CN109943599A - A kind of method of fermenting and producing long-chain biatomic acid - Google Patents
A kind of method of fermenting and producing long-chain biatomic acid Download PDFInfo
- Publication number
- CN109943599A CN109943599A CN201711379782.4A CN201711379782A CN109943599A CN 109943599 A CN109943599 A CN 109943599A CN 201711379782 A CN201711379782 A CN 201711379782A CN 109943599 A CN109943599 A CN 109943599A
- Authority
- CN
- China
- Prior art keywords
- fermentation
- acid
- fatty acids
- saturated fatty
- cctcc
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
- 239000002253 acid Substances 0.000 title claims abstract description 88
- 238000000034 method Methods 0.000 title claims abstract description 40
- 238000000855 fermentation Methods 0.000 claims abstract description 132
- 230000004151 fermentation Effects 0.000 claims abstract description 132
- 150000001335 aliphatic alkanes Chemical class 0.000 claims description 30
- 241001530515 Candida sake Species 0.000 claims description 26
- 238000009423 ventilation Methods 0.000 claims description 26
- 238000003756 stirring Methods 0.000 claims description 25
- 241000222178 Candida tropicalis Species 0.000 claims description 21
- 238000002156 mixing Methods 0.000 claims description 18
- 235000003441 saturated fatty acids Nutrition 0.000 claims description 17
- 239000000758 substrate Substances 0.000 claims description 17
- -1 saturated fatty acids ester Chemical class 0.000 claims description 12
- 150000003839 salts Chemical class 0.000 claims description 10
- 150000004671 saturated fatty acids Chemical class 0.000 claims description 10
- 125000005472 straight-chain saturated fatty acid group Chemical class 0.000 claims description 7
- DNXHEGUUPJUMQT-CBZIJGRNSA-N Estrone Chemical compound OC1=CC=C2[C@H]3CC[C@](C)(C(CC4)=O)[C@@H]4[C@@H]3CCC2=C1 DNXHEGUUPJUMQT-CBZIJGRNSA-N 0.000 claims 1
- 238000006243 chemical reaction Methods 0.000 abstract description 28
- 238000009776 industrial production Methods 0.000 abstract 1
- 238000004519 manufacturing process Methods 0.000 description 34
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 description 33
- 239000007788 liquid Substances 0.000 description 30
- 239000001963 growth medium Substances 0.000 description 28
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 24
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 24
- FGIUAXJPYTZDNR-UHFFFAOYSA-N potassium nitrate Chemical compound [K+].[O-][N+]([O-])=O FGIUAXJPYTZDNR-UHFFFAOYSA-N 0.000 description 24
- 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 description 19
- 239000008103 glucose Substances 0.000 description 19
- 238000011218 seed culture Methods 0.000 description 16
- 239000004215 Carbon black (E152) Substances 0.000 description 14
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 description 14
- 229930195733 hydrocarbon Natural products 0.000 description 14
- 229910052757 nitrogen Inorganic materials 0.000 description 12
- 235000010333 potassium nitrate Nutrition 0.000 description 12
- 239000004323 potassium nitrate Substances 0.000 description 12
- 229940041514 candida albicans extract Drugs 0.000 description 11
- 238000007254 oxidation reaction Methods 0.000 description 11
- 239000012138 yeast extract Substances 0.000 description 11
- 239000001888 Peptone Substances 0.000 description 10
- 108010080698 Peptones Proteins 0.000 description 10
- 150000002430 hydrocarbons Chemical class 0.000 description 10
- 229910000402 monopotassium phosphate Inorganic materials 0.000 description 10
- 235000019796 monopotassium phosphate Nutrition 0.000 description 10
- 235000019319 peptone Nutrition 0.000 description 10
- PJNZPQUBCPKICU-UHFFFAOYSA-N phosphoric acid;potassium Chemical compound [K].OP(O)(O)=O PJNZPQUBCPKICU-UHFFFAOYSA-N 0.000 description 10
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 9
- 229910052799 carbon Inorganic materials 0.000 description 9
- 239000002609 medium Substances 0.000 description 9
- 238000010899 nucleation Methods 0.000 description 9
- 238000011534 incubation Methods 0.000 description 8
- 230000003647 oxidation Effects 0.000 description 8
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 7
- OAICVXFJPJFONN-UHFFFAOYSA-N Phosphorus Chemical compound [P] OAICVXFJPJFONN-UHFFFAOYSA-N 0.000 description 7
- 239000003102 growth factor Substances 0.000 description 7
- 238000011081 inoculation Methods 0.000 description 7
- 229910052943 magnesium sulfate Inorganic materials 0.000 description 7
- 235000019341 magnesium sulphate Nutrition 0.000 description 7
- 229910052698 phosphorus Inorganic materials 0.000 description 7
- 239000011574 phosphorus Substances 0.000 description 7
- 230000007704 transition Effects 0.000 description 7
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 6
- 229910019142 PO4 Inorganic materials 0.000 description 6
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 6
- XSQUKJJJFZCRTK-UHFFFAOYSA-N Urea Chemical compound NC(N)=O XSQUKJJJFZCRTK-UHFFFAOYSA-N 0.000 description 6
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 description 6
- 239000010452 phosphate Substances 0.000 description 6
- QGZKDVFQNNGYKY-UHFFFAOYSA-O Ammonium Chemical compound [NH4+] QGZKDVFQNNGYKY-UHFFFAOYSA-O 0.000 description 5
- 229930006000 Sucrose Natural products 0.000 description 5
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 5
- NINIDFKCEFEMDL-UHFFFAOYSA-N Sulfur Chemical compound [S] NINIDFKCEFEMDL-UHFFFAOYSA-N 0.000 description 5
- 239000005864 Sulphur Substances 0.000 description 5
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 description 5
- 229910052921 ammonium sulfate Inorganic materials 0.000 description 5
- 235000011130 ammonium sulphate Nutrition 0.000 description 5
- 230000000694 effects Effects 0.000 description 5
- 239000000047 product Substances 0.000 description 5
- 239000005720 sucrose Substances 0.000 description 5
- YBJHBAHKTGYVGT-ZKWXMUAHSA-N (+)-Biotin Chemical compound N1C(=O)N[C@@H]2[C@H](CCCCC(=O)O)SC[C@@H]21 YBJHBAHKTGYVGT-ZKWXMUAHSA-N 0.000 description 4
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 4
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 4
- XEEYBQQBJWHFJM-UHFFFAOYSA-N Iron Chemical compound [Fe] XEEYBQQBJWHFJM-UHFFFAOYSA-N 0.000 description 4
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 description 4
- 125000001477 organic nitrogen group Chemical group 0.000 description 4
- LWIHDJKSTIGBAC-UHFFFAOYSA-K tripotassium phosphate Chemical compound [K+].[K+].[K+].[O-]P([O-])([O-])=O LWIHDJKSTIGBAC-UHFFFAOYSA-K 0.000 description 4
- SNRUBQQJIBEYMU-UHFFFAOYSA-N Dodecane Natural products CCCCCCCCCCCC SNRUBQQJIBEYMU-UHFFFAOYSA-N 0.000 description 3
- 239000004202 carbamide Substances 0.000 description 3
- 235000014113 dietary fatty acids Nutrition 0.000 description 3
- 235000019441 ethanol Nutrition 0.000 description 3
- 229930195729 fatty acid Natural products 0.000 description 3
- 239000000194 fatty acid Substances 0.000 description 3
