzyme Q10 by industrial fermentation. Mainly collects the fermentation liquor with the sterile feed liquid in the tank to the receiving tank for secondary fermentation. The liquid with the material needs to provide part of the culture medium for the growth and the propagation of new hyphae and the synthesis of metabolites. The feeding opportunity and the feeding amount are timely regulated and controlled by detecting the indexes of the fermentation process, so that the hyphae keep a good production environment, and the aim of prolonging the growth period and improving the yield of the fermentation liquor is fulfilled. Because the fermentation liquor with the fermentation tubes is in the growth vigorous stage, the starting titer is high, and higher yield can be obtained in a shorter time. Meanwhile, as the fermentation liquor with the fermentation liquid is continuously operated for a certain period in the culture tank, nutrient substances in the culture medium are further consumed and utilized, and the filtration yield can be obviously improved due to the stable quality of hyphae, so that a certain effect is achieved on the stability of the product quality.
In order to realize the purpose, the invention provides a method for producing coenzyme Q10 by industrial fermentation, which adopts the following technical scheme:
the invention adopts Rhodobacter sphaeroides (Rhodobacter sphaeroides) which is preserved in China general microbiological culture Collection center in 24.10.2018, and the registration number of the preservation center is CGMCC No. 16625.
The method provided by the invention comprises the following steps:
s1: plate culture of strains: inoculating the strain on a flat plate, and culturing in an incubator with the temperature of 32.0 +/-0.5 ℃ and the humidity of 35-45% for 5-7 days;
s2: slant culture of strain; selecting mature strain from S1, culturing in incubator with temperature of 32.0 + -0.5 deg.C and humidity of 35% -45%, and culturing for 5-7 days;
s3: 20L of seeds are cultured; selecting strain matured by slant culture from S2, placing into sterilized 20L seeding tank, culturing at 60-80 rpm after 0-12 hr, 160-200 rpm after 13 hr, and air flow of 0.6 + -0.2 m3Culturing for 22-30 h at the temperature of 32.0 +/-1 ℃;
s4: first-level seeding tank culture: after the seed liquid in the S3 is cultured and matured, inoculating the seeds into a first-level seed tank by a pressure difference method according to the inoculation amount of 0.1-0.5 percent, wherein the first-level seed tank is cultured under the conditions that the tank pressure is 0.03-0.05 MPa and the ventilation volume is 100 +/-20 m3The temperature of the tank is 32-34 ℃, the rotating speed is 180-200 rpm, and the culture period is 24-32 hours;
s5: secondary seed culture: inoculating the mature primary seed liquid cultured in S4 into a secondary seed tank by a pressure difference method according to the inoculation amount of 8-20%, wherein the culture conditions of the secondary seed tank are that the tank pressure is 0.03-0.05 MPa, and the ventilation volume is 400 +/-200 m3The temperature of the tank is 32-34 ℃, the rotating speed is 120-160 rpm, and the culture period is 18-26 h;
s6: and (3) three-stage fermentation culture: inoculating the secondary seed liquid cultured and matured in S5 into a third-stage fermentation tank by a pressure difference method according to the inoculation amount of 20-30%, wherein the culture condition of the third-stage fermentation tank is that the tank pressure is 0.03-0.05 MPa, and the ventilation volume is controlled to be 800 +/-200 m after 40 h before fermentation3Per hour, after 40 hours to the end of ventilation control 1000 +/-200 m3The rotation speed is controlled to be 80-100 rpm, the tank temperature is 32-34 ℃, and 3 times of material supplement is needed in the fermentation process;
s7: pouring seeds and then culturingCulturing and fermenting: selecting 1 or more batches of fermentation liquor which is obtained in S6 and cultured for 50-65 h and meets the pure culture standard, transferring the fermentation liquor into a sterilized receiving tank for continuous culture, wherein the tank pressure of the receiving tank is 0.03-0.05 MPa, and the ventilation volume before fermentation is controlled to be 800 +/-200 m for 40 h3Per hour, after 40 hours to the end of ventilation control 1000 +/-200 m3And/h, controlling the rotating speed to be 80-100 rpm, controlling the tank temperature to be 32-34 ℃, and needing to perform 3 times of material supplement in the fermentation process.
