CN105483171B - A kind of raising ubiquinone10The production method of industrial output - Google Patents
A kind of raising ubiquinone10The production method of industrial output Download PDFInfo
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- CN105483171B CN105483171B CN201511031083.1A CN201511031083A CN105483171B CN 105483171 B CN105483171 B CN 105483171B CN 201511031083 A CN201511031083 A CN 201511031083A CN 105483171 B CN105483171 B CN 105483171B
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- 238000004519 manufacturing process Methods 0.000 title claims abstract description 32
- 238000000855 fermentation Methods 0.000 claims abstract description 75
- 230000004151 fermentation Effects 0.000 claims abstract description 74
- 239000002609 medium Substances 0.000 claims abstract description 43
- 230000001580 bacterial effect Effects 0.000 claims abstract description 35
- 239000001963 growth medium Substances 0.000 claims abstract description 29
- 238000011218 seed culture Methods 0.000 claims abstract description 23
- 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 claims abstract description 20
- 238000012216 screening Methods 0.000 claims abstract description 20
- 238000011084 recovery Methods 0.000 claims abstract description 4
- XEEYBQQBJWHFJM-UHFFFAOYSA-N iron Substances [Fe] XEEYBQQBJWHFJM-UHFFFAOYSA-N 0.000 claims description 54
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 claims description 24
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 20
- 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 15
- 239000008103 glucose Substances 0.000 claims description 15
- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 claims description 12
- 229910000359 iron(II) sulfate Inorganic materials 0.000 claims description 12
- 229910052943 magnesium sulfate Inorganic materials 0.000 claims description 12
- 235000019341 magnesium sulphate Nutrition 0.000 claims description 12
- 238000007747 plating Methods 0.000 claims description 12
- 239000011790 ferrous sulphate Substances 0.000 claims description 11
- 235000003891 ferrous sulphate Nutrition 0.000 claims description 11
- BAUYGSIQEAFULO-UHFFFAOYSA-L iron(2+) sulfate (anhydrous) Chemical compound [Fe+2].[O-]S([O-])(=O)=O BAUYGSIQEAFULO-UHFFFAOYSA-L 0.000 claims description 11
- 150000001875 compounds Chemical group 0.000 claims description 10
- 239000004615 ingredient Substances 0.000 claims description 10
- 239000011780 sodium chloride Substances 0.000 claims description 10
- 229940041514 candida albicans extract Drugs 0.000 claims description 9
- 239000012138 yeast extract Substances 0.000 claims description 9
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 claims description 8
- 229910052921 ammonium sulfate Inorganic materials 0.000 claims description 8
- 235000011130 ammonium sulphate Nutrition 0.000 claims description 8
- FBAFATDZDUQKNH-UHFFFAOYSA-M iron chloride Chemical compound [Cl-].[Fe] FBAFATDZDUQKNH-UHFFFAOYSA-M 0.000 claims description 2
- KRKNYBCHXYNGOX-UHFFFAOYSA-K Citrate Chemical compound [O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O KRKNYBCHXYNGOX-UHFFFAOYSA-K 0.000 claims 1
- 238000000034 method Methods 0.000 abstract description 33
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 48
- 241000894006 Bacteria Species 0.000 description 25
- 229910052742 iron Inorganic materials 0.000 description 23
- 230000008569 process Effects 0.000 description 16
- 230000001954 sterilising effect Effects 0.000 description 13
- 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 description 12
- 235000017471 coenzyme Q10 Nutrition 0.000 description 12
- 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 12
- 229940035936 ubiquinone Drugs 0.000 description 12
- -1 silicon ion Chemical class 0.000 description 11
- 244000061458 Solanum melongena Species 0.000 description 10
- 235000002597 Solanum melongena Nutrition 0.000 description 10
- 238000012360 testing method Methods 0.000 description 10
- 238000009395 breeding Methods 0.000 description 9
- 230000001488 breeding effect Effects 0.000 description 9
- 230000000694 effects Effects 0.000 description 9
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- 230000012010 growth Effects 0.000 description 9
- FPIPGXGPPPQFEQ-UHFFFAOYSA-N 13-cis retinol Natural products OCC=C(C)C=CC=C(C)C=CC1=C(C)CCCC1(C)C FPIPGXGPPPQFEQ-UHFFFAOYSA-N 0.000 description 8
- VTYYLEPIZMXCLO-UHFFFAOYSA-L Calcium carbonate Chemical compound [Ca+2].[O-]C([O-])=O VTYYLEPIZMXCLO-UHFFFAOYSA-L 0.000 description 8
- FPIPGXGPPPQFEQ-BOOMUCAASA-N Vitamin A Natural products OC/C=C(/C)\C=C\C=C(\C)/C=C/C1=C(C)CCCC1(C)C FPIPGXGPPPQFEQ-BOOMUCAASA-N 0.000 description 8
- FPIPGXGPPPQFEQ-OVSJKPMPSA-N all-trans-retinol Chemical compound OC\C=C(/C)\C=C\C=C(/C)\C=C\C1=C(C)CCCC1(C)C FPIPGXGPPPQFEQ-OVSJKPMPSA-N 0.000 description 8
- GVPFVAHMJGGAJG-UHFFFAOYSA-L cobalt dichloride Chemical compound [Cl-].[Cl-].[Co+2] GVPFVAHMJGGAJG-UHFFFAOYSA-L 0.000 description 8
- 239000007788 liquid Substances 0.000 description 8
- 235000019155 vitamin A Nutrition 0.000 description 8
- 239000011719 vitamin A Substances 0.000 description 8
- 229940045997 vitamin a Drugs 0.000 description 8
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 7
- 239000007791 liquid phase Substances 0.000 description 7
- 229910052760 oxygen Inorganic materials 0.000 description 7
- 239000001301 oxygen Substances 0.000 description 7
- 238000003786 synthesis reaction Methods 0.000 description 7
- 229940099596 manganese sulfate Drugs 0.000 description 6
- 239000011702 manganese sulphate Substances 0.000 description 6
- 235000007079 manganese sulphate Nutrition 0.000 description 6