- 150000004665 fatty acids Chemical class 0.000 description 3
- 229910052751 metal Inorganic materials 0.000 description 3
- 244000005700 microbiome Species 0.000 description 3
- 238000011160 research Methods 0.000 description 3
- OWEGMIWEEQEYGQ-UHFFFAOYSA-N 100676-05-9 Natural products OC1C(O)C(O)C(CO)OC1OCC1C(O)C(O)C(O)C(OC2C(OC(O)C(O)C2O)CO)O1 OWEGMIWEEQEYGQ-UHFFFAOYSA-N 0.000 description 2
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 description 2
- RYGMFSIKBFXOCR-UHFFFAOYSA-N Copper Chemical compound [Cu] RYGMFSIKBFXOCR-UHFFFAOYSA-N 0.000 description 2
- 108090000790 Enzymes Proteins 0.000 description 2
- 102000004190 Enzymes Human genes 0.000 description 2
- FYYHWMGAXLPEAU-UHFFFAOYSA-N Magnesium Chemical compound [Mg] FYYHWMGAXLPEAU-UHFFFAOYSA-N 0.000 description 2
- GUBGYTABKSRVRQ-PICCSMPSSA-N Maltose Natural products O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@@H](CO)OC(O)[C@H](O)[C@H]1O GUBGYTABKSRVRQ-PICCSMPSSA-N 0.000 description 2
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 description 2
- ZLMJMSJWJFRBEC-UHFFFAOYSA-N Potassium Chemical compound [K] ZLMJMSJWJFRBEC-UHFFFAOYSA-N 0.000 description 2
- QAOWNCQODCNURD-UHFFFAOYSA-L Sulfate Chemical compound [O-]S([O-])(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-L 0.000 description 2
- 240000008042 Zea mays Species 0.000 description 2
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 description 2
- 235000002017 Zea mays subsp mays Nutrition 0.000 description 2
- HCHKCACWOHOZIP-UHFFFAOYSA-N Zinc Chemical compound [Zn] HCHKCACWOHOZIP-UHFFFAOYSA-N 0.000 description 2
- 150000001413 amino acids Chemical class 0.000 description 2
- BIGPRXCJEDHCLP-UHFFFAOYSA-N ammonium bisulfate Chemical compound [NH4+].OS([O-])(=O)=O BIGPRXCJEDHCLP-UHFFFAOYSA-N 0.000 description 2
- ZRIUUUJAJJNDSS-UHFFFAOYSA-N ammonium phosphates Chemical compound [NH4+].[NH4+].[NH4+].[O-]P([O-])([O-])=O ZRIUUUJAJJNDSS-UHFFFAOYSA-N 0.000 description 2
- GUBGYTABKSRVRQ-QUYVBRFLSA-N beta-maltose Chemical compound OC[C@H]1O[C@H](O[C@H]2[C@H](O)[C@@H](O)[C@H](O)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@@H]1O GUBGYTABKSRVRQ-QUYVBRFLSA-N 0.000 description 2
- 238000011953 bioanalysis Methods 0.000 description 2
- 239000011616 biotin Substances 0.000 description 2
- 229960002685 biotin Drugs 0.000 description 2
- 235000020958 biotin Nutrition 0.000 description 2
- 239000011575 calcium Substances 0.000 description 2
- 229910052791 calcium Inorganic materials 0.000 description 2
- 235000013877 carbamide Nutrition 0.000 description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- 229910052802 copper Inorganic materials 0.000 description 2
- 239000010949 copper Substances 0.000 description 2
- 235000005822 corn Nutrition 0.000 description 2
- 238000006114 decarboxylation reaction Methods 0.000 description 2
- 239000003112 inhibitor Substances 0.000 description 2
- 239000013067 intermediate product Substances 0.000 description 2
- 229910052742 iron Inorganic materials 0.000 description 2
- 238000009630 liquid culture Methods 0.000 description 2
- 239000011777 magnesium Substances 0.000 description 2
- 229910052749 magnesium Inorganic materials 0.000 description 2
- WPBNNNQJVZRUHP-UHFFFAOYSA-L manganese(2+);methyl n-[[2-(methoxycarbonylcarbamothioylamino)phenyl]carbamothioyl]carbamate;n-[2-(sulfidocarbothioylamino)ethyl]carbamodithioate Chemical compound [Mn+2].[S-]C(=S)NCCNC([S-])=S.COC(=O)NC(=S)NC1=CC=CC=C1NC(=S)NC(=O)OC WPBNNNQJVZRUHP-UHFFFAOYSA-L 0.000 description 2
- 230000007246 mechanism Effects 0.000 description 2
- 235000013379 molasses Nutrition 0.000 description 2
- 229940094933 n-dodecane Drugs 0.000 description 2
- 239000011591 potassium Substances 0.000 description 2
- 229910052700 potassium Inorganic materials 0.000 description 2
- 229910000160 potassium phosphate Inorganic materials 0.000 description 2
- 235000011009 potassium phosphates Nutrition 0.000 description 2
- 239000000843 powder Substances 0.000 description 2
- 230000008569 process Effects 0.000 description 2
- YRWWOAFMPXPHEJ-OFBPEYICSA-K sodium L-ascorbic acid 2-phosphate Chemical compound [Na+].[Na+].[Na+].OC[C@H](O)[C@H]1OC(=O)C(OP([O-])([O-])=O)=C1[O-] YRWWOAFMPXPHEJ-OFBPEYICSA-K 0.000 description 2
- 229940048058 sodium ascorbyl phosphate Drugs 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 230000009466 transformation Effects 0.000 description 2
- 239000011782 vitamin Substances 0.000 description 2
- 229940088594 vitamin Drugs 0.000 description 2
- 229930003231 vitamin Natural products 0.000 description 2
- 235000013343 vitamin Nutrition 0.000 description 2
- 150000003722 vitamin derivatives Chemical class 0.000 description 2
- 239000011701 zinc Substances 0.000 description 2
- 229910052725 zinc Inorganic materials 0.000 description 2
- AKUVRZKNLXYTJX-UHFFFAOYSA-N 3-benzylazetidine Chemical compound C=1C=CC=CC=1CC1CNC1 AKUVRZKNLXYTJX-UHFFFAOYSA-N 0.000 description 1
- 241000894006 Bacteria Species 0.000 description 1
- 241000222120 Candida <Saccharomycetales> Species 0.000 description 1
- 229920002261 Corn starch Polymers 0.000 description 1
- 239000004831 Hot glue Substances 0.000 description 1
- OFOBLEOULBTSOW-UHFFFAOYSA-N Malonic acid Chemical compound OC(=O)CC(O)=O OFOBLEOULBTSOW-UHFFFAOYSA-N 0.000 description 1
- 229910002651 NO3 Inorganic materials 0.000 description 1
- NHNBFGGVMKEFGY-UHFFFAOYSA-N Nitrate Chemical compound [O-][N+]([O-])=O NHNBFGGVMKEFGY-UHFFFAOYSA-N 0.000 description 1
- GRYLNZFGIOXLOG-UHFFFAOYSA-N Nitric acid Chemical compound O[N+]([O-])=O GRYLNZFGIOXLOG-UHFFFAOYSA-N 0.000 description 1
- 239000004952 Polyamide Substances 0.000 description 1
- 229910000831 Steel Inorganic materials 0.000 description 1
- XJLXINKUBYWONI-DQQFMEOOSA-N [[(2r,3r,4r,5r)-5-(6-aminopurin-9-yl)-3-hydroxy-4-phosphonooxyoxolan-2-yl]methoxy-hydroxyphosphoryl] [(2s,3r,4s,5s)-5-(3-carbamoylpyridin-1-ium-1-yl)-3,4-dihydroxyoxolan-2-yl]methyl phosphate Chemical compound NC(=O)C1=CC=C[N+]([C@@H]2[C@H]([C@@H](O)[C@H](COP([O-])(=O)OP(O)(=O)OC[C@@H]3[C@H]([C@@H](OP(O)(O)=O)[C@@H](O3)N3C4=NC=NC(N)=C4N=C3)O)O2)O)=C1 XJLXINKUBYWONI-DQQFMEOOSA-N 0.000 description 1
- 238000009825 accumulation Methods 0.000 description 1
- 150000007513 acids Chemical class 0.000 description 1
- 125000001931 aliphatic group Chemical group 0.000 description 1
- DCAYPVUWAIABOU-UHFFFAOYSA-N alpha-n-hexadecene Natural products CCCCCCCCCCCCCCCC DCAYPVUWAIABOU-UHFFFAOYSA-N 0.000 description 1
- 229910000147 aluminium phosphate Inorganic materials 0.000 description 1
- 239000013556 antirust agent Substances 0.000 description 1
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 1
- 230000001580 bacterial effect Effects 0.000 description 1
- 230000000903 blocking effect Effects 0.000 description 1
- 239000006227 byproduct Substances 0.000 description 1
- BVKZGUZCCUSVTD-UHFFFAOYSA-N carbonic acid Chemical compound OC(O)=O BVKZGUZCCUSVTD-UHFFFAOYSA-N 0.000 description 1
- 239000003054 catalyst Substances 0.000 description 1
