CN115960973A - Undecanedioic acid fermentation liquid, undecanedioic acid fermentation treatment liquid, undecanedioic acid and preparation method thereof - Google Patents
Undecanedioic acid fermentation liquid, undecanedioic acid fermentation treatment liquid, undecanedioic acid and preparation method thereof Download PDFInfo
- Publication number
- CN115960973A CN115960973A CN202111172639.4A CN202111172639A CN115960973A CN 115960973 A CN115960973 A CN 115960973A CN 202111172639 A CN202111172639 A CN 202111172639A CN 115960973 A CN115960973 A CN 115960973A
- Authority
- CN
- China
- Prior art keywords
- fermentation
- acid
- undecanedioic acid
- liquid
- undecane
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 238000000855 fermentation Methods 0.000 title claims abstract description 246
- 230000004151 fermentation Effects 0.000 title claims abstract description 246
- LWBHHRRTOZQPDM-UHFFFAOYSA-N undecanedioic acid Chemical compound OC(=O)CCCCCCCCCC(O)=O LWBHHRRTOZQPDM-UHFFFAOYSA-N 0.000 title claims abstract description 142
- 239000007788 liquid Substances 0.000 title claims abstract description 73
- 238000011282 treatment Methods 0.000 title claims abstract description 44
- 238000002360 preparation method Methods 0.000 title claims abstract description 8
- RSJKGSCJYJTIGS-UHFFFAOYSA-N N-undecane Natural products CCCCCCCCCCC RSJKGSCJYJTIGS-UHFFFAOYSA-N 0.000 claims abstract description 94
- 238000000034 method Methods 0.000 claims abstract description 60
- -1 undecane alkane Chemical class 0.000 claims abstract description 26
- 238000000746 purification Methods 0.000 claims abstract description 13
- 240000004808 Saccharomyces cerevisiae Species 0.000 claims abstract description 12
- 238000011218 seed culture Methods 0.000 claims abstract description 12
- 235000002639 sodium chloride Nutrition 0.000 claims description 45
- 150000003839 salts Chemical class 0.000 claims description 43
- 239000002253 acid Substances 0.000 claims description 39
- 239000007787 solid Substances 0.000 claims description 31
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 claims description 27
- 229930195733 hydrocarbon Natural products 0.000 claims description 22
- 241000222178 Candida tropicalis Species 0.000 claims description 21
- 239000004215 Carbon black (E152) Substances 0.000 claims description 21
- 241001530515 Candida sake Species 0.000 claims description 17
- 229910052799 carbon Inorganic materials 0.000 claims description 17
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 claims description 15
- 230000008569 process Effects 0.000 claims description 15
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 14
- 241000222120 Candida <Saccharomycetales> Species 0.000 claims description 13
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 claims description 12
- XSQUKJJJFZCRTK-UHFFFAOYSA-N Urea Chemical compound NC(N)=O XSQUKJJJFZCRTK-UHFFFAOYSA-N 0.000 claims description 12
- 239000001963 growth medium Substances 0.000 claims description 12
- FGIUAXJPYTZDNR-UHFFFAOYSA-N potassium nitrate Chemical compound [K+].[O-][N+]([O-])=O FGIUAXJPYTZDNR-UHFFFAOYSA-N 0.000 claims description 12
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims description 11
- 238000000605 extraction Methods 0.000 claims description 11
- 239000008103 glucose Substances 0.000 claims description 11
- 230000001580 bacterial effect Effects 0.000 claims description 10
- 238000002425 crystallisation Methods 0.000 claims description 10
- 230000008025 crystallization Effects 0.000 claims description 10
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical group CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 claims description 9
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical group CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 claims description 9
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 9
- 238000000926 separation method Methods 0.000 claims description 9
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 8