- SQQMAOCOWKFBNP-UHFFFAOYSA-L manganese(II) sulfate Chemical compound [Mn+2].[O-]S([O-])(=O)=O SQQMAOCOWKFBNP-UHFFFAOYSA-L 0.000 description 6
- 229930003448 Vitamin K Natural products 0.000 description 5
- 230000027721 electron transport chain Effects 0.000 description 5
- 238000002474 experimental method Methods 0.000 description 5
- SHUZOJHMOBOZST-UHFFFAOYSA-N phylloquinone Natural products CC(C)CCCCC(C)CCC(C)CCCC(=CCC1=C(C)C(=O)c2ccccc2C1=O)C SHUZOJHMOBOZST-UHFFFAOYSA-N 0.000 description 5
- 235000019168 vitamin K Nutrition 0.000 description 5
- 239000011712 vitamin K Substances 0.000 description 5
- 150000003721 vitamin K derivatives Chemical class 0.000 description 5
- 229940046010 vitamin k Drugs 0.000 description 5
- 229920001817 Agar Polymers 0.000 description 4
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 4
- 241000191043 Rhodobacter sphaeroides Species 0.000 description 4
- 240000008042 Zea mays Species 0.000 description 4
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 description 4
- 235000002017 Zea mays subsp mays Nutrition 0.000 description 4
- 239000008272 agar Substances 0.000 description 4
- 229910000019 calcium carbonate Inorganic materials 0.000 description 4
- 235000005822 corn Nutrition 0.000 description 4
- 238000011081 inoculation Methods 0.000 description 4
- 239000000843 powder Substances 0.000 description 4
- 238000004321 preservation Methods 0.000 description 4
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 3
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 3
- 230000008901 benefit Effects 0.000 description 3
- 239000005515 coenzyme Substances 0.000 description 3
- 238000010586 diagram Methods 0.000 description 3
- 244000005700 microbiome Species 0.000 description 3
- 239000000203 mixture Substances 0.000 description 3
- 150000003839 salts Chemical class 0.000 description 3
- 229940088594 vitamin Drugs 0.000 description 3
- 229930003231 vitamin Natural products 0.000 description 3
- 235000013343 vitamin Nutrition 0.000 description 3
- 239000011782 vitamin Substances 0.000 description 3
- 150000003722 vitamin derivatives Chemical class 0.000 description 3
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 2
- 102000018832 Cytochromes Human genes 0.000 description 2
- 108010052832 Cytochromes Proteins 0.000 description 2
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 2
- 241000196324 Embryophyta Species 0.000 description 2
- 102000001554 Hemoglobins Human genes 0.000 description 2
- 108010054147 Hemoglobins Proteins 0.000 description 2
- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 description 2
- 102000005298 Iron-Sulfur Proteins Human genes 0.000 description 2
- 108010081409 Iron-Sulfur Proteins Proteins 0.000 description 2
- 241001465754 Metazoa Species 0.000 description 2
- 244000061176 Nicotiana tabacum Species 0.000 description 2
- 235000002637 Nicotiana tabacum Nutrition 0.000 description 2
- 241001052560 Thallis Species 0.000 description 2
- 239000003963 antioxidant agent Substances 0.000 description 2
- 230000003078 antioxidant effect Effects 0.000 description 2
- 235000006708 antioxidants Nutrition 0.000 description 2
- 229910052799 carbon Inorganic materials 0.000 description 2
- 230000010261 cell growth Effects 0.000 description 2
- 235000019441 ethanol Nutrition 0.000 description 2
- 238000011049 filling Methods 0.000 description 2
- 239000001257 hydrogen Substances 0.000 description 2
- 229910052739 hydrogen Inorganic materials 0.000 description 2
- 238000009776 industrial production Methods 0.000 description 2
- 239000002054 inoculum Substances 0.000 description 2
- 238000011068 loading method Methods 0.000 description 2
- 230000000813 microbial effect Effects 0.000 description 2
- LPUQAYUQRXPFSQ-DFWYDOINSA-M monosodium L-glutamate Chemical compound [Na+].[O-]C(=O)[C@@H](N)CCC(O)=O LPUQAYUQRXPFSQ-DFWYDOINSA-M 0.000 description 2
- 235000013923 monosodium glutamate Nutrition 0.000 description 2
- 239000004223 monosodium glutamate Substances 0.000 description 2
- 229910052757 nitrogen Inorganic materials 0.000 description 2
- 239000012071 phase Substances 0.000 description 2
- 239000011734 sodium Substances 0.000 description 2
- 229910052708 sodium Inorganic materials 0.000 description 2
- 230000007480 spreading Effects 0.000 description 2
- 238000003892 spreading Methods 0.000 description 2
- 239000008223 sterile water Substances 0.000 description 2
- 238000004659 sterilization and disinfection Methods 0.000 description 2
- 230000004083 survival effect Effects 0.000 description 2
- 239000000725 suspension Substances 0.000 description 2
- 238000012549 training Methods 0.000 description 2
- 238000005406 washing Methods 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- 102000005606 Activins Human genes 0.000 description 1
- 108010059616 Activins Proteins 0.000 description 1
- 241000589158 Agrobacterium Species 0.000 description 1
- 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 1
- 208000024172 Cardiovascular disease Diseases 0.000 description 1
- ZAMOUSCENKQFHK-UHFFFAOYSA-N Chlorine atom Chemical compound [Cl] ZAMOUSCENKQFHK-UHFFFAOYSA-N 0.000 description 1
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 description 1
- NBIIXXVUZAFLBC-UHFFFAOYSA-L Phosphate ion(2-) Chemical compound OP([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-L 0.000 description 1
- ZLMJMSJWJFRBEC-UHFFFAOYSA-N Potassium Chemical compound [K] ZLMJMSJWJFRBEC-UHFFFAOYSA-N 0.000 description 1
- 241000316848 Rhodococcus <scale insect> Species 0.000 description 1
- 238000009825 accumulation Methods 0.000 description 1
- 230000004913 activation Effects 0.000 description 1
- 239000000488 activin Substances 0.000 description 1
- 238000005937 allylation reaction Methods 0.000 description 1