- 239000003638 chemical reducing agent Substances 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 229960001657 chlorpromazine hydrochloride Drugs 0.000 description 1
- 239000011248 coating agent Substances 0.000 description 1
- 238000000576 coating method Methods 0.000 description 1
- 239000005515 coenzyme Substances 0.000 description 1
- 239000008120 corn starch Substances 0.000 description 1
- 230000007423 decrease Effects 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 230000001419 dependent effect Effects 0.000 description 1
- 150000001991 dicarboxylic acids Chemical class 0.000 description 1
- 238000004090 dissolution Methods 0.000 description 1
- 230000002708 enhancing effect Effects 0.000 description 1
- 238000001704 evaporation Methods 0.000 description 1
- 230000008020 evaporation Effects 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 239000003205 fragrance Substances 0.000 description 1
- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 description 1
- 229910052737 gold Inorganic materials 0.000 description 1
- 239000010931 gold Substances 0.000 description 1
- 230000006698 induction Effects 0.000 description 1
- JEIPFZHSYJVQDO-UHFFFAOYSA-N iron(III) oxide Inorganic materials O=[Fe]O[Fe]=O JEIPFZHSYJVQDO-UHFFFAOYSA-N 0.000 description 1
- IIYFAKIEWZDVMP-UHFFFAOYSA-N linear paraffin C13 Natural products CCCCCCCCCCCCC IIYFAKIEWZDVMP-UHFFFAOYSA-N 0.000 description 1
- 239000000314 lubricant Substances 0.000 description 1
- 230000000813 microbial effect Effects 0.000 description 1
- 230000002906 microbiologic effect Effects 0.000 description 1
- 238000002703 mutagenesis Methods 0.000 description 1
- 231100000350 mutagenesis Toxicity 0.000 description 1
- 230000035772 mutation Effects 0.000 description 1
- 229910017604 nitric acid Inorganic materials 0.000 description 1
- 238000010525 oxidative degradation reaction Methods 0.000 description 1
- 229910052760 oxygen Inorganic materials 0.000 description 1
- 239000001301 oxygen Substances 0.000 description 1
- 239000003973 paint Substances 0.000 description 1
- 239000004014 plasticizer Substances 0.000 description 1
- 229920002647 polyamide Polymers 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 230000005562 seed maturation Effects 0.000 description 1
- 238000007086 side reaction Methods 0.000 description 1
- 239000010959 steel Substances 0.000 description 1
- 238000005728 strengthening Methods 0.000 description 1
- 230000009897 systematic effect Effects 0.000 description 1
- 238000010998 test method Methods 0.000 description 1
- 230000003519 ventilatory effect Effects 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
- 230000003313 weakening effect Effects 0.000 description 1
Landscapes
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention discloses a kind of method of fermenting and producing long-chain biatomic acid, this method are as follows: during the fermentation, control rate of producing acid is 1.0~2.0g/L/h.Method of the invention can significantly improve the fermentation conversion rate of long-chain biatomic acid, and low in cost, easy to operate, be suitble to large-scale industrial production.
Description
Technical field
The present invention relates to fermentation arts, and in particular to a kind of method of fermenting and producing long-chain biatomic acid.
Background technique
Long-chain biatomic acid has very extensive purposes, can synthesize extraordinary polyamide, height by raw material of long-chain biatomic acid
Grade fragrance, high-grade hot melt adhesive, cold resistant plasticizer, senior lubricant, advanced antirust agent, advanced paint and coating etc..Long-chain binary
Acid can usually be synthesized with chemical method or bioanalysis.Chemical method synthetic route is long, and reaction needs high temperature and pressure, wants to catalyst
Ask comparison harsh, therefore long-chain biatomic acid at industrial scale is of less types, only a small number of product such as 12 carbon long-chain biatomic acids
Kind.And bioanalysis is to be obtained using long chain alkane as substrate by microorganism conversion, utilizes the special function of microorganism diterminal oxidation
Long-chain biatomic acid can be prepared by fermenting substrate of n-alkane.The mechanism study of long-chain biatomic acid is generated by microbiological oxidation alkane
Known to: its main reaction is α, and omega oxidation, side reaction is alpha-oxidation decarboxylation and beta oxidation, makes the product long-chain biatomic acid quilt generated
Further oxidative degradation.Beta oxidation, which is mainly grown to microorganism, provides energy, maintains the conversion activity of microbial cell, this just leads
Cause substrate alkane that cannot be completely converted into product binary acid, and the conversion ratio of alkane is to Guan Chong for the production cost of binary acid
It wants.
Chinese patent CN102808004A and CN1259424C disclose a kind of side for improving fermentation yield of long-chain dicarboxylic acid
Method.This method improves fermentation conversion rate by adding alpha-oxidation decarboxylation or beta oxidation inhibitor during the fermentation, uses
Inhibitor is the reagents such as chlorpromazine hydrochloride, halogenated aliphatic acid, and such reagent price is expensive, increases the extraction and purification process of binary acid
And cost, and conversion ratio highest is only increased to 89% (w/w).Chinese patent CN105755062A discloses a kind of utilize and aoxidizes also
The method that former current potential regulates and controls fermentation process production long-chain biatomic acid.This method combines oxygen by adjusting ventilatory capacity, speed of agitator
Agent and reducing agent realize that two stages ORP of long-chain biatomic acid control fermentation, but do not refer to binary acid yield.Chinese patent
CN103074325A discloses a kind of method of mutagenesis of the Candida tropicalis of long-chain biatomic acid, and this method utilizes induction mutation of bacterium
Superior strain is screened, but does not refer to binary acid conversion ratio.The document of foreign study long-chain biatomic acid is to carry out base to bacterial strain mostly
Because of transformation, by blocking or weakening the related enzyme systems of fatty acid beta oxidation, the enzyme system for strengthening fatty acid α-ω oxidation produces to improve
The yield of product.
In conclusion the technique of Production of Long-chain Dicarboxylic Acids by Fermentation Methods in the prior art, fermentation conversion rate is not generally high, and each
Kind improves the method higher cost of long-chain biatomic acid fermentation conversion rate, and effect is not very significant, does not have still in the prior art
One kind can efficiently improve long-chain biatomic acid fermentation conversion rate, and method easy to operate, at low cost.