- 239000002518 antifoaming agent Substances 0.000 claims description 8
- 238000001816 cooling Methods 0.000 claims description 8
- 239000003960 organic solvent Substances 0.000 claims description 8
- 230000001502 supplementing effect Effects 0.000 claims description 7
- 241000222157 Candida viswanathii Species 0.000 claims description 6
- KFZMGEQAYNKOFK-UHFFFAOYSA-N Isopropanol Chemical compound CC(C)O KFZMGEQAYNKOFK-UHFFFAOYSA-N 0.000 claims description 6
- LRHPLDYGYMQRHN-UHFFFAOYSA-N N-Butanol Chemical compound CCCCO LRHPLDYGYMQRHN-UHFFFAOYSA-N 0.000 claims description 6
- 239000004721 Polyphenylene oxide Substances 0.000 claims description 6
- 240000008042 Zea mays Species 0.000 claims description 6
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 claims description 6
- 235000002017 Zea mays subsp mays Nutrition 0.000 claims description 6
- 239000004202 carbamide Substances 0.000 claims description 6
- 235000013877 carbamide Nutrition 0.000 claims description 6
- 235000005822 corn Nutrition 0.000 claims description 6
- 150000002148 esters Chemical class 0.000 claims description 6
- 229910017053 inorganic salt Inorganic materials 0.000 claims description 6
- 150000002576 ketones Chemical class 0.000 claims description 6
- 229910000402 monopotassium phosphate Inorganic materials 0.000 claims description 6
- 235000019796 monopotassium phosphate Nutrition 0.000 claims description 6
- 229910052757 nitrogen Inorganic materials 0.000 claims description 6
- 239000002245 particle Substances 0.000 claims description 6
- 229920000570 polyether Polymers 0.000 claims description 6
- 239000004323 potassium nitrate Substances 0.000 claims description 6
- 235000010333 potassium nitrate Nutrition 0.000 claims description 6
- 229940041514 candida albicans extract Drugs 0.000 claims description 5
- 238000011081 inoculation Methods 0.000 claims description 5
- 239000012138 yeast extract Substances 0.000 claims description 5
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 claims description 4
- 229930006000 Sucrose Natural products 0.000 claims description 4
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 claims description 4
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 claims description 4
- 229910052921 ammonium sulfate Inorganic materials 0.000 claims description 4
- 235000011130 ammonium sulphate Nutrition 0.000 claims description 4
- 238000006243 chemical reaction Methods 0.000 claims description 4
- WRUGWIBCXHJTDG-UHFFFAOYSA-L magnesium sulfate heptahydrate Chemical compound O.O.O.O.O.O.O.[Mg+2].[O-]S([O-])(=O)=O WRUGWIBCXHJTDG-UHFFFAOYSA-L 0.000 claims description 4
- 229940061634 magnesium sulfate heptahydrate Drugs 0.000 claims description 4
- 239000011780 sodium chloride Substances 0.000 claims description 4
- DKPFZGUDAPQIHT-UHFFFAOYSA-N Butyl acetate Natural products CCCCOC(C)=O DKPFZGUDAPQIHT-UHFFFAOYSA-N 0.000 claims description 3
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 claims description 3
- FUZZWVXGSFPDMH-UHFFFAOYSA-N hexanoic acid Chemical compound CCCCCC(O)=O FUZZWVXGSFPDMH-UHFFFAOYSA-N 0.000 claims description 3
- 239000001301 oxygen Substances 0.000 claims description 3
- 229910052760 oxygen Inorganic materials 0.000 claims description 3
- 230000020477 pH reduction Effects 0.000 claims description 3
- 239000000843 powder Substances 0.000 claims description 3
- 239000005720 sucrose Substances 0.000 claims 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 claims 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 claims description 2
- 150000007513 acids Chemical class 0.000 claims description 2
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 claims description 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 claims description 2
- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 claims description 2
- 235000003891 ferrous sulphate Nutrition 0.000 claims description 2
- 239000011790 ferrous sulphate Substances 0.000 claims description 2
- BAUYGSIQEAFULO-UHFFFAOYSA-L iron(2+) sulfate (anhydrous) Chemical compound [Fe+2].[O-]S([O-])(=O)=O BAUYGSIQEAFULO-UHFFFAOYSA-L 0.000 claims description 2