- 230000003698 anagen phase Effects 0.000 description 1
- 230000003712 anti-aging effect Effects 0.000 description 1
- 239000003054 catalyst Substances 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 238000005660 chlorination reaction Methods 0.000 description 1
- 239000000460 chlorine Substances 0.000 description 1
- 229910052801 chlorine Inorganic materials 0.000 description 1
- 230000000052 comparative effect Effects 0.000 description 1
- 230000002860 competitive effect Effects 0.000 description 1
- 238000006482 condensation reaction Methods 0.000 description 1
- 239000000287 crude extract Substances 0.000 description 1
- 238000012136 culture method Methods 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000018109 developmental process Effects 0.000 description 1
- 230000002708 enhancing effect Effects 0.000 description 1
- 125000001495 ethyl group Chemical group [H]C([H])([H])C([H])([H])* 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 235000013373 food additive Nutrition 0.000 description 1
- 239000002778 food additive Substances 0.000 description 1
- 239000003966 growth inhibitor Substances 0.000 description 1
- 125000004435 hydrogen atom Chemical group [H]* 0.000 description 1
- 230000036039 immunity Effects 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 239000003112 inhibitor Substances 0.000 description 1
- 239000012533 medium component Substances 0.000 description 1
- 229910052751 metal Chemical class 0.000 description 1
- 239000002184 metal Chemical class 0.000 description 1
- 230000002906 microbiologic effect Effects 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 229910000402 monopotassium phosphate Inorganic materials 0.000 description 1
- 235000019796 monopotassium phosphate Nutrition 0.000 description 1
- 238000002703 mutagenesis Methods 0.000 description 1
- 231100000350 mutagenesis Toxicity 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 235000016709 nutrition Nutrition 0.000 description 1
- 230000035764 nutrition Effects 0.000 description 1
- 238000005457 optimization Methods 0.000 description 1
- 239000003208 petroleum Substances 0.000 description 1
- PJNZPQUBCPKICU-UHFFFAOYSA-N phosphoric acid;potassium Chemical compound [K].OP(O)(O)=O PJNZPQUBCPKICU-UHFFFAOYSA-N 0.000 description 1
- 230000001766 physiological effect Effects 0.000 description 1
- 238000011020 pilot scale process Methods 0.000 description 1
- 239000011591 potassium Substances 0.000 description 1
- 229910052700 potassium Inorganic materials 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 230000002265 prevention Effects 0.000 description 1
- 150000004053 quinones Chemical class 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 238000010188 recombinant method Methods 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 229910052710 silicon Inorganic materials 0.000 description 1
- 239000010703 silicon Substances 0.000 description 1
- 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 1
- 238000003756 stirring Methods 0.000 description 1
- 238000013517 stratification Methods 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 238000004114 suspension culture Methods 0.000 description 1
- 230000002194 synthesizing effect Effects 0.000 description 1
- ITMCEJHCFYSIIV-UHFFFAOYSA-N triflic acid Chemical class OS(=O)(=O)C(F)(F)F ITMCEJHCFYSIIV-UHFFFAOYSA-N 0.000 description 1
- 125000001655 ubiquinone group Chemical group 0.000 description 1
- 238000010200 validation analysis Methods 0.000 description 1
- 235000013311 vegetables Nutrition 0.000 description 1
Classifications
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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
-
- 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
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- 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)
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- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
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Abstract
A kind of raising ubiquinone10The production method of industrial output, it include: that for the bacterial strain of CGMCC No.5997, CGMCC No.5998 or CGMCC No.5999, successively screening obtains fermentation seed after recovery and seed culture medium spread cultivation by deposit number, it is seeded to after gained fermentation seed is spread cultivation in fermentation medium and carries out fermented and cultured, obtain ubiquinone10, Fe in seed culture medium2+Concentration be 0.1~0.5mol/L.The present invention solves existing fermentation method production ubiquinone10The not high technical problem of industrial output.
Description
Technical field
The present invention relates to microorganism fields, and in particular to a kind of to promote hydrogenlike silicon ion Rhodobacter
Sphaeroides produces ubiquinone10The peculiar cultural method of strain activity and yield.
Background technique
Ubiquinone10(Coenzyme Q10) it is also known as ubiquinone (Ubiquinone, abridge UQ), it is a kind of to be present in nature
Fat-soluble quinones is hydrogen carrier important on electron transport chain in organism.Energy is participated in into the cell in human body
Manufacture and activation, are that prevention of arterial hardens to form most effective anti-oxidant composition.Ubiquinone10It can human activin cell and cell energy
The nutrition of amount has the function of to improve the immunity of the human body, enhances anti-oxidant, anti-aging and enhancing human activity etc., medically extensively
It is general to be used for disease of cardiovascular system, it is used for nutrient and healthcare products and food additives extensively both at home and abroad.