Further, the culture medium formula for the S1 plate culture and the step S2 slant culture is as follows: 1.0-10.0 g/L of yeast powder, 0.1-2.5 g/L of monopotassium phosphate, 1.0-5.0 g/L of magnesium sulfate, 0.1-1.0 g/L of ferrous sulfate, 0-5.0 g/L of sodium chloride, 1.5-3.5 g/L of ammonium sulfate, 0.5-3.0g/L of sodium glutamate, 0.5-3.0g/L of corn steep liquor dry powder, 5-20g/L of glucose, 15-20 g/L of agar and 6.5-7.3 of pH.
Further, the culture tank medium formula for 20L of seed culture in S3 is as follows: 30-60 g/L of glucose, 10-30 g/L of ammonium sulfate, 5-10 g/L of corn steep liquor dry powder, 1-10 g/L of sodium glutamate, 0.1-0.5 g/L of magnesium sulfate monohydrate, 5-20g/L of yeast extract powder, 15-30 g/L of industrial salt, 5-10 g/L of dipotassium hydrogen phosphate, 5-10 g/L of potassium dihydrogen phosphate, 10-30 g/L of magnesium sulfate heptahydrate, 1-10 g/L of ferrous sulfate heptahydrate, and the pH = 6.5-7.3.
Further, the formula of the first-level seed tank culture medium in the S4 is as follows: 0.1-0.4% of glucose, 0.2-0.5% of ammonium sulfate, 0.03-0.06% of dipotassium hydrogen phosphate, 0.03-0.06% of monopotassium phosphate, 0.1-0.3% of magnesium sulfate heptahydrate, 0.01-0.03% of ferrous sulfate heptahydrate, 0.1-0.4% of industrial salt, 0.5-1.0% of calcium carbonate, 0.01-0.05% of defoaming agent, 0.03-0.08% of corn steep liquor dry powder, 0.03-0.08% of sodium glutamate, 0.001-0.005% of magnesium sulfate monohydrate, 0.0001-0.0005% of cobalt chloride hexahydrate, 0.1-0.5% of yeast extract powder and pH = 6.5-7.3.
Further, the formula of the secondary seed tank culture medium in the S5 is as follows: 1-2% of glucose, 0.1-0.5% of ammonium sulfate, 0.05-1.0% of dipotassium hydrogen phosphate, 0.05-1.0% of monopotassium phosphate, 0.1-0.5% of magnesium sulfate heptahydrate, 0.01-0.04% of ferrous sulfate heptahydrate, 0.1-0.5% of industrial salt, 0.5-1.0% of calcium carbonate, 0.01-0.05% of defoaming agent, 0.05-0.15% of corn steep liquor dry powder, 0.05-0.1% of sodium glutamate, 0.005-0.015% of magnesium sulfate monohydrate, 0.0001-0.0005% of cobalt chloride hexahydrate, 0.1-0.5% of yeast extract powder and pH = 6.5-7.3.
Further, the culture medium formula of the third-stage fermentation tank in the S6 third-stage fermentation culture is as follows: 0.2 to 0.6 percent of ammonium sulfate, 0.01 to 0.06 percent of monopotassium phosphate, 1.0 to 1.5 percent of magnesium sulfate heptahydrate, 0.1 to 0.5 percent of ferrous sulfate heptahydrate, 0.1 to 0.5 percent of industrial salt, 0.01 to 0.05 percent of defoaming agent, 0.005 to 0.01 percent of calcium chloride, 0.3 to 0.8 percent of corn steep liquor dry powder, 0.1 to 0.5 percent of sodium glutamate, 0.005 to 0.01 percent of magnesium sulfate monohydrate, and the pH =6.5 to 7.3.
Further, the determination criterion of the fermentation end point in S4 is as follows: the OD of the fermentation liquor is more than or equal to 5.0, and the concentration of reducing sugar is reduced to 0-0.5 g/L.
Further, the determination criterion of the fermentation end point in S5 is as follows: the OD of the fermentation liquor is more than or equal to 10.0, and the concentration of reducing sugar is reduced to 0-0.5 g/L.
Further, samples were taken every 4 hours after inoculation in S6 and S7 to determine sugar, phosphorus, and microscopically observe bacterial forms and monitor bacterial contamination.