Summary of the invention
It is not high in order to solve long-chain biatomic acid fermentation conversion rate existing in the prior art, in order to improve fermentation conversion rate,
It needs to increase great amount of cost and effect and inapparent defect, a kind of method of fermenting and producing long-chain biatomic acid is provided, the present invention
Method can significantly improve the fermentation conversion rate of long-chain biatomic acid, and it is low in cost, it is easy to operate, be suitble to heavy industrialization
Production.
The purpose of the present invention is to provide a kind of method of fermenting and producing long-chain biatomic acid, in the fermentation process, acid is produced
Rate is 1.0~2.0g/L/h, preferably 1.1~1.7g/L/h, more preferable 1.2~1.5g/L/h.
Those skilled in the art understand, and the rate of producing acid refers to the throughput rate of long-chain biatomic acid in fermentation process,
The fermentation and acid speed is characterized with long-chain biatomic acid quality that unit time unit volume fermented feed liquid generates.
In the prior art, there are many researchs to the method for improving long-chain biatomic acid fermentation conversion rate, but there has been no pass through control
Fermentation process acid production speed processed just improves the report of fermentation conversion rate.Inventor passes through to all of long-chain biatomic acid zymotechnique
More researchs, discovery: acid production speed has a great impact to the fermentation conversion rate of long-chain biatomic acid.By the way that acid production speed is controlled
In a specific range, it can effectively improve the fermentation conversion rate of long-chain biatomic acid；On this basis, inventor is in terms of mechanism
The deep structure research phenomenon, discovery: why acid production speed have a significant impact fermentation conversion rate, is because of cell density pair
Conversion, intermediate product, generation of by-product of substrate etc. have an impact, and only under certain equilibrium condition, are just able to achieve high
The fermentation conversion rate of long-chain biatomic acid；One side acid production speed is too fast, and substrate conversion rate is too fast, and coenzyme NADP 11 is insufficient,
Beta-oxidation enhancing, causes the substrate transformation rate low, and the accumulation of intermediate product hydroxycarboxylic acid excessively causes product quality to decline；Another party
Face acid production speed is excessively slow, and fermentation period extension causes to produce strength reduction, increases production cost.Therefore the reasonable acid production speed of control
The fermentation conversion rate of long-chain biatomic acid can be effectively improved.
More than, the preferred technical solution of scheme, is described further in view of the above technology.
The present invention to the control method of acid production speed without limitation, it is only necessary to control acid production speed in particular range i.e.
It can；In general, the method for conventional control acid production speed, such as: control stirring operation, control fermentation temperature or control are logical
Air quantity or the combination of several control means etc. are suitable for the present invention, cooperate other technological operations, common to coordinate, Neng Goushi
Existing technical effect of the invention.
A preferred technical solution of the invention can cooperate other process conditions, by controlling mixing speed to control
Acid production speed processed.Those skilled in the art understand, the determination of mixing speed dependent on fermentor pot, the shape of agitating paddle,
The factors such as quantity.
A preferred technical solution of the invention, the speed of agitator when fermentation are 500~1000rpm.It is described to stir
It mixes speed to characterize with the speed of agitator of two layers of turbine disc blade stirring paddle, more specifically, the mixing speed is stainless with 10L
It is characterized on steel fermentor using the speed of agitator of two layers of turbine disc blade stirring paddle.
Herein it should be noted that the present invention is not limited to use above-mentioned agitating paddle, it is also not necessarily limited to above-mentioned mixing speed, as long as
The agitating paddle of this field routine cooperates specific speed, can be realized specific acid production speed, in protection scope of the present invention
It is interior.
The preferred another technical solution of of the invention one can also cooperate other process conditions, come by controlling ventilation quantity
Control acid production speed.
A preferred technical solution of the invention, the ventilation quantity in the fermentation process are 0.1~1vvm, preferably 0.2
~0.6vvm；The ventilation quantity is characterized with the volume of air (L) that is passed through in unit time (min) unit volume (L) fermentation liquid.
The present invention not only can control acid production speed through the above way, can also be come by the combination of several ways
Control acid production speed.
A preferred technical solution of the invention is matched by controlling the common adjusting of the mixing speed and ventilation quantity
Other fermentation parameters are closed, can be realized the effect for preferably improving fermentation conversion rate.The mixing speed and ventilation quantity are full
Sufficient the following conditions:
0.6-0.00042A≤F≤0.8-0.00038A；
Wherein: A indicates that the numerical value of mixing speed, the numerical value are the numerical value as unit of rpm；The mixing speed is with two layers
The speed of agitator of turbine disc blade stirring paddle characterizes；
F indicates that the numerical value of ventilation quantity, the numerical value are the numerical value as unit of vvm.
A preferred technical solution of the invention, the strain of the fermentation include candida tropicalis (Candida
) or candida sake (Candidasake) Tropicalis.Such as: it can be candida tropicalis (Candida
Tropicalis) (deposit number is CCTCC M203052) or candida tropicalis (Candida Tropicalis)
CATN145 (deposit number is CCTCC M 2011192) or candida sake (Candidasake) CATH4013 (deposit number
For CCTCC M2011486) or candida sake (Candidasake) CATH4014 (deposit number CCTCC
M2011487) perhaps candida sake (Candidasake) CATH4012 (deposit number be CCTCC M2011485) or
Candida sake (Candidasake) CATH4016 (deposit number is CCTCC M2011488) or candida sake
(Candidasake) CATH430 (deposit number is CCTCC M2011489).
A preferred technical solution of the invention, the substrate of the fermentation include the normal alkane of C9~C18, linear saturation
One of fatty acid, linear saturated fatty acids ester and salts of straight-chain saturated fatty acids are a variety of；Preferably include C11~C16 just
One of alkane, linear saturated fatty acids, linear saturated fatty acids ester and salts of straight-chain saturated fatty acids are a variety of；More preferably
Normal alkane, linear saturated fatty acids, linear saturated fatty acids ester and the linear saturation rouge of C11, C12, C13, C15, C14 or C16
One of fat hydrochlorate.
A preferred technical solution of the invention, the culture medium of the fermentation include: carbon source, nitrogen source, phosphorus source, micro gold
Belong to element source and growth factor.
Wherein, the carbon source includes either one of glucose, sucrose, maltose, molasses, methanol, ethyl alcohol or more
Kind；It is highly preferred that the carbon source include either glucose, sucrose it is one or more.The carbon source concentration preferably 20~
60g/L。
Wherein, the nitrogen source includes either organic nitrogen and/or inorganic nitrogen, and organic nitrogen includes but is not limited to yeast extract, egg
One of white peptone, corn pulp are a variety of, and inorganic nitrogen includes but is not limited to one of urea, ammonium sulfate, potassium nitrate or a variety of；
Preferably, the nitrogen source includes either ammonium sulfate and/or potassium nitrate.The concentration of the nitrogen source preferably 0.5~10g/L.It is preferred that
Ground, the concentration of the ammonium sulfate preferably 0.5~5g/L.Preferably, the concentration of the potassium nitrate preferably 0.5~5g/L.
Wherein, phosphorus source includes either one of phosphate, dibasic alkaliine and dihydric phosphate or a variety of；It is excellent
Selection of land, the phosphate include either one of potassium phosphate, sodium ascorbyl phosphate, ammonium phosphate salt or a variety of；It is highly preferred that institute
It states phosphate and includes either potassium dihydrogen phosphate.The concentration of phosphorus source preferably 1~5g/L.