Ubiquinone10Preparation method there are mainly three types of: animal vegetable tissue cytapheresis, chemical synthesis and microorganism hair
Ferment method.Since production efficiency is lower, technique falls behind animal and plant cells extraction method, does not have to substantially in industrialized production at present.
For example, the Chinese invention patent application of Publication No. CN 102391093A discloses one kind extracts ubiquinone from tobacco10Side
Method, process flow are: by the tobacco sale separated with culture solution or tobacco suspension culture cell tissue block and 75~
The mixing of 85% ethyl alcohol-petroleum ether solution homogenate 5~10 minutes, above-mentioned homogenate mixtures filtering after homogenate, stratification, on
Up to ubiquinone after layer solution concentration10Crude extract.
Ubiquinone is produced with chemical synthesis10, for example, the Chinese invention patent application of Publication No. CN 1931818 discloses
A kind of synthesizing coenzyme Q10Method.This method is with ubiquinone0Quinhydrones and content are not less than 95% isodeca-deca-isoprene-yl
Alcohol is raw material, after being dehydrated, through trifluoromethanesulfonic acid and its metal salt catalyst in oxygen-free environment, and condensation reaction under room temperature,
Obtain ubiquinone10Quinhydrones,.Chemical synthesis yield is relatively low, and the allylation reagents made from solanesol are suitable, anteiso-s
The mixture of structure body, activity is not strong, needs to separate, therefore the application of this method is restricted.
Microbe fermentation method is the hot spot of global development in recent years, it is considered to be most promising production method.Using micro-
Biological fermentation process produces ubiquinone10, either in terms of the quality of product and safety, have biggish competitive advantage, be suitable for
Large-scale industrial production.
Such as the Chinese invention patent application of Publication No. CN 101024849 discloses a kind of ubiquinone10Photosynthetic bacteria
Fermentation method: the building of DNA recombinant technique, screening high yield ubiquinone are utilized10Photosynthetic bacterium strain, meanwhile, using chemical substance or ultraviolet
Line mutagenesis screens high yield ubiquinone using carotenogenesis inhibitor and growth inhibitor10Photosynthetic bacteria mutant, it is real
Existing high yield ubiquinone10The breeding and cultivation of bacterial strain.The Chinese invention patent application of Publication No. CN 1884562 discloses a kind of life
Produce ubiquinone10Method, produced using fermentation process, ferment used by strain be Agrobacterium LQ10。
All there is ubiquinone when existing microbe fermentation method is in particular by engineering bacteria10The not high problem of industrial output.
Summary of the invention
The present invention provides a kind of raising ubiquinone10The production method of industrial output solves existing fermentation method production coenzyme
Q10The not high technical problem of industrial output.
A kind of raising ubiquinone10The production method of industrial output, comprising: by deposit number be CGMCC No.5997, CGMCC
Successively screening obtains fermentation seed to the bacterial strain of No.5998 or CGMCC No.5999 after recovering and seed culture medium spreads cultivation, by institute
It obtains to be seeded in fermentation medium after fermentation seed spreads cultivation and carries out fermented and cultured, obtain ubiquinone10, Fe in seed culture medium2+'s
Concentration is 0. 1~0.5mol/L.
The bacterial strain that deposit number is CGMCC No.5997 is the red ball bacterium of class, Latin name rhodobacter
Sphaeroides is named as NHU-ZUB bacterial strain.
The bacterial strain of CGMCC No.5998 is the red ball bacterium of class, and Latin name is rhodobacter sphaeroides,
It is named as NHU-ZDD bacterial strain.
The bacterial strain of CGMCC No.5999 is the red ball bacterium of class, and Latin name is rhodobacter sphaeroides,
It is named as NHU-ZAA bacterial strain.
The above bacterial strain be applicant China Committee for Culture Collection of Microorganisms's common micro-organisms center's preservation simultaneously
The bacterial strain of survival.
In microbial fermentation production, the excellent of strain is the key that fermenting and producing yield height by the gross, is accessed in bacterial strain
Before big tank, selecting outstanding bacterial strain is the guarantee of high yield, and bacterial screening is to improve strain quality, stablize the important of fermentation level
Means.And suitable growing environment is the guarantee of bacterial strain fast-growth, gives the demand of bacterial strain abundance, bacterial strain could be allowed not by short
The limitation of plate effect is farthest production service.
Ubiquinone10It is hydrogen carrier important on electron transport chain in organism, thus enhances electron transport chain intensity and be conducive to
Ubiquinone10Accumulation.Wherein, iron ion is iron-sulfur protein in electron transport chain, hemoglobin in cytochromes and organism
Important prothetic group.
It is found in researcher's research of the present invention, in current published report, iron ion is used as the micro rush of culture medium
Used into agent, and it is rarely seen screen the strain for being applicable in industrial production and using using adding iron concentration in culture medium, it is especially suitable
Close the strain after being genetically engineered.
The present invention is keeping ubiquinone10Under the premise of original quality of fermenting, according to ubiquinone10The physiological property of bacterium is produced, is led to
A large amount of flask process optimization experiment is crossed, to effectively improve strain activity and ubiquinone10For the purpose of yield, establishes one kind and effectively may be used
Capable culture screening technique, having found a kind of can effectively improve ubiquinone10Produce the fermentation training of bacterial strain microbial activity and production yields
The method of supporting, and expansion is realized in production, effectively increase production yields.
Preferably, Fe in the seed culture medium2+Concentration be 0.1~0.3mol/L；It is further preferred that seed is trained
Support Fe in base2+Concentration be 0.3mol/L.
It is further preferred that the condition of culture to spread cultivation in seed culture medium are as follows: 30~32 DEG C are protected from light culture 22~for 24 hours.It goes out
121 DEG C of bacterium temperature, sterilization time 20min.
Preferably, the recovery successively carries out in plating medium and slant medium, plating medium and seed training
Support Fe in base2+Concentration be 0.1~0.5mol/L.