Further, the sugar concentration is controlled to be 0.8-1.2% 40 h before the three-stage fermentation of S6 and the reseeding and re-culturing fermentation of S7, the sugar concentration is controlled to be 0.5-0.8% 40-80 h in the middle stage of fermentation, the sugar concentration is controlled to be about 0.2% after 80 h and to be finished, the phosphorus supplement speed is controlled to be 35-40L/h 4-40 h before fermentation, 10-15L/h 40-65 h and 25-30L/h 65-80 h.
Furthermore, 3 times of feeding are required in the processes of tertiary fermentation of S6 and reseeding and re-culturing fermentation of S7, the first feeding time is 20-30 hours, the second feeding time is 40-50 hours, and the third feeding time is 90-105 hours of the culture period before reseeding.
Further, the feed formula of the S6 tertiary fermentation and the S7 reverse seed re-culture fermentation process is as follows: 0.5 to 1.0 percent of ammonium sulfate, 0.1 to 0.5 percent of monopotassium phosphate, 1.5 to 3.5 percent of magnesium sulfate heptahydrate, 0.1 to 0.5 percent of ferrous sulfate heptahydrate, 0.1 to 0.3 percent of defoaming agent, 0.01 to 0.03 percent of calcium chloride, 1.0 to 3.0 percent of corn steep liquor dry powder, 0.5 to 1.0 percent of sodium glutamate and 0.01 to 0.03 percent of magnesium sulfate monohydrate.
The beneficial effects obtained by the invention are as follows:
1) the process flow is simplified, and the risk of bacterial contamination is reduced: the prior art comprises about 60 hours and about 70 hours before the third feeding respectively. And 3 times of band amplification of the original process is simplified into 1 time of band amplification by adopting a band amplification re-culture process, and meanwhile, the process of seed culture is reduced. Not only simplifies the process flow, but also reduces the transfer times of the seed liquid and the fermentation liquid and reduces the contamination probability.
2) Save the fermentation cost, promote fermentation output: the three times of the prior art are directly put into the extraction section for extraction. The three times of belt discharge can cause insufficient utilization of the culture medium, and if newly supplemented feed liquid is discharged along with the belt discharge, the waste of raw and auxiliary materials is caused. From the fermentation period, the first two zones were in the logarithmic growth phase of the cells, and the third zone was in the cumulative peak phase of Q10. Therefore, the original process loses partial titer after 3 tape discharges. By adopting the area-releasing re-culture process, the material loss of the original area-releasing process can be saved, and the fermentation titer can be improved by prolonging the metabolic growth period of the strain through the culture area-releasing.
3) Is beneficial to environmental protection: because the fermentation liquor with the fermentation liquor discharged for 3 times in the original process has a short period, the culture medium in the fermentation liquor is incompletely consumed, the impurity content is high, the effective unit is low, the extraction yield is low, the quality of industrial salt is not easy to control, low unit hyphae mostly cause low-yield pollution discharge, and unused organic matters in the emission mostly cause excessive COD discharge, so that the environmental protection cost is increased. The above problems can be improved by using the strip re-culture process.
4) Increase of process reserves, increase of production scheduling: the development of the re-culture process with the placing is added with process selection for a workshop, so that a proper process can be conveniently selected according to the actual fermentation condition. Meanwhile, the implementation of the re-culture process with releasing is beneficial to the production scheduling of workshops, and the production batch per month can be increased to a certain extent.
Drawings
FIG. 1 is a flow chart of the fermentation process of the present invention;
FIG. 2 is a graph comparing the fermentation indexes of the "one-by-two" inverted seeding process and the original process in example 1;
FIG. 3 is a graph comparing the fermentation indexes of the "two-band three-band" inverted planting process and the original process in example 2.
Detailed Description
The invention will be further described with reference to specific examples, but the scope of the invention is not limited thereto.
The flow chart of the fermentation process of the invention is shown in figure 1.
The invention adopts Rhodobacter sphaeroides (Rhodobacter sphaeroides) which is preserved in China general microbiological culture Collection center in 24.10.2018, and the registration number of the preservation center is CGMCC No. 16625.