Wherein, the minor metallic element source includes potassium, calcium, magnesium, iron, copper, zinc, the sulfate of manganese, hydrochloride and nitric acid
One of salt is a variety of.The concentration preferably 0.1~50ppm in the minor metallic element source.
Wherein, the growth factor includes one of amino acid, citric acid and vitamin or a variety of；It is highly preferred that institute
State the mixing that growth factor includes either one or both of citric acid, biotin.The growth factor concentration is preferred
0.01~1ppm.
A preferred technical solution of the invention, the tank temperature when fermentation are 28~32 DEG C.
A preferred technical solution of the invention, the tank pressure when fermentation is 0.05~0.14Mpa, and the pressure is
Gauge pressure.
A preferred technical solution of the invention, the ferment control pH are 5.0~8.5.
Those skilled in the art are according to common sense in the field, it can be determined that the stage of fermentation.In general, before the fermentation
Phase is that fermentation starts to 30~60h of fermentation.The fermentation later period is that fermentation starts 30~60h to fermentation ends.
A preferred technical solution of the invention, the fermentation period are 100~180h.
Specific embodiment
The present invention provides a kind of side that long-chain biatomic acid fermentation conversion rate is improved by control fermentation process rate of producing acid
Method.Wherein, the rate of producing acid refers to the throughput rate of long-chain biatomic acid in fermentation process, and the fermentation and acid speed is with list
The long-chain biatomic acid quality that position chronomere volume fermented feed liquid generates characterizes.
.The present invention by control fermentation process ventilation quantity and/or fermentation process mixing speed, cooperate other technological parameters and
Condition, to control fermentation process rate of producing acid in a suitable range, to improve fermentation conversion rate.
The method of fermenting and producing long-chain biatomic acid, comprising the following steps:
A) actication of culture；
B seed liquor) is prepared in seeding tank using seed culture medium；
C) seed liquor is inoculated into the fermentor containing fermentation medium, in fermentation process, rate of producing acid be 1.0~
2.0g/L/h, preferably 1.1~1.7g/L/h, more preferable 1.2~1.5g/L/h；Rate of producing acid refers to long-chain biatomic acid in fermentation process
Throughput rate, by unit time unit volume fermented feed liquid generate long-chain biatomic acid quality in terms of.
In one embodiment of the present of invention, A) actication of culture is the following steps are included: take candida tropicalis (Candida
Tropicalis) or the glycerol stock of candida sake (Candidasake) is in the shaking flask of YPD culture medium, shaking table culture, turns
200~250rpm of speed, cultivates 24~48h by 28~30 DEG C of cultivation temperature.
Step A) in, YPD culture medium includes: peptone 10g/kg, yeast extract 5g/kg, and glucose 10g/kg, pH are natural.
In one embodiment of the present of invention, step B) in the parameter of seeding tank be: 28~30 DEG C of cultivation temperature, ventilation quantity
0.1~1vvm, preferably 0.2~0.6vvm；Tank pressure is 0.08~0.1MPa (gauge pressure), and 12~36h of incubation time, seed maturation refers to
It is designated as OD620It is 15~30.
Step B) in seed culture medium be water-soluble liquid culture medium, preferably include following component: 10~30g/L of sucrose, corn
Starch 1.5~10g/L, 1~10g/L of yeast extract, 4~12g/L of potassium dihydrogen phosphate, 0.5~5g/L of urea, 0~30ml/ of alkane
L, water-soluble liquid culture medium is prepared with water, and is sterilized.
In one embodiment of the present of invention, step C) in, 28~32 DEG C of fermentation processes temperature, ventilation quantity 0.1~
1vvm, preferably 0.2~0.6vvm (volume of air that unit time unit volume fermentation liquid is passed through), tank pressure for 0.05~
0.14MPa (gauge pressure), with two layers of turbine disc blade stirring paddle stirring, 500~1000rpm of mixing speed；Add 10%~
The pH value 5.0~8.5 of the NaOH solution control fermentation liquid of 40% (w/v)；Start to add in batches when fermentation period is 10~20h
Enter substrate, total fermentation period is 100~180h；
In one embodiment of the present of invention, step C) in, substrate include the normal alkane of C9~C18, linear saturated fatty acids,
One of linear saturated fatty acids ester and salts of straight-chain saturated fatty acids are a variety of；Preferably include the normal alkane, straight of C11~C16
One of chain saturated fatty acid, linear saturated fatty acids ester and salts of straight-chain saturated fatty acids are a variety of；More preferably C11,
Normal alkane, linear saturated fatty acids, linear saturated fatty acids ester and the linear saturated fatty acids of C12, C13, C15, C14 or C16
One of salt；
In one embodiment of the present of invention, step C) in fermentation medium, comprising: carbon source, nitrogen source, phosphorus source, trace meter
Element source, growth factor；
Carbon source includes either one of glucose, sucrose, maltose, molasses, methanol, ethyl alcohol or a variety of；More preferably
Ground, carbon source include one or two kinds of mixing of either glucose, sucrose；Carbon source concentration is 20~60g/L.
Nitrogen source can be organic nitrogen and/or inorganic nitrogen, and organic nitrogen includes but is not limited to yeast extract, peptone, in corn pulp
It is one or more, inorganic nitrogen includes but is not limited to one of urea, ammonium sulfate, potassium nitrate or a variety of；Preferably, nitrogen source packet
Include one or two kinds of mixing of either ammonium sulfate, potassium nitrate；The concentration of nitrogen source is 0.5~5g/L.
Phosphorus source includes either one of phosphoric acid normal salt, dibasic alkaliine and dihydric phosphate or a variety of, preferably phosphorus
Hydrochlorate includes either one of potassium phosphate, sodium ascorbyl phosphate, ammonium phosphate salt or a variety of；It is highly preferred that phosphate include or
Person is potassium dihydrogen phosphate.Phosphorus source concentration is 1~5g/L.
Minor metallic element source includes one of potassium, calcium, magnesium, iron, copper, zinc, the sulfate of manganese, hydrochloride and nitrate
Or it is a variety of.The concentration in microelement source is 0.1~50ppm.
Growth factor includes one of amino acid, citric acid and vitamin or a variety of；Preferably include either citric acid,
The mixing of one or both of biotin.Growth factor concentration is 0.01~1ppm.
Below by embodiment, the present invention is described in detail, so that the features and advantages of the present invention become apparent from.But it answers
This points out that for embodiment for understanding design of the invention, the scope of the present invention is not limited only to reality listed herein
Apply example.
It is such as not particularly illustrated, concentration of the present invention is mass percent concentration.
In the present invention, the binary acid concentration in culture solution is measured, technology well known to those skilled in the art, example can be used
The measuring method as disclosed in Chinese patent ZL 95117436.3.Specifically, the pH to 3.0 of fermentation liquid is adjusted with hydrochloric acid solution,
Then plus 100mL ether is for the binary acid in extractive fermentation liquid, then uses evaporation to remove ether, obtains binary acid powder；
In ethanol by the dissolution of obtained binary acid powder, and with the NaOH solution of 0.1mol/L it titrates, finally obtains in fermentation liquid
Binary acid titer.
In the present invention, unless otherwise indicated, " about " Lai Xiuzheng is used in the parameters such as component, reaction condition, concentration
Numerical value.Therefore numerical parameter in the specification and in the claims is an approximation, depends on the desired property of the present invention
Energy.At least, it is not considered as the limitation of doctrine of equivalents application protected to the claims in the present invention.At least, the number of each parameter
With rounding up, the digit of effective digital historically determines value.Although numerical value in a particular embodiment is as far as possible
It is accurate to accomplish, but since the systematic error of experimental test procedures necessarily causes any data that can all have certain error.