It is further preferred that Fe in the plating medium2+Concentration be 0.1~0.3mol/L；It is further preferred that
Fe in the plating medium2+Concentration be 0.3mol/L.
It is further preferred that Fe in the slant medium2+Concentration be 0.1~0.3mol/L；It is further preferred that
Fe in the slant medium2+Concentration be 0.3mol/L.
It is further preferred that the condition of culture in slant medium is 30~32 DEG C and is protected from light 60~80h of culture.Sterilizing temperature
121 DEG C of degree, sterilization time 20min.
Preferably, Fe2+It is added to culture at least one of iron chloride, ferrous sulfate and ironic citrate compound form
In base.
It is highly preferred that Fe in plating medium, slant medium and seed culture medium2+Concentration be 0.1~
0.3mol/L；Most preferably, Fe in plating medium, slant medium and seed culture medium2+Concentration be 0.3mol/L.
Plating medium and slant medium used in the present invention are that class Rhodococcus sp often uses culture medium, including carbon source, nitrogen source, nothing
The conventional ingredients such as machine salt, vitamin.
Preferably, plating medium (every 100mL) ingredient are as follows: glucose 0.3g, yeast extract 0.8g, cobalt chloride
0.003g, sodium chloride 0.2g, manganese sulfate 0.0001g, dipotassium hydrogen phosphate 0.13g, magnesium sulfate 0.025g, vitaminB10 .1ug, dimension
Raw element K 0.1ug, vitamin A 0.15ug, agar powder 1.5g, then add the Fe of 0.1~0.5mol/L2+.PH is 7.2, and sterilize item
Part can be 118 DEG C, 10min.
The ingredient (every 100mL) of slant medium are as follows: glucose 0.3g, yeast extract 0.8g, cobalt chloride 0.003g, chlorine
Change sodium 0.2g, manganese sulfate 0.0001g, dipotassium hydrogen phosphate 0.13g, magnesium sulfate 0.025g, vitaminB10 .1ug, vitamin K
0.1ug, vitamin A 0.15ug, agar powder 1.2g；The Fe of 0.1~0.5mol/L is added again2+.PH is 7.2, and sterilising conditions can
Think 118 DEG C, 10min.
Seed culture medium (every 100mL) ingredient are as follows: ammonium sulfate 0.25g, corn pulp 0.05g, glucose 0.3g, yeast extract
Object 0.14g, cobalt chloride 0.003g, sodium chloride 0.2g, manganese sulfate 0.0001g, dipotassium hydrogen phosphate 0.05g, potassium dihydrogen phosphate
0.05g, magnesium sulfate 0.1g, calcium carbonate 0.8g, vitaminB10 .1ug, vitamin K 0.1ug, vitamin A 0.15ug, then add
Add the Fe of 0.1~0.5mol/L2+.PH is 7.2, and sterilising conditions can be 118 DEG C, 10min.
Bacterial screening process of the invention specifically includes the following steps:
1) primary dcreening operation (recovery) of strain: spot inoculation small test tube slant medium is chosen using plate single colonie, when cultivating one section
Between after inoculation fermentation shake flask test sample screening；
2) secondary screening (spreading cultivation) of strain: the result obtained from primary dcreening operation selects outstanding bacterial strain, and cryovial connects small female bottle and then connects
Fermentation flask test sample secondary screening；
3) preservation of strain: the bacterial strain that secondary screening obtains is carried out applying eggplant bottle inclined-plane, adds glycerol stocks after culture；
4) production and application of bacterial strain: the big female bottle of the outstanding strain inoculated that secondary screening is obtained, it is female as one grade fermemtation after culture
Kind, it is seeded in fermentor and cultivates after spreading cultivation.
Fermentation flask incubation time is protected from light culture 48h for 31 DEG C in screening step (1) and (2), 121 DEG C of sterilising temp, sterilizes
Time 20min.
In screening step (3), the eggplant bottle inclined-plane culture time is 31 DEG C and is protected from light culture 3 days, 121 DEG C of sterilising temp, sterilizes
Time 20min.
It is that bacterial strain uses therefor of the present invention often uses fermentation medium that fermentation medium used in potency is detected in above-mentioned screening process, is wrapped
Include the conventional ingredients such as carbon source, nitrogen source, inorganic salts, vitamin.
Preferably, the fermentation medium (every 100mL): ammonium sulfate 0.3g, glucose 4g, monosodium glutamate 0.3g, cobalt chloride
0.005g, sodium chloride 0.28g, calcium carbonate 0.6g, dipotassium hydrogen phosphate 0.15g, magnesium sulfate 0.63g, ferrous sulfate 0.12g, corn
0.4g vitaminB10 .1ug, vitamin K 0.1ug, vitamin A 0.15ug, pH 7.2 are starched, sterilising conditions can be 118 DEG C,
10min。
Detect fermentation culture conditions when potency are as follows: it is small to be protected from light culture 48~96 in 26~34 DEG C, 200rpm~250rpm
When, it extracts fermentation liquid and detects potency.The purpose of stirring is supplied oxygen for fermentation process, adjusts revolving speed to reach adjustment for oxygen concentration
Purpose can be further improved ubiquinone in conjunction with the oxygen supply in fermentation process on the basis of bacterial strain screening of the present invention10Yield.
Spread cultivation process of the strain screened in the present invention when producing and using is as follows:
(1) inclined-plane culture: the glycerol tube of preservation is connected on eggplant type bottle inclined-plane, is cultivated 3 days at 31 DEG C.