The media formulations used in examples 1-2 were as follows:
plate and slant medium (medium a): 10.0 g/L of yeast powder, 2.5 g/L of monopotassium phosphate, 2.0 g/L of magnesium sulfate, 0.02 g/L of ferrous sulfate, 0.03 g/L of manganese sulfate, 2.0 g/L of sodium chloride, 2.0 g/L of ammonium sulfate, 1.0g/L of sodium glutamate, 2.0 g/L of corn steep liquor dry powder, 10 g/L of glucose and 15-20 g/L of agar.
20L of seeding tank medium (Medium B): 50 g/L of glucose, 20g/L of ammonium sulfate, 5g/L of corn steep liquor dry powder, 5g/L of sodium glutamate, 0.3 g/L of magnesium sulfate monohydrate, 10 g/L of yeast extract powder, 20g/L of industrial salt, 5.0 g/L of dipotassium hydrogen phosphate, 5.0 g/L of potassium dihydrogen phosphate, 20g/L of magnesium sulfate heptahydrate and 1.0g/L of ferrous sulfate heptahydrate.
Primary seed tank medium (medium C): 0.4% of glucose, 0.25% of ammonium sulfate, 0.05% of dipotassium hydrogen phosphate, 0.05% of potassium dihydrogen phosphate, 0.2% of magnesium sulfate heptahydrate, 0.01% of ferrous sulfate heptahydrate, 0.2% of industrial salt, 0.8% of calcium carbonate, 0.03% of defoaming agent, 0.05% of corn steep liquor dry powder, 0.05% of sodium glutamate, 0.003% of magnesium sulfate monohydrate, 0.0001% of cobalt chloride hexahydrate and 0.1% of yeast extract powder.
Secondary seed tank medium (medium D): 1.5 percent of glucose, 0.3 percent of ammonium sulfate, 0.08 percent of dipotassium hydrogen phosphate, 0.08 percent of potassium dihydrogen phosphate, 0.35 percent of magnesium sulfate heptahydrate, 0.02 percent of ferrous sulfate heptahydrate, 0.2 percent of industrial salt, 0.8 percent of calcium carbonate, 0.02 percent of defoaming agent, 0.1 percent of corn steep liquor dry powder, 0.08 percent of sodium glutamate, 0.01 percent of magnesium sulfate monohydrate, 0.00015 percent of cobalt chloride hexahydrate and 0.2 percent of yeast extract powder.
Third stage fermenter Medium (Medium E): 0.415 percent of ammonium sulfate, 0.05 percent of potassium dihydrogen phosphate, 1.3 percent of magnesium sulfate heptahydrate, 0.14 percent of ferrous sulfate heptahydrate, 0.25 percent of industrial salt, 0.015 percent of defoaming agent, 0.008 percent of calcium chloride, 0.4 percent of corn steep liquor dry powder, 0.3 percent of sodium glutamate and 0.006 percent of magnesium sulfate monohydrate.
The formula of the feed supplement is as follows: 0.65% of ammonium sulfate, 0.2% of potassium dihydrogen phosphate, 2.3% of magnesium sulfate heptahydrate, 0.21% of ferrous sulfate heptahydrate, 0.1% of defoaming agent, 0.01% of calcium chloride, 2.0% of corn steep liquor dry powder, 0.6% of sodium glutamate and 0.01% of magnesium sulfate monohydrate.
The above culture medium was sterilized at 121 deg.C for 30 min.
Sugar supplementing and sterilizing conditions: 60% liquid sugar is sterilized at 115.5 + -0.5 deg.C for 25 min.
Liquid phosphorus sterilization conditions: 0.08 g/L potassium dihydrogen phosphate is sterilized at 121 +/-1 ℃ for 30 min.
Example 1 "two-in-one" seed inversion method
Plate culture of S1 strain: and (4) preparing a culture medium according to the formula of the culture medium A, sterilizing, and pouring the plate under the aseptic condition. And (3) sucking 1 mL of glycerol bacterial suspension by using a pipette gun, adding the glycerol bacterial suspension into a test tube filled with 9 mL of sterile water, oscillating and scattering the glycerol bacterial suspension on a vortex oscillator, uniformly mixing the glycerol bacterial suspension and the sterile water, and diluting the glycerol bacterial suspension by using the pipette gun. With dilution of 10-5The bacterial suspension of (1) was inoculated into plates, and 200-300. mu.L of bacterial suspension was placed in each plate and then spread evenly using a sterilized spreader. Putting the evenly coated flat plate into an incubator with the temperature of 32.0 +/-0.5 ℃ and the humidity of 35-45 percent for culturing for 5-7 days.