- 229910000359 iron(II) sulfate Inorganic materials 0.000 claims description 2
- 229910052943 magnesium sulfate Inorganic materials 0.000 claims description 2
- 235000019341 magnesium sulphate Nutrition 0.000 claims description 2
- 229910000403 monosodium phosphate Inorganic materials 0.000 claims description 2
- 235000019799 monosodium phosphate Nutrition 0.000 claims description 2
- PJNZPQUBCPKICU-UHFFFAOYSA-N phosphoric acid;potassium Chemical compound [K].OP(O)(O)=O PJNZPQUBCPKICU-UHFFFAOYSA-N 0.000 claims description 2
- 229920001296 polysiloxane Polymers 0.000 claims description 2
- AJPJDKMHJJGVTQ-UHFFFAOYSA-M sodium dihydrogen phosphate Chemical compound [Na+].OP(O)([O-])=O AJPJDKMHJJGVTQ-UHFFFAOYSA-M 0.000 claims description 2
- 238000004042 decolorization Methods 0.000 claims 2
- 239000007789 gas Substances 0.000 claims 2
- 150000001298 alcohols Chemical class 0.000 claims 1
- 238000004519 manufacturing process Methods 0.000 abstract description 18
- HCUZVMHXDRSBKX-UHFFFAOYSA-N 2-decylpropanedioic acid Chemical compound CCCCCCCCCCC(C(O)=O)C(O)=O HCUZVMHXDRSBKX-UHFFFAOYSA-N 0.000 abstract description 5
- 230000007613 environmental effect Effects 0.000 abstract 1
- 235000010633 broth Nutrition 0.000 description 31
- 239000000758 substrate Substances 0.000 description 16
- 239000000243 solution Substances 0.000 description 14
- ZDPHROOEEOARMN-UHFFFAOYSA-N undecanoic acid Chemical compound CCCCCCCCCCC(O)=O ZDPHROOEEOARMN-UHFFFAOYSA-N 0.000 description 14
- 238000004321 preservation Methods 0.000 description 9
- 150000001335 aliphatic alkanes Chemical group 0.000 description 8
- 238000001704 evaporation Methods 0.000 description 8
- 241001052560 Thallis Species 0.000 description 7
- 230000004913 activation Effects 0.000 description 7
- 230000002035 prolonged effect Effects 0.000 description 7
- 239000000126 substance Substances 0.000 description 7
- 239000003513 alkali Substances 0.000 description 6
- 230000000052 comparative effect Effects 0.000 description 6
- 238000012258 culturing Methods 0.000 description 5
- 239000013530 defoamer Substances 0.000 description 5
- 239000002609 medium Substances 0.000 description 5
- 239000000047 product Substances 0.000 description 5
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 5
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 4
- MHAJPDPJQMAIIY-UHFFFAOYSA-N Hydrogen peroxide Chemical compound OO MHAJPDPJQMAIIY-UHFFFAOYSA-N 0.000 description 4
- 238000001035 drying Methods 0.000 description 4
- 230000000694 effects Effects 0.000 description 4
- 238000001914 filtration Methods 0.000 description 4
- GNSKLFRGEWLPPA-UHFFFAOYSA-M potassium dihydrogen phosphate Chemical compound [K+].OP(O)([O-])=O GNSKLFRGEWLPPA-UHFFFAOYSA-M 0.000 description 4
- LWIHDJKSTIGBAC-UHFFFAOYSA-K potassium phosphate Substances [K+].[K+].[K+].[O-]P([O-])([O-])=O LWIHDJKSTIGBAC-UHFFFAOYSA-K 0.000 description 4
- 239000013078 crystal Substances 0.000 description 3
- 230000008020 evaporation Effects 0.000 description 3
- 239000012535 impurity Substances 0.000 description 3
- 150000004668 long chain fatty acids Chemical class 0.000 description 3
- 230000003287 optical effect Effects 0.000 description 3
- 239000002994 raw material Substances 0.000 description 3
- 159000000000 sodium salts Chemical class 0.000 description 3
- 229960004793 sucrose Drugs 0.000 description 3
- 238000003786 synthesis reaction Methods 0.000 description 3
- 241000894006 Bacteria Species 0.000 description 2
- BVKZGUZCCUSVTD-UHFFFAOYSA-L Carbonate Chemical compound [O-]C([O-])=O BVKZGUZCCUSVTD-UHFFFAOYSA-L 0.000 description 2
- 101000631695 Homo sapiens Succinate dehydrogenase assembly factor 3, mitochondrial Proteins 0.000 description 2
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 2
- OFOBLEOULBTSOW-UHFFFAOYSA-N Malonic acid Chemical compound OC(=O)CC(O)=O OFOBLEOULBTSOW-UHFFFAOYSA-N 0.000 description 2
- 229910002651 NO3 Inorganic materials 0.000 description 2