(2) with the cultured inclined-plane of sterile water washing, 10 seed culture: are made8~109The bacteria suspension of a cells/ml,
In the seed bottle for pipetting 10ml to loading amount 500ml/1000ml, 30 DEG C, cultivate 22-26 hours under conditions of 180~250rpm
To seed liquor.
Seed liquor obtained by above-mentioned steps (2) can directly be seeded to fermentation cylinder for fermentation culture, inoculum concentration 10%, in fermentation
The ingredient of fermentation medium is preferred are as follows: the fermentation medium components are as follows: glucose 1.8-2.5g/L, yeast extract 7.5~
8.5g/L, 2.5~3g/L of sodium chloride, 0.5~0.8g/L of dipotassium hydrogen phosphate, 0.2~0.3g/L of magnesium sulfate, ammonium sulfate 2.5~
3.5g/L, pH are 6.5~7.0.
Further preferably are as follows: glucose 1.8-2.5g/L, yeast extract 8g/L, sodium chloride 2.8g/L, dipotassium hydrogen phosphate 0.6g/
L, magnesium sulfate 0.25g/L, ammonium sulfate 3g/L, pH 6.8.Condition of culture in fermentor are as follows: 29-33 DEG C of cultivation temperature, tank internal pressure
0.03~0.05Mpa of power, 5~6L/min of air mass flow, speed of agitator 500-700rpm.
Glucose is continuously added according to thalli growth situation adjusting process parameter and beginning in fermentation process.
The present invention is for the first time in ubiquinone10Distinctive screening and culturing method is used in entire fermenting and producing process, and use adds
Add the different compounds of the iron of suitable concentration to improve the various culture mediums of strain activity and yield, preferably adds iron concentration
0.1~0.5mol/L.Iron is that cell grows necessary element, and the appropriate iron ion that adds can promote the normal growth of cell.Iron from
Son be iron-sulfur protein in electron transport chain, in cytochromes and organism hemoglobin important prothetic group.In previous production,
It is inadequate to the concern of iron ion, it often only adds the compound of Single Iron on a small quantity in the medium, limits the quick of cell
Growth, the present invention are added the compound of the iron of variety classes and proportion in the medium by lot of experiment validation, can improved
Strain growth activity and production ubiquinone10Ability.
It can be seen that the present invention has following advantages compared to pervious zymotechnique: 1. the growth vigor of bacterium cell is strong,
Tolerance is strong, is easy production in the fermenter；2. physiological status is stablized, stable production capacity is kept；3. producing strain is stablized
And yield can be slowly improved, guarantee production capacity in the survival of the fittest.
The present invention is for the first time in ubiquinone10Come in fermenting and producing using the culture medium of the compound state of addition suitable concentration difference iron
The fermentation culture method for being screened and being cultivated while carried out, preferably addition 0.1~0.5mol/L of iron concentration, and be applied to
In pilot scale and big examination fermentation, ideal fermentation results are obtained.
Detailed description of the invention
Fig. 1 is to add the compound of different iron to ubiquinone10Synthesis influences comparison diagram.
Fig. 2 is different ferrous sulfate concentration to ubiquinone10Synthesis influences comparison diagram.
Fig. 3 is to add iron ion in different rotating speeds to ubiquinone10Synthesis influences comparison diagram.
Specific embodiment
(1) plating medium basic ingredient (every 100mL): glucose 0.3g, yeast extract 0.8g, cobalt chloride
0.003g, sodium chloride 0.2g, manganese sulfate 0.0001g, dipotassium hydrogen phosphate 0.13g, magnesium sulfate 0.025g, vitaminB10 .1ug, dimension
Raw element K 0.1ug, vitamin A 0.15ug, agar powder 1.5g, pH 7.2, sterilising conditions can be 118 DEG C, 10min.
(2) slant medium basic ingredient (every 100mL): glucose 0.3g, yeast extract 0.8g, cobalt chloride
0.003g, sodium chloride 0.2g, manganese sulfate 0.0001g, dipotassium hydrogen phosphate 0.13g, magnesium sulfate 0.025g, vitaminB10 .1ug, dimension
Raw element K 0.1ug, vitamin A 0.15ug, agar powder 1.2g, pH 7.2, sterilising conditions can be 118 DEG C, 10min.
(3) seed culture medium basic ingredient (every 100mL): ammonium sulfate 0.25g, corn pulp 0.05g, glucose 0.3g, ferment
Female extract 0.14g, cobalt chloride 0.003g, sodium chloride 0.2g, manganese sulfate 0.0001g, dipotassium hydrogen phosphate 0.05g, biphosphate
Potassium 0.05g, magnesium sulfate 0.1g, calcium carbonate 0.8g, vitaminB10 .1ug, vitamin K 0.1ug, vitamin A 0.15ug, pH are
7.2, sterilising conditions can be 118 DEG C, 10min.
(4) fermentation medium (every 100mL): ammonium sulfate 0.3g, glucose 4g, monosodium glutamate 0.3g, cobalt chloride 0.005g, chlorination
Sodium 0.28g, calcium carbonate 0.6g, dipotassium hydrogen phosphate 0.15g, magnesium sulfate 0.63g, corn pulp 0.4g, FeSO40.12g, vitamin
B10.1ug, vitamin K 0.1ug, vitamin A 0.15ug, pH 7.2, sterilising conditions can be 118 DEG C, 10min.
(5) fermentation tank culture medium: glucose 1.8-2.5g/L, yeast extract 8g/L, sodium chloride 2.8g/L, dipotassium hydrogen phosphate
0.6g/L, magnesium sulfate 0.25g/L, ammonium sulfate 3g/L, pH 6.8, sterilising conditions can be 118 DEG C, 10min.