Slant culture of S2 strain: after preparing the culture medium according to the formula of the culture medium A and sterilizing the culture medium, the inclined plane is inclined under the aseptic condition. Selecting a plurality of single colonies which are full, convex and uniform in size from a mother plate which is mature in culture by using an inoculation shovel, transferring the single colonies into a test tube filled with 9 mL of sterile water, vibrating the single colonies on a vortex oscillator, scattering the single colonies, uniformly mixing the single colonies, and diluting the single colonies to 10 times by using a pipette-5. The bacterial suspension was pipetted with a 0.5mL pipette and spread evenly into the tube. Culturing in an incubator with temperature of 32.0 + -0.5 deg.C and humidity of 35% -45% for 5-7 days.
S3: 20L of seed culture: taking 1-3 inclined planes, disinfecting the outer wall of the test tube with alcohol cotton, selecting a plurality of single bacterial colonies which are full, convex and uniform in size with an inoculation shovel, transferring the single bacterial colonies into a sterile water test tube with 50-60 mL for sufficient scattering, and transferring the bacterial suspension into a sterilized 20L seed tankThe culture conditions are that the rotating speed is controlled to be 60-80 rpm after 0-12 h, the rotating speed is controlled to be 160-200 rpm after 13 h, and the air flow is 0.6 +/-0.2 m3And culturing for 22-30 h at the temperature of 32.0 +/-1 ℃.
S4: first-level seeding tank culture: after 20L of seed liquid is cultured to be mature, inoculating the mature seed liquid into a first-level seed tank by a pressure difference method according to the inoculation amount of 0.1-0.5%, wherein the first-level seed tank culture conditions are that the tank pressure is 0.03-0.05 MPa, the ventilation volume is 100 +/-20 m3/h, the tank temperature is 32-34 ℃, the rotation speed is 180-200 rpm, and the culture period is 24-32 h. And (3) judging the fermentation end point standard: the OD of the fermentation liquor is more than or equal to 5.0, and the concentration of reducing sugar is reduced to 0-0.5 g/L.
S5: secondary seed culture: inoculating the first-level seed liquid which is mature in culture and free of infectious microbes into a second-level seed tank by a pressure difference method according to the inoculation amount of 8-20%, wherein the culture conditions of the second-level seed tank are that the tank pressure is 0.03-0.05 MPa, and the ventilation volume is 400 +/-200 m3The temperature of the tank is 32-34 ℃, the rotating speed is 120-160 rpm, and the culture period is 18-26 h. And (3) judging the fermentation end point standard: the OD of the fermentation liquor is more than or equal to 10.0, and the concentration of reducing sugar is reduced to 0-0.5 g/L.
S6: the 'one-to-two' seed reversing and re-culturing process comprises the following steps: and (3) inoculating the secondary seed liquid which is mature in culture and free of the mixed bacteria into a third-stage fermentation tank by a pressure difference method according to the inoculation amount of 20-30%, selecting the fermentation tank which is subjected to fermentation culture for 45-48 h and meets the pure culture standard as a fermentation liquid delivery tank, and uniformly distributing the fermentation liquid into another sterilized receiving tank for continuous culture. As can be seen from the attached figure 2, the titer of the tank discharge of the 'one-by-two' seed pouring process is basically equal to that of the original process. Because the fermentation process of the 'one-by-two' seed pouring process is not carried out for 3 times of discharging, the volume of the discharging tank is 30 m less than that of the original process3Correspondingly, the total billion of the tank placing is slightly lower than that of the original process, which indirectly proves that the coenzyme Q10 content of the three-time material placing liquid of the original process is lower. Compared with the original process, the inverted process with two batches can reduce the culture of one batch of seed tanks at different levels and the preparation of one batch of basic materials of a fermentation tank under the condition that the unit consumption of sugar is basically consistent, so that the average phosphorus and ammonia supplementation amount of each batch is relatively reduced, the workload is reduced, and the cost is reduced. Meanwhile, the implementation of the re-culture process with releasing is beneficial to the production scheduling of workshops, the production batch per month can be increased to a certain extent, and the equipment utilization rate and the production rate are greatly improvedAnd (4) fermentation yield.