- NHNBFGGVMKEFGY-UHFFFAOYSA-N Nitrate Chemical compound [O-][N+]([O-])=O NHNBFGGVMKEFGY-UHFFFAOYSA-N 0.000 description 2
- 229910019142 PO4 Inorganic materials 0.000 description 2
- 102100028996 Succinate dehydrogenase assembly factor 3, mitochondrial Human genes 0.000 description 2
- QAOWNCQODCNURD-UHFFFAOYSA-L Sulfate Chemical compound [O-]S([O-])(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-L 0.000 description 2
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 2
- 150000003863 ammonium salts Chemical class 0.000 description 2
- 159000000007 calcium salts Chemical class 0.000 description 2
- 230000010261 cell growth Effects 0.000 description 2
- 238000012136 culture method Methods 0.000 description 2
- 230000007423 decrease Effects 0.000 description 2
- 238000001514 detection method Methods 0.000 description 2
- 238000010790 dilution Methods 0.000 description 2
- 239000012895 dilution Substances 0.000 description 2
- 239000003814 drug Substances 0.000 description 2
- 239000011521 glass Substances 0.000 description 2
- 235000011187 glycerol Nutrition 0.000 description 2
- 230000012010 growth Effects 0.000 description 2
- 150000002430 hydrocarbons Chemical class 0.000 description 2
- 150000002505 iron Chemical class 0.000 description 2
- 159000000003 magnesium salts Chemical class 0.000 description 2
- 239000012528 membrane Substances 0.000 description 2
- 230000002503 metabolic effect Effects 0.000 description 2
- 244000005700 microbiome Species 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 description 2
- 239000010452 phosphate Substances 0.000 description 2
- 238000006116 polymerization reaction Methods 0.000 description 2
- XAEFZNCEHLXOMS-UHFFFAOYSA-M potassium benzoate Chemical compound [K+].[O-]C(=O)C1=CC=CC=C1 XAEFZNCEHLXOMS-UHFFFAOYSA-M 0.000 description 2
- 239000006228 supernatant Substances 0.000 description 2
- 238000005406 washing Methods 0.000 description 2
- 238000005303 weighing Methods 0.000 description 2
- 108090000790 Enzymes Proteins 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- ALHUZKCOMYUFRB-OAHLLOKOSA-N Muscone Chemical compound C[C@@H]1CCCCCCCCCCCCC(=O)C1 ALHUZKCOMYUFRB-OAHLLOKOSA-N 0.000 description 1
- 230000001133 acceleration Effects 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 230000003213 activating effect Effects 0.000 description 1
- 125000001931 aliphatic group Chemical group 0.000 description 1
- 125000004432 carbon atom Chemical group C* 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 230000003247 decreasing effect Effects 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 230000007123 defense Effects 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 238000005265 energy consumption Methods 0.000 description 1
- 229920006351 engineering plastic Polymers 0.000 description 1
- 230000006353 environmental stress Effects 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 230000002349 favourable effect Effects 0.000 description 1
- 238000004817 gas chromatography Methods 0.000 description 1
- 239000004615 ingredient Substances 0.000 description 1
- 239000000543 intermediate Substances 0.000 description 1
- 239000000314 lubricant Substances 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 238000005374 membrane filtration Methods 0.000 description 1
- 125000002496 methyl group Chemical group [H]C([H])([H])* 0.000 description 1
- 238000001471 micro-filtration Methods 0.000 description 1
- 239000011259 mixed solution Substances 0.000 description 1
- ALHUZKCOMYUFRB-UHFFFAOYSA-N muskone Natural products CC1CCCCCCCCCCCCC(=O)C1 ALHUZKCOMYUFRB-UHFFFAOYSA-N 0.000 description 1
- 238000010899 nucleation Methods 0.000 description 1
- 230000003647 oxidation Effects 0.000 description 1
- 238000007254 oxidation reaction Methods 0.000 description 1
- 239000002304 perfume Substances 0.000 description 1
- 239000000575 pesticide Substances 0.000 description 1
- 239000012450 pharmaceutical intermediate Substances 0.000 description 1
- 239000008020 pharmaceutical preservative Substances 0.000 description 1