Fermentation liquid purpose product ubiquinone is given below10Using liquid phase testing conditions:
Chromatographic column: C18,4.6*150mm, 5um；
Detection wavelength: 275nm；
Mobile phase: ethyl alcohol: methanol=1:1；
Flow velocity: 1.5ml/min；
Column temperature: 35 DEG C.
Embodiment 1: the different compounds of iron are added and do not add shaking flask ubiquinone10The comparison of fermentation.
Shaking flask primary dcreening operation breeding fermentation process: plate single colonie → small test tube inclined-plane → fermentation flask → liquid phase detects Q10Potency →
Glycerol tube.Detailed process is as follows:
7 days or so single colonies are cultivated in selection on plate, and correspondence is chosen in small test tube inclined-plane, after 31 DEG C are protected from light culture 3 days
It is washed with 4mL, draws the 250mL shaking flask of 1mL bacterium solution to dress 50ml fermentation medium, 31 DEG C, 230r/min is protected from light culture 48h,
It extracts fermentation liquid and detects potency, remaining bacterium solution refrigerator saves stand-by.The outstanding bacterial strain bacterium solution addition same volume of result will be measured
40% glycerol conservation.
Shaking flask secondary screening breeding fermentation process: primary dcreening operation glycerol tube → small female bottle → fermentation flask → liquid phase detects Q10Potency → eggplant
Sub- bottle inclined-plane spreads cultivation and is used to produce.Detailed process is as follows:
It chooses primary dcreening operation and obtains the 250mL shaking flask of glycerol tube access dress 50ml seed culture medium, 31 DEG C, 230r/min is protected from light
Cultivate 23h, draw 1mL bacterium solution to fill 50ml fermentation medium 250mL shaking flask, 31 DEG C, 230r/min be protected from light culture 48h,
96h extracts fermentation liquid respectively and detects potency.It selects outstanding bacterial strain cryovial to apply eggplant bottle inclined-plane to spread cultivation, the bacterial strain to spread cultivation out connects
Seed bottle application mass production.
The compound of 0.1mol/L iron is added in plate, inclined-plane and seed culture medium in the shake flat experiment result that do not add
Comparison such as table 1, table 1: adds the different compounds of iron and the fermentation results not added.
The result shows that: the compound that iron is added in culture medium embodies following advantages:
1. single colonie growth is fast on plate, bacterium colony color is bright and new full；
2. the growth of bacterial strain Individual Size is fast in seed bottle, form is mellow and full, and whole bacterium is dense higher；
3. strain growth is very fast in fermentation flask, potency is higher.It is more excellent to add ferrous sulfate.
Embodiment 2: the ferrous sulfate concentration shaking flask ubiquinone of different gradients is added in culture medium10The comparison of fermentation.
Shaking flask primary dcreening operation breeding fermentation process: plate single colonie → small test tube inclined-plane → fermentation flask → liquid phase detects Q10Potency →
Glycerol tube.Detailed process is as follows:
7 days or so single colonies are cultivated in selection on plate, and correspondence is chosen in small test tube inclined-plane, after 31 DEG C are protected from light culture 3 days
It is washed down with 4mL, draws 1mL bacterium solution to the 250mL shaking flask for filling 50ml fermentation medium, 31 DEG C, 230r/min is protected from light culture
48h extracts fermentation liquid and detects potency, and remaining bacterium solution refrigerator saves stand-by.The outstanding bacterial strain bacterium solution addition same volume of result will be measured
40% glycerol conservation of product.
Shaking flask secondary screening breeding fermentation process: primary dcreening operation glycerol tube → small female bottle → fermentation flask → liquid phase detects Q10Potency → eggplant
Sub- bottle inclined-plane spreads cultivation and is used to produce.Detailed process is as follows:
It chooses primary dcreening operation and obtains the 250mL shaking flask of glycerol tube access dress 50ml seed culture medium, 31 DEG C, 230r/min is protected from light
Cultivate 23h, draw 1mL bacterium solution to fill 50ml fermentation medium 250mL shaking flask, 31 DEG C, 230r/min be protected from light culture 48h,
96h extracts fermentation liquid respectively and detects potency.It selects outstanding bacterial strain cryovial to apply eggplant bottle inclined-plane to spread cultivation, the bacterial strain to spread cultivation out connects
Seed bottle application mass production.
The ferrous sulfate concentration of different gradients is added in plate, inclined-plane and seed culture medium, the strain screened exists
Shake flat experiment Comparative result such as table 2:
Table 2: different gradient ferrous sulfate fermenting experiment comparing results are added:
The result shows that after adding the 0.1~0.5mol/L breeding of ferrous sulfate concentration in plate and seed culture medium, effect
It is particularly evident.
Embodiment 3: the shake flask fermentation Co-Q10 after addition iron ion under different oxygen supply conditions compares.
Shaking flask primary dcreening operation breeding fermentation process: plate single colonie → small test tube inclined-plane → fermentation flask → liquid phase detects Q10Potency →
Glycerol tube.Detailed process is as follows:
7 days or so single colonies are cultivated in selection on plate, and correspondence is chosen in small test tube inclined-plane, after 31 DEG C are protected from light culture 3 days
It is washed down with 4mL, draws 1mL bacterium solution to the 250mL shaking flask for filling 50ml fermentation medium, 31 DEG C, 230r/min is protected from light culture
48h extracts fermentation liquid and detects potency, and remaining bacterium solution refrigerator saves stand-by.The outstanding bacterial strain bacterium solution addition same volume of result will be measured
40% glycerol conservation of product.