Embodiment 1
(1) actication of culture:
The glycerol tube strain of candida tropicalis is taken to be inoculated in the shaking flask equipped with 30ml seed culture medium, pH is naturally, 29
At DEG C, 220rpm shaking table culture is for 24 hours；YPD culture medium includes: peptone 10g/kg, yeast extract 5g/kg, glucose 10g/kg；
(2) seed liquor is prepared:
Take shake-flask seed access equipped in the seeding tank of seed culture medium, the initial ph value of system is 6.0,29 DEG C after inoculation
Under, ventilation quantity 0.5vvm, tank presses 0.1MPa, cultivates 18h, and pH drops to 3, OD naturally in incubation620It grows to 15；
(3) it ferments:
Seed is inoculated into containing 6L fermentation medium (glucose 50g/L, potassium nitrate 3g/L, potassium dihydrogen phosphate 4g/L, sulphur
Sour ammonium 0.3g/L, magnesium sulfate 50ppm, citric acid 1ppm) fermentor in, 30 DEG C of fermentation processes temperature, ventilation quantity
0.5vvm, tank pressure is about 0.1MPa (gauge pressure), with two layers of turbine disc blade stirring paddle stirring, speed of agitator 1000rpm；Fermentation
Process adds the pH value of the liquid alkaline control fermentation liquid of 30% (w/v)；When fermentation period is 10~20h start that substrate is added portionwise
N-dodecane hydrocarbon, control fermentation liquid in Determination of Alkane Content 10% (v/v) below；Ferment control pH 5.0-8.5, total fermentation period are
130 hours, whens fermentation ends, residual hydrocarbon content was essentially 0；
The production concentration 130mg/g of LCDA, acid production speed 1.9g/L/h, alkane mass transitions rate 85%.
Embodiment 2
(1) actication of culture:
The glycerol tube strain of candida tropicalis is taken to be inoculated in the shaking flask equipped with 30ml seed culture medium, pH is naturally, 29
At DEG C, 220rpm shaking table culture is for 24 hours；YPD culture medium includes: peptone 10g/kg, yeast extract 5g/kg, glucose 10g/kg；
(2) seed liquor is prepared:
Take shake-flask seed access equipped in the seeding tank of seed culture medium, the initial ph value of system is 6.0,29 DEG C after inoculation
Under, ventilation quantity 0.5vvm, tank presses 0.1MPa, cultivates 18h, and pH drops to 3, OD naturally in incubation620It grows to 15；
(3) it ferments:
Seed is inoculated into containing fermentation medium (glucose 20g/L, potassium nitrate 2g/L, potassium dihydrogen phosphate 3g/L, sulfuric acid
Ammonium 2g/L, magnesium sulfate 50ppm, citric acid 1ppm) fermentor in, 30 DEG C of fermentation processes temperature, ventilation quantity 1.0vvm, tank
Pressure is about 0.12MPa (gauge pressure), and with two layers of turbine disc blade stirring paddle stirring, speed of agitator 600rpm, fermentation process is added
The pH value 5.0-8.5 of the liquid alkaline control fermentation liquid of 30% (w/v)；Start point 6 batches of addition substrates when fermentation period is 10~20h
N-dodecane hydrocarbon, control fermentation liquid in Determination of Alkane Content 10% (v/v) below；Total fermentation period is 140 hours, and whens fermentation ends is residual
Hydrocarbon content is essentially 0；
The production acid concentration 132mg/g of LCDA, acid production speed 1.8g/L/h, alkane mass transitions rate 88%.
Embodiment 3
(1) actication of culture:
The glycerol tube strain of candida tropicalis is taken to be inoculated in the shaking flask equipped with 30ml seed culture medium, pH is naturally, 29
At DEG C, 220rpm shaking table culture is for 24 hours；YPD culture medium includes: peptone 10g/kg, yeast extract 5g/kg, glucose 10g/kg；
(2) seed liquor is prepared:
Take shake-flask seed access equipped in the seeding tank of seed culture medium, the initial ph value of system is 6.0,29 DEG C after inoculation
Under, ventilation quantity 0.5vvm, tank presses 0.1MPa, cultivates 18h, and pH drops to 3, OD naturally in incubation620It grows to 15；
(3) it ferments:
Seed is inoculated into containing fermentation medium (glucose 30g/L, potassium nitrate 2g/L, potassium dihydrogen phosphate 3g/L, sulfuric acid
Ammonium 2g/L, magnesium sulfate 50ppm, citric acid 1ppm) fermentor in, 30 DEG C of fermentation processes temperature, ventilation quantity 0.2vvm, tank
Pressure is about 0.12MPa (gauge pressure), with two layers of turbine disc blade stirring paddle stirring, speed of agitator 1000rpm；Fermentation process is added
The pH value 5.0-8.5 of the liquid alkaline control fermentation liquid of 30% (w/v)；When fermentation period is 10~20h start that substrate is added portionwise
N-dodecane hydrocarbon, control fermentation liquid in Determination of Alkane Content 10% (v/v) below；Total fermentation period is 145 hours, and whens fermentation ends is residual
Hydrocarbon content is essentially 0；
The production acid concentration 150mg/g of LCDA, acid production speed 1.45g/L/h, alkane mass transitions rate 92%.
Embodiment 4
(1) actication of culture:
The glycerol tube strain of candida tropicalis is taken to be inoculated in the shaking flask equipped with 30ml seed culture medium, pH is naturally, 29
At DEG C, 220rpm shaking table culture is for 24 hours；YPD culture medium includes: peptone 10g/kg, yeast extract 5g/kg, glucose 10g/kg；
(2) seed liquor is prepared:
Take shake-flask seed access equipped in the seeding tank of seed culture medium, the initial ph value of system is 6.0,29 DEG C after inoculation
Under, ventilation quantity 0.5vvm, tank presses 0.1MPa, cultivates 18h, and pH drops to 3, OD naturally in incubation620It grows to 15；
(3) it ferments:
Seed is inoculated into containing fermentation medium (glucose 30g/L, potassium nitrate 2g/L, potassium dihydrogen phosphate 2.5g/L, sulphur
Sour ammonium 1.5g/L, magnesium sulfate 50ppm, citric acid 1ppm) fermentor in, 30 DEG C of fermentation processes temperature, ventilation quantity
0.4vvm, tank pressure is about 0.12MPa (gauge pressure), with two layers of turbine disc blade stirring paddle stirring, speed of agitator 600rpm；Fermentation
Process adds the pH value 5.0-8.5 of the liquid alkaline control fermentation liquid of 30% (w/v), starts in batches when fermentation period is 10~20h
Be added substrate n-dodecane hydrocarbon, control fermentation liquid in Determination of Alkane Content 10% (v/v) below；Total fermentation period is 160 hours, fermentation
At the end of residual hydrocarbon content be essentially 0；
Production the acid concentration 160mg/g, acid production speed 1.3g/L/h of LCDA.Alkane mass transitions rate 94%.