- 230000008092 positive effect Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 239000011347 resin Substances 0.000 description 1
- 229920005989 resin Polymers 0.000 description 1
- 239000010865 sewage Substances 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 230000001954 sterilising effect Effects 0.000 description 1
- 230000002194 synthesizing effect Effects 0.000 description 1
- 229920002994 synthetic fiber Polymers 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 230000009466 transformation Effects 0.000 description 1
- 238000000108 ultra-filtration Methods 0.000 description 1
Classifications
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02E—REDUCTION OF GREENHOUSE GAS [GHG] EMISSIONS, RELATED TO ENERGY GENERATION, TRANSMISSION OR DISTRIBUTION
- Y02E50/00—Technologies for the production of fuel of non-fossil origin
- Y02E50/10—Biofuels, e.g. bio-diesel
Landscapes
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
The invention relates to the technical field of fermentation, and discloses a undecanedioic acid fermentation liquid, a undecanedioic acid fermentation treatment liquid, an undecanedioic acid and a preparation method thereof, wherein the method comprises the following steps: performing seed culture and fermentation culture on yeast, adding undecane alkane during the fermentation culture, and controlling pH of the fermentation broth to be below 7.0, further 4.5-7.0 to obtain undecane dicarboxylic acid fermentation broth. The method provided by the invention can obviously improve the yield of the undecanedioic acid and shorten the fermentation time, and simultaneously can obviously lighten the subsequent purification process, thereby reducing the production cost and the environmental pressure.
Description
Technical Field
The invention relates to the technical field of fermentation, in particular to a undecanedioic acid fermentation liquid, a undecanedioic acid fermentation treatment liquid, an undecanedioic acid and a preparation method thereof.
Background
The long-chain dibasic acid (DCA) is aliphatic dibasic acid with 10 or more than 10 carbon atoms in a carbon chain, is a chemical product with very important and wide application and does not exist independently in the natural world. Because the chain segment contains long alkane chain segments, the long alkane chain segments have the property superior to short chain dibasic acid, so that the corresponding synthetic material has superior performance. Among them, undecanedioic acid (α, ω -dicarboxy long chain dibasic acid, abbreviated as DC 11) is a common important dibasic acid, and can be used for synthesizing perfumes, pharmaceutical intermediates, preservatives, and the like. And the derivative product of the eleven-carbon dicarboxylic acid is an important raw material or intermediate for manufacturing muscone, medicine, resin, pesticide, high-grade engineering plastic, high-grade lubricant and the like. Therefore, the method is widely applied to the fields of national defense, light industry, chemical industry, medicine, automobile industry, engineering materials and the like.
At present, the eleven-dicarboxylic acid at home and abroad is mostly synthesized by a chemical synthesis method or a biological fermentation method. However, the traditional chemical synthesis method has the disadvantages of long synthetic route, high equipment requirement, low yield, high energy consumption, serious pollution and the like in the production process. The biological fermentation method is simple, and takes n-undecane hydrocarbon as a substrate, and can oxidize methyl at two ends of the n-undecane molecular chain under normal temperature and pressure to generate undecane diacid by the alpha and omega oxidation ability of microorganism and the action of related enzymes in the microorganism cell. Compared with the chemical synthesis method, the biological fermentation method has the advantages of simple fermentation process, lower pollution, low cost and the like.