Shaking flask secondary screening breeding fermentation process: primary dcreening operation glycerol tube → small female bottle → fermentation flask → liquid phase detects Q10Potency → eggplant
Sub- bottle inclined-plane spreads cultivation and is used to produce.Detailed process is as follows:
It chooses primary dcreening operation and obtains the 250mL shaking flask of glycerol tube access dress 50ml seed culture medium, 31 DEG C, 230r/min is protected from light
Cultivate 23h, draw 1mL bacterium solution to fill 50ml fermentation medium 250mL shaking flask, 31 DEG C, 230r/min be protected from light culture 48h,
96h extracts fermentation liquid respectively and detects potency.It selects outstanding bacterial strain cryovial to apply eggplant bottle inclined-plane to spread cultivation, the bacterial strain to spread cultivation out connects
Seed bottle application mass production.
By adding the breeding of 0.3mol/L ferrous sulfate plate and being added to the seed culture of 0.3mol/L ferrous sulfate
Base spread cultivation after seed, be connected in fermentation shake flask, under equal conditions, under different rotating speeds, coenzyme under more different oxygen supply conditions
Q10Ferment effect.
Table 3: shaking flask ubiquinone under the conditions of different rotating speeds10Fermentation results
The result shows that increasing under oxygen supply condition, addition iron ion can more preferably promote ubiquinone10Synthesis.
Embodiment 4: the ferment tank Co-Q10 result after addition iron ion in different strains compares.
Fermentor investigates process: glycerol tube → inclined-plane culture → mother's bottle seed → fermentor, detailed process are as follows；
(1) inclined-plane culture: the glycerol tube of preservation is connected on eggplant type bottle inclined-plane, is cultivated 3 days at 31 DEG C.
(2) with the cultured inclined-plane of sterile water washing, 10 seed culture: are made8~109The bacteria suspension of a cells/ml,
In the seed bottle for pipetting 10ml to loading amount 500ml/1000ml, 30 DEG C, cultivate 22-26 hours under conditions of 180~250rpm
To seed liquor.
(3) seed liquor obtained by step (2) fermented and cultured: is inoculated into 10 liters of fermentors, inoculum concentration 10%, cultivation temperature
29-33 DEG C, 0.03~0.05Mpa of pressure inside the tank, control by stages strategy is taken in oxygen supply, initial speed of agitator after inoculation
500rpm, air mass flow 6L/min, after inoculation, as growth lag phase terminates, thallus starts fast-growth and enters logarithmic growth
Phase, OUR rapid increase, 24 hours, and maintain 30-50mmol/Lh, revolving speed 500-700rpm, residual glucose control in tank
System is cultivated 110 hours or so between 0.5-2.0%.In the process according to thalli growth situation adjusting process parameter and beginning
Continuously add glucose.
Compare inclined-plane, plant in Combined inkbottle and add and do not add the fermentation results situation between different strains:
The result shows that increase iron ion in slant medium and female bottle culture medium, and under identical fermentation condition, ubiquinone10
Yield has the growth of certain amplitude.
Claims (7)
1. a kind of raising ubiquinone10The production method of industrial output, comprising: by deposit number be CGMCC No.5997, CGMCC
Successively screening obtains fermentation seed to the bacterial strain of No.5998 or CGMCC No.5999 after recovering and seed culture medium spreads cultivation, by institute
It obtains to be seeded in fermentation medium after fermentation seed spreads cultivation and carries out fermented and cultured, obtain ubiquinone10, which is characterized in that the recovery
Successively carried out in plating medium and slant medium；Fe in plating medium and seed culture medium2+Concentration be 0.1~
0.5mol/L；Fe in slant medium2+Concentration be 0.1~0.3mol/L；
The ingredient of the fermentation medium are as follows: glucose 1.8-2.5g/L, 7.5~8.5g/L of yeast extract, 2.5~3g/ of sodium chloride
L, 0.5~0.8g/L of dipotassium hydrogen phosphate, 0.2~0.3g/L of magnesium sulfate, ammonium sulfate 2.5~3.5g/L, pH are 6.5~7.0.
2. production method according to claim 1, which is characterized in that Fe in the seed culture medium2+Concentration be 0.1~
0.3mol/L。
3. production method according to claim 1, which is characterized in that the condition of culture to spread cultivation in seed culture medium are as follows: 30
~32 DEG C are protected from light culture 22~for 24 hours.
4. production method according to claim 1, which is characterized in that Fe in the plating medium2+Concentration be 0.1~
0.3mol/L。
5. production method according to claim 1, which is characterized in that the condition of culture in slant medium is 30~32 DEG C
It is protected from light 60~80h of culture.
6. production method according to claim 1, which is characterized in that Fe2+In iron chloride, ferrous sulfate and ironic citrate
At least one compound form be added in culture medium.
7. production method according to claim 1, which is characterized in that cultivation temperature 29-33 in fermentor when fermented and cultured
DEG C, 0.03~0.05Mpa of pressure inside the tank, 5~6L/min of air mass flow, speed of agitator 500-700rpm.
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| CN101381747A (en) * | 2008-10-28 | 2009-03-11 | 中国人民解放军军事医学科学院野战输血研究所 | Method for extracting coenzyme Q10 from microorganism |
| CN102876743A (en) * | 2012-10-23 | 2013-01-16 | 上虞新和成生物化工有限公司 | Novel process for fermenting coenzyme Q10 based on online oxygen consumption rate control |
| CN103509816A (en) * | 2012-06-15 | 2014-01-15 | 浙江新和成股份有限公司 | Coenzyme-Q10-production engineered bacteria construction method, engineered bacteria, and application thereof |
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| CN101381747A (en) * | 2008-10-28 | 2009-03-11 | 中国人民解放军军事医学科学院野战输血研究所 | Method for extracting coenzyme Q10 from microorganism |
| CN103509816A (en) * | 2012-06-15 | 2014-01-15 | 浙江新和成股份有限公司 | Coenzyme-Q10-production engineered bacteria construction method, engineered bacteria, and application thereof |
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