Embodiment 5
(1) actication of culture:
The glycerol tube strain of candida tropicalis is taken to be inoculated in the shaking flask equipped with 30ml seed culture medium, pH is naturally, 29
At DEG C, 220rpm shaking table culture is for 24 hours；YPD culture medium includes: peptone 10g/kg, yeast extract 5g/kg, glucose 10g/kg；
(2) seed liquor is prepared:
Take shake-flask seed access equipped in the seeding tank of seed culture medium, the initial ph value of system is 6.0,29 DEG C after inoculation
Under, ventilation quantity 0.5vvm, tank presses 0.1MPa, cultivates 18h, and pH drops to 3, OD naturally in incubation620It grows to 15；
(3) it ferments:
Seed is inoculated into containing fermentation medium (glucose 20g/L, potassium nitrate 2g/L, potassium dihydrogen phosphate 2.5g/L, sulphur
Sour ammonium 1.5g/L, magnesium sulfate 50ppm, citric acid 1ppm) fermentor in, 30 DEG C of fermentation processes temperature, ventilation quantity
0.3vvm, tank pressure is about 0.12MPa (gauge pressure), with two layers of turbine disc blade stirring paddle stirring, speed of agitator 900rpm；Fermentation
Process adds the pH value 5.0-8.5 of the liquid alkaline control fermentation liquid of 30% (w/v), starts points 7 batches when fermentation period is 10~20h
Be added substrate n-dodecane hydrocarbon, control fermentation liquid in Determination of Alkane Content 10% (v/v) below；Total fermentation period is 160 hours, fermentation
At the end of residual hydrocarbon content be essentially 0；
The production acid concentration 160mg/g of LCDA, acid production speed 1.4g/L/h, alkane mass transitions rate 95%.
Embodiment 6
(1) actication of culture:
The glycerol tube strain of candida tropicalis is taken to be inoculated in the shaking flask equipped with 30ml seed culture medium, pH is naturally, 29
At DEG C, 220rpm shaking table culture is for 24 hours；YPD culture medium includes: peptone 10g/kg, yeast extract 5g/kg, glucose 10g/kg；
(2) seed liquor is prepared:
Take shake-flask seed access equipped in the seeding tank of seed culture medium, the initial ph value of system is 6.0,29 DEG C after inoculation
Under, ventilation quantity 0.5vvm, tank presses 0.1MPa, cultivates 18h, and pH drops to 3, OD naturally in incubation620It grows to 15；
(3) it ferments:
Seed is inoculated into containing fermentation medium (glucose 20g/L, potassium nitrate 2g/L, potassium dihydrogen phosphate 2.5g/L, sulphur
Sour ammonium 1.5g/L, magnesium sulfate 50ppm, citric acid 1ppm) fermentor in, be added 10% (v/v) alkane, fermentation processes
30 DEG C of temperature, ventilation quantity 0.3vvm, tank pressure is about 0.12MPa (gauge pressure), with two layers of turbine disc blade stirring paddle stirring, stirring
Revolving speed 800rpm；Fermentation process adds the pH value 5.0-8.5 of the liquid alkaline control fermentation liquid of 30% (w/v), is 10 in fermentation period
Start that substrate n-tridecane hydrocarbon is added portionwise when~20h, controls Determination of Alkane Content in fermentation liquid and be no more than 10% (v/v) below；Always
Fermentation period is 160 hours, and whens fermentation ends, residual hydrocarbon content was essentially 0；
Production the acid concentration 150mg/g, acid production speed 1.25g/L/h of LCDA；Alkane mass transitions rate 93%.
Embodiment 7
(1) actication of culture:
The glycerol tube strain of candida tropicalis is taken to be inoculated in the shaking flask equipped with 30ml seed culture medium, pH is naturally, 29
At DEG C, 220rpm shaking table culture is for 24 hours；YPD culture medium includes: peptone 10g/kg, yeast extract 5g/kg, glucose 10g/kg；
(2) seed liquor is prepared:
Take shake-flask seed access equipped in the seeding tank of seed culture medium, the initial ph value of system is 6.0,29 DEG C after inoculation
Under, ventilation quantity 0.5vvm, tank presses 0.1MPa, cultivates 18h, and pH drops to 3, OD naturally in incubation620It grows to 15；
(3) it ferments:
Seed is inoculated into containing fermentation medium (glucose 20g/L, potassium nitrate 2g/L, potassium dihydrogen phosphate 2.5g/L, sulphur
Sour ammonium 1.5g/L, magnesium sulfate 50ppm, citric acid 1ppm) fermentor in, be added 10% (v/v) alkane, fermentation processes
30 DEG C of temperature, ventilation quantity 0.3vvm, tank pressure is about 0.12MPa (gauge pressure), with two layers of turbine disc blade stirring paddle stirring, stirring
Revolving speed 800rpm；Fermentation process adds the pH value 5.0-8.5 of the liquid alkaline control fermentation liquid of 30% (w/v), is 10 in fermentation period
Start that substrate hexadecane hydrocarbon is added portionwise when~20h, controls Determination of Alkane Content in fermentation liquid and be no more than 10% (v/v) below；Always
Fermentation period is 160 hours, and whens fermentation ends, residual hydrocarbon content was essentially 0；
Production the acid concentration 120mg/g, acid production speed 1.25g/L/h of LCDA；Alkane mass transitions rate 80%.
Claims (10)
1. a kind of method of fermenting and producing long-chain biatomic acid, it is characterised in that: in the fermentation process, rate of producing acid be 1.0~
2.0g/L/h, preferably 1.1~1.7g/L/h, more preferable 1.2~1.5g/L/h.
2. the method as described in claim 1, it is characterised in that: the ventilation quantity in the fermentation process is 0.1~1vvm, preferably
0.2~0.6vvm.
3. method according to claim 1 or 2, it is characterised in that: the strain of the fermentation includes candida tropicalis
(Candida Tropicalis) or candida sake (Candidasake), preferably candida tropicalis (Candida
Tropicalis) (deposit number is CCTCC M203052) or candida tropicalis (Candida Tropicalis)
CATN145 (deposit number is CCTCC M 2011192) or candida sake (Candidasake) CATH4013 (deposit number
For CCTCC M2011486) or candida sake (Candidasake) CATH4014 (deposit number CCTCC
M2011487) perhaps candida sake (Candidasake) CATH4012 (deposit number be CCTCC M2011485) or
Candida sake (Candidasake) CATH4016 (deposit number is CCTCC M2011488) or candida sake
(Candidasake) CATH430 (deposit number is CCTCC M2011489).
4. the method according to claim 1, it is characterised in that: the substrate of the fermentation include C9~C18 just
One of alkane, linear saturated fatty acids, linear saturated fatty acids ester and salts of straight-chain saturated fatty acids are a variety of；It preferably includes
One of normal alkane, linear saturated fatty acids, linear saturated fatty acids ester and salts of straight-chain saturated fatty acids of C11~C16 or
It is a variety of；The more preferably normal alkane of C11, C12, C13, C15, C14 or C16, linear saturated fatty acids, linear saturated fatty acids ester
One of with salts of straight-chain saturated fatty acids.
5. method according to any of claims 1-4, it is characterised in that: the tank temperature when fermentation is 28~32 DEG C.
6. method according to any of claims 1-4, it is characterised in that: tank when fermentation pressure for 0.05~
0.14Mpa, the pressure are gauge pressure.
7. method according to any of claims 1-4, it is characterised in that: the pH when fermentation is 5.0~8.5.
8. method according to any of claims 1-4, it is characterised in that: the fermentation period is 100~180h.
9. method according to any of claims 1-4, it is characterised in that: the speed of agitator when fermentation is 500~
1000rpm, the mixing speed are characterized with the speed of agitator of two layers of turbine disc blade stirring paddle.