Specific process controls are shown in the following table.
'one-to-two' seed reversing process control
Example 2 implementation of the "two-zone three" seed inversion Process
The early stage fermentation process of 'flat seeds-inclined plane seeds-20L seeds-first-stage seeds-second-stage seeds' is basically controlled in a consistent way. And in the 'two-band three' seed-reversing re-culture process, a fermentation tank which is subjected to fermentation culture for 45-65 hours and meets the pure seed culture standard is selected as a fermentation liquid feeding tank, and the fermentation liquid in the fermentation tank is uniformly transferred into another sterilized receiving tank for continuous culture. As can be seen from the attached figure 3, the tank-placing titer of the 'two-belt three-tank' inverted seeding process is slightly higher than that of the original process, and mainly after two-belt tank-placing inverted seeding, the volume of fermentation liquor is reduced, so that the charging coefficient of a fermentation tank is effectively improved, the fermentation dissolved oxygen level is improved, and the tank-placing titer is promoted to be improved. Also, the fermentation process of the 'two-belt three-belt' seed pouring process is not carried out for 3 times of belt discharging, so the tank discharging volume is reduced by 27 m compared with the original process3Correspondingly, the total billion of the tank placing is slightly lower than that of the original process, which indirectly proves that the coenzyme Q10 content of the three-time material placing liquid of the original process is lower. Compared with the original process, the 'two-belt three-seed' inverted process can reduce the culture of a batch of seed tanks at different levels and the preparation of a batch of basic materials of a fermentation tank under the condition that the unit consumption of sugar is basically consistent, and meanwhile, the average phosphorus and ammonia supplementation amount of each batch is relatively reduced, so that the workload is reduced and the cost is reduced. Meanwhile, the implementation of the re-culture process with the placement is beneficial to the production scheduling of workshops, the production batch per month can be increased to a certain extent, and the equipment utilization rate and the fermentation yield are greatly improved.
Specific process controls are shown in the following table.
Claims (12)
1. A method for producing coenzyme Q10 by industrial fermentation is characterized in that,
the adopted strain is Rhodobacter sphaeroides (Rhodobacter sphaeroides), which is preserved in China general microbiological culture Collection center in 24/10.2018, and the registration number of the preservation center is CGMCC No. 16625;
the method comprises the following steps:
s1: plate culture of strains: inoculating the strain on a flat plate, and culturing in an incubator with the temperature of 32.0 +/-0.5 ℃ and the humidity of 35-45% for 5-7 days;
s2: slant culture of strain; selecting mature strain from S1, culturing in incubator with temperature of 32.0 + -0.5 deg.C and humidity of 35% -45%, and culturing for 5-7 days;
s3: 20L of seeds are cultured; selecting strain matured by slant culture from S2, placing into sterilized 20L seeding tank, culturing at 60-80 rpm after 0-12 hr, 160-200 rpm after 13 hr, and air flow of 0.6 + -0.2 m3Culturing for 22-30 h at the temperature of 32.0 +/-1 ℃;
s4: first-level seeding tank culture: after the seed liquid in the S3 is cultured and matured, inoculating the seeds into a first-level seed tank by a pressure difference method according to the inoculation amount of 0.1-0.5 percent, wherein the first-level seed tank is cultured under the conditions that the tank pressure is 0.03-0.05 MPa and the ventilation volume is 100 +/-20 m3The temperature of the tank is 32-34 ℃, the rotating speed is 180-200 rpm, and the culture period is 24-32 hours;
s5: secondary seed culture: inoculating the mature primary seed liquid cultured in S4 into a secondary seed tank by a pressure difference method according to the inoculation amount of 8-20%, wherein the culture conditions of the secondary seed tank are that the tank pressure is 0.03-0.05 MPa, and the ventilation volume is 400 +/-200 m3The temperature of the tank is 32-34 ℃, the rotating speed is 120-160 rpm, and the culture period is 18-26 h;