The research work of the long-chain dicarboxylic acid has been carried out for more than 40 years in China, and the report of producing the undecane dicarboxylic acid by a biological fermentation method is few, so that the development of a novel production process of the undecane dicarboxylic acid has very important significance.
Disclosure of Invention
In order to overcome the defects in the existing undecanedioic acid production process, the invention provides an undecanedioic acid fermentation liquid, an undecanedioic acid fermentation treatment liquid, an undecanedioic acid and a preparation method thereof.
In order to solve the above problems, a first aspect of the present invention provides a method for preparing a undecanoic diacid fermentation broth, comprising: performing seed culture and fermentation culture on yeast, adding undecane hydrocarbon during the fermentation culture process, and controlling pH of the fermentation liquid to be below 7.0, further 4.5-7.0 to obtain undecane diacid fermentation liquid.
Preferably, the pH of the fermentation broth is controlled to a value of 5.0 to 7.0, preferably 5.5 to 6.5.
Preferably, the concentration of undecane in the fermentation broth is controlled to be 0.1-5% (w/v), preferably 1-3% (w/v) during the fermentation culture.
Preferably, the concentration of viable cells in the fermentation broth is controlled to 3 x 10 during the fermentation culture 7 cfu/mL-4*10 9 cfu/mL, preferably 2 x 10 8 cfu/mL-8*10 8 cfu/mL。
In a second aspect, the present invention provides a method for preparing undecanedioic acid, which comprises: according to the method of the first aspect of the invention, undecanedioic acid fermentation liquor is obtained, and then the undecanedioic acid fermentation liquor is extracted and purified, or the undecanedioic acid fermentation liquor is processed to obtain fermentation treatment liquid, and then the fermentation treatment liquid is extracted and purified; to obtain the undecanedioic acid.
In a third aspect, the present invention provides an undecanedioic acid prepared by the process of the second aspect of the present invention.
In a fourth aspect, the present invention provides a undecanedioic acid fermentation broth, wherein the salt content in the undecanedioic acid fermentation broth is below 15%, preferably below 10%, more preferably below 8%; the percentage is the mass percentage of the total amount of the undecanedioic acid generated by fermentation.
The fifth aspect of the invention provides a undecanedioic acid fermentation treatment liquid, wherein the salt content in the fermentation treatment liquid is 8000ppm or less, preferably 6000ppm or less, and the mass per million of the salt is the mass per million of the undecanedioic acid fermentation treatment liquid; the fermentation treatment liquid is obtained by acidifying the undecane dibasic acid fermentation liquid of the fourth aspect of the invention and removing solid matter; the solids comprise particles of undecanedioic acid.
Compared with the prior art, the invention has the following positive effects:
the method provided by the invention can enable the yield of the undecanedioic acid to reach more than 160g/L, and in addition, the fermentation period is greatly shortened, and the fermentation time can be controlled to be about 160h, so that the production efficiency is improved, and the production cost of the undecanedioic acid is reduced.
Particularly, the fermentation method can effectively reduce the salt content generated in the production process of the undecane diacid, not only saves the cost of raw materials, but also can effectively reduce the downstream pollution, and also obviously reduces the cost of sewage treatment, thereby further reducing the production cost of the undecane diacid.
Detailed Description
The endpoints of the ranges and any values disclosed herein are not limited to the precise range or value, and such ranges or values should be understood to encompass values close to those ranges or values. For numerical ranges, each range between its endpoints and individual point values, and each individual point value can be combined with each other to give one or more new numerical ranges, and such numerical ranges should be construed as specifically disclosed herein.
As described above, the first aspect of the present invention provides a method for preparing a undecanoic acid fermentation broth, comprising: performing seed culture and fermentation culture on yeast, adding undecane alkane during the fermentation culture, and controlling pH of the fermentation broth to be below 7.0, further 4.5-7.0 to obtain undecane dicarboxylic acid fermentation