10. method according to any of claims 1-4, it is characterised in that: in the fermentation process, the mixing speed and
Ventilation quantity meets the following conditions:
0.6-0.00042A≤F≤0.8-0.00038A；
Wherein: A indicates that the numerical value of mixing speed, the numerical value are the numerical value as unit of rpm；The mixing speed is with two layers of turbine
The speed of agitator of disk blade stirring paddle characterizes；
F indicates that the numerical value of ventilation quantity, the numerical value are the numerical value as unit of vvm.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201711379782.4A CN109943599B (en) | 2017-12-20 | 2017-12-20 | Method for producing long-chain dicarboxylic acid by fermentation |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201711379782.4A CN109943599B (en) | 2017-12-20 | 2017-12-20 | Method for producing long-chain dicarboxylic acid by fermentation |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN109943599A true CN109943599A (en) | 2019-06-28 |
| CN109943599B CN109943599B (en) | 2023-04-14 |
Family
ID=67004801
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201711379782.4A Active CN109943599B (en) | 2017-12-20 | 2017-12-20 | Method for producing long-chain dicarboxylic acid by fermentation |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN109943599B (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2021073011A1 (en) * | 2019-10-18 | 2021-04-22 | 上海凯赛生物技术股份有限公司 | Strain for producing long-chain dicarboxylic acids and fermentation method therefor |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1257126A (en) * | 1998-12-16 | 2000-06-21 | 中国石油化工集团公司 | Process for producing alpha, omega-long chain binary acid by using microorganism fermentation |
| CN1570124A (en) * | 2004-05-12 | 2005-01-26 | 上海凯赛生物技术研发中心有限公司 | Long chain normal dibasic acid production method |
| CN104862348A (en) * | 2014-02-26 | 2015-08-26 | 中国科学院过程工程研究所 | Method for fermentation-membrane separation combined production of long-chain dicarboxylic acid |
| CN104862347A (en) * | 2014-02-26 | 2015-08-26 | 中国科学院过程工程研究所 | Method for producing long-chain binary acid through fermentation separation coupling |
-
2017
- 2017-12-20 CN CN201711379782.4A patent/CN109943599B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1257126A (en) * | 1998-12-16 | 2000-06-21 | 中国石油化工集团公司 | Process for producing alpha, omega-long chain binary acid by using microorganism fermentation |
| CN1570124A (en) * | 2004-05-12 | 2005-01-26 | 上海凯赛生物技术研发中心有限公司 | Long chain normal dibasic acid production method |
| CN104862348A (en) * | 2014-02-26 | 2015-08-26 | 中国科学院过程工程研究所 | Method for fermentation-membrane separation combined production of long-chain dicarboxylic acid |
| CN104862347A (en) * | 2014-02-26 | 2015-08-26 | 中国科学院过程工程研究所 | Method for producing long-chain binary acid through fermentation separation coupling |
Non-Patent Citations (1)
| Title |
|---|
| 王玉亭等: "利用醋酸衍生物发酵制取长链二元酸", 《石油炼制与化工》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2021073011A1 (en) * | 2019-10-18 | 2021-04-22 | 上海凯赛生物技术股份有限公司 | Strain for producing long-chain dicarboxylic acids and fermentation method therefor |
Also Published As
| Publication number | Publication date |
|---|---|
| CN109943599B (en) | 2023-04-14 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Ghose et al. | Rapid ethanol fermentation of cellulose hydrolysate. II. Product and substrate inhibition and optimization of fermentor design | |
| TWI608097B (en) | A microorganism producing lactic acid and a method for producing lactic acid using the same | |
| CN107119002B (en) | Recombinant bacterium for synthesizing 3-hydroxypropionic acid and construction method and application thereof | |
| CN104487582A (en) | Method for preparing organic acid by batch-feeding carbon source substrate and base | |
| CN102076863A (en) | Process for producing organic acid | |
| CN113186147A (en) | Fermentation method for improving production of porcine myoglobin by pichia pastoris engineering bacteria | |
| AU762656B2 (en) | Method of producing gamma-decalactone | |
| CN100363490C (en) | Metabolically engineered micro-organisms having reduced production of undesired metabolic products | |
| CN107815446A (en) | A kind of high cell density fermentation for recombinating nitrile hydratase Recombinant organism | |
| Yang et al. | Production of erythritol from glucose by an osmophilic mutant of Candida magnoliae | |
| CN109943598A (en) | A kind of method of fermenting and producing long-chain biatomic acid | |
| CN101821399A (en) | Method for production of succinic acid | |
| CN109943599A (en) | A kind of method of fermenting and producing long-chain biatomic acid | |
| CN107227286A (en) | Genetically engineered bacterium capable of producing succinic acid at high yield, and construction method and application thereof | |
| CN1952164A (en) | Combined fermentation process for producing D-lactic acid | |
| CN111304138B (en) | Recombinant escherichia coli for producing beta-carotene and construction method and application thereof | |
| CN106755166B (en) | Method for catalytically synthesizing nicotinamide by high molecular weight nitrile hydratase engineering bacteria | |
| CN113293107B (en) | Saccharomyces cerevisiae for industrial production with high organic acid tolerance and construction method thereof | |
| CN114369628A (en) | Method for producing long-chain dicarboxylic acid by biological fermentation | |
| CN111718968A (en) | Method for increasing yield of medium-chain fatty acid of mucor circinelloides WJ11 | |
| Zhou et al. | Influence of glucose feeding on the ligninolytic enzyme production of the white-rot fungus Phanerochaete chrysosporium | |
| CN110951766A (en) | Method for synthesizing L-ornithine by metabolizing mannitol by using recombinant corynebacterium glutamicum | |
| CN106337064B (en) | Production method of L-malic acid | |
| CN109868294A (en) | A method of continuously ferment and produces long-chain biatomic acid | |
| CN114507611B (en) | High-temperature-resistant yeast for producing alpha-ketoglutarate and fermentation method |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| CB02 | Change of applicant information |
Address after: 4 / F, building 5, No. 1690, Cailun Road, Shanghai Free Trade Zone Applicant after: CATHAY R&D CENTER Co.,Ltd. Applicant after: CATHAY INDUSTRIAL BIOTECH Ltd. Address before: 200120 Shanghai Zhangjiang High Tech Park of Pudong New Area Cailun Road No. 5 No. 1690 Applicant before: CATHAY R&D CENTER Co.,Ltd. Applicant before: CATHAY INDUSTRIAL BIOTECH Ltd. |
|
| CB02 | Change of applicant information | ||
| TA01 | Transfer of patent application right |
Effective date of registration: 20191030 Address after: 4 / F, building 5, No. 1690, Cailun Road, Shanghai Free Trade Zone Applicant after: CATHAY R&D CENTER Co.,Ltd. Applicant after: CIBT USA Address before: 201203 floor 4, building 5, No. 1690, Cailun Road, Shanghai pilot Free Trade Zone Applicant before: CATHAY R&D CENTER Co.,Ltd. Applicant before: CATHAY INDUSTRIAL BIOTECH Ltd. |
|
| TA01 | Transfer of patent application right | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| TR01 | Transfer of patent right |
Effective date of registration: 20231130 Address after: 4 / F, building 5, No. 1690, Cailun Road, free trade zone, Pudong New Area, Shanghai Patentee after: CATHAY R&D CENTER Co.,Ltd. Patentee after: CIBT USA Patentee after: Kasai (Shanghai) Biotechnology Co.,Ltd. Address before: 4 / F, building 5, No. 1690, Cailun Road, Shanghai pilot Free Trade Zone, 201203 Patentee before: CATHAY R&D CENTER Co.,Ltd. Patentee before: CIBT USA |
|
| TR01 | Transfer of patent right |