s6: and (3) three-stage fermentation culture: inoculating the secondary seed liquid cultured and matured in S5 into a third-stage fermentation tank by a pressure difference method according to the inoculation amount of 20-30%, wherein the culture condition of the third-stage fermentation tank is that the tank pressure is 0.03-0.05 MPa, and the ventilation volume is controlled to be 800 +/-200 m after 40 h before fermentation3Per hour, after 40 hours to the end of ventilation control 1000 +/-200 m3The rotation speed is controlled to be 80-100 rpm, the tank temperature is 32-34 ℃, and 3 times of material supplement is needed in the fermentation process;
s7: seed fallingAnd (3) re-culturing and fermenting: selecting 1 or more batches of fermentation liquor which is obtained in S6 and cultured for 50-65 h and meets the pure culture standard, transferring the fermentation liquor into a sterilized receiving tank for continuous culture, wherein the tank pressure of the receiving tank is 0.03-0.05 MPa, and the ventilation volume before fermentation is controlled to be 800 +/-200 m for 40 h3Per hour, after 40 hours to the end of ventilation control 1000 +/-200 m3And/h, controlling the rotating speed to be 80-100 rpm, controlling the tank temperature to be 32-34 ℃, and needing to perform 3 times of material supplement in the fermentation process.
2. The method for the industrial fermentative production of coenzyme Q10 according to claim 1, characterized in that: the culture medium formula used for the S1 plate culture and the S2 slant culture is as follows: 1.0-10.0 g/L of yeast powder, 0.1-2.5 g/L of monopotassium phosphate, 1.0-5.0 g/L of magnesium sulfate, 0.1-1.0 g/L of ferrous sulfate, 0-5.0 g/L of sodium chloride, 1.5-3.5 g/L of ammonium sulfate, 0.5-3.0g/L of sodium glutamate, 0.5-3.0g/L of corn steep liquor dry powder, 5-20g/L of glucose, 15-20 g/L of agar and 6.5-7.3 of pH.
3. The method for the industrial fermentative production of coenzyme Q10 according to claim 1, characterized in that: the culture medium formula of the culture tank for culturing 20L of seeds in S3 is as follows: 30-60 g/L of glucose, 10-30 g/L of ammonium sulfate, 5-10 g/L of corn steep liquor dry powder, 1-10 g/L of sodium glutamate, 0.1-0.5 g/L of magnesium sulfate monohydrate, 5-20g/L of yeast extract powder, 15-30 g/L of industrial salt, 5-10 g/L of dipotassium hydrogen phosphate, 5-10 g/L of potassium dihydrogen phosphate, 10-30 g/L of magnesium sulfate heptahydrate, 1-10 g/L of ferrous sulfate heptahydrate, and the pH = 6.5-7.3.
4. The method for the industrial fermentative production of coenzyme Q10 according to claim 1, characterized in that: the formula of the culture medium of the first-level seed tank in the S4 is as follows: 0.1-0.4% of glucose, 0.2-0.5% of ammonium sulfate, 0.03-0.06% of dipotassium hydrogen phosphate, 0.03-0.06% of monopotassium phosphate, 0.1-0.3% of magnesium sulfate heptahydrate, 0.01-0.03% of ferrous sulfate heptahydrate, 0.1-0.4% of industrial salt, 0.5-1.0% of calcium carbonate, 0.01-0.05% of defoaming agent, 0.03-0.08% of corn steep liquor dry powder, 0.03-0.08% of sodium glutamate, 0.001-0.005% of magnesium sulfate monohydrate, 0.0001-0.0005% of cobalt chloride hexahydrate, 0.1-0.5% of yeast extract powder and pH = 6.5-7.3.
5. The method for the industrial fermentative production of coenzyme Q10 according to claim 1, characterized in that: the formula of the secondary seed tank culture medium in the S5 is as follows: 1-2% of glucose, 0.1-0.5% of ammonium sulfate, 0.05-1.0% of dipotassium hydrogen phosphate, 0.05-1.0% of monopotassium phosphate, 0.1-0.5% of magnesium sulfate heptahydrate, 0.01-0.04% of ferrous sulfate heptahydrate, 0.1-0.5% of industrial salt, 0.5-1.0% of calcium carbonate, 0.01-0.05% of defoaming agent, 0.05-0.15% of corn steep liquor dry powder, 0.05-0.1% of sodium glutamate, 0.005-0.015% of magnesium sulfate monohydrate, 0.0001-0.0005% of cobalt chloride hexahydrate, 0.1-0.5% of yeast extract powder and pH = 6.5-7.3.
6. The method for the industrial fermentative production of coenzyme Q10 according to claim 1, characterized in that: the culture medium formula of the third-stage fermentation tank in the S6 third-stage fermentation culture is as follows: 0.2 to 0.6 percent of ammonium sulfate, 0.01 to 0.06 percent of monopotassium phosphate, 1.0 to 1.5 percent of magnesium sulfate heptahydrate, 0.1 to 0.5 percent of ferrous sulfate heptahydrate, 0.1 to 0.5 percent of industrial salt, 0.01 to 0.05 percent of defoaming agent, 0.005 to 0.01 percent of calcium chloride, 0.3 to 0.8 percent of corn steep liquor dry powder, 0.1 to 0.5 percent of sodium glutamate, 0.005 to 0.01 percent of magnesium sulfate monohydrate, and the pH =6.5 to 7.3.
7. The method for the industrial fermentative production of coenzyme Q10 according to claim 1, characterized in that: the judgment standard of the fermentation end point in the S4 is as follows: the OD of the fermentation liquor is more than or equal to 5.0, and the concentration of reducing sugar is reduced to 0-0.5 g/L.
8. The method for the industrial fermentative production of coenzyme Q10 according to claim 1, characterized in that: the judgment standard of the fermentation end point in the S5 is as follows: the OD of the fermentation liquor is more than or equal to 10.0, and the concentration of reducing sugar is reduced to 0-0.5 g/L.
9. The method for the industrial fermentative production of coenzyme Q10 according to claim 1, characterized in that: samples were taken every 4 h after inoculation in S6 and S7 to determine sugar, phosphorus, microscopically observe bacterial form and monitor bacterial contamination.
10. The method for the industrial fermentative production of coenzyme Q10 according to claim 9, characterized in that: the sugar concentration is controlled to be 0.8-1.2% 40 h before the three-stage fermentation of S6 and the reverse-seed re-culture fermentation of S7, the sugar concentration is controlled to be 0.5-0.8% 40-80 h in the middle stage of fermentation, the sugar concentration is controlled to be about 0.2% after 80 h and to be finished, the phosphorus supplement speed is controlled to be 35-40L/h 4-40 h before fermentation, the phosphorus supplement speed is controlled to be 10-15L/h 40-65 h, and the phosphorus supplement speed is controlled to be 25-30L/h 65-80 h.
11. The method for the industrial fermentative production of coenzyme Q10 according to claim 10, characterized in that: the three-stage fermentation of S6 and the reseeding and re-culturing fermentation process of S7 require 3 times of material supplement, the first time of material supplement lasts for 20-30 hours, the second time of material supplement lasts for 40-50 hours, and the third time of material supplement lasts for 90-105 hours before reseeding.
12. The method for the industrial fermentative production of coenzyme Q10 according to claim 11, characterized in that: the feed formula for the S6 tertiary fermentation and the S7 seed-pouring re-culture fermentation process is as follows: 0.5 to 1.0 percent of ammonium sulfate, 0.1 to 0.5 percent of monopotassium phosphate, 1.5 to 3.5 percent of magnesium sulfate heptahydrate, 0.1 to 0.5 percent of ferrous sulfate heptahydrate, 0.1 to 0.3 percent of defoaming agent, 0.01 to 0.03 percent of calcium chloride, 1.0 to 3.0 percent of corn steep liquor dry powder, 0.5 to 1.0 percent of sodium glutamate and 0.01 to 0.03 percent of magnesium sulfate monohydrate.
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| CN115125177A (en) * | 2022-08-02 | 2022-09-30 | 北大方正集团有限公司 | Fermentation method and method for producing coenzyme Q10 by fermentation |
| CN115125177B (en) * | 2022-08-02 | 2024-04-26 | 北大方正集团有限公司 | Fermentation method and method for producing coenzyme Q10 by fermentation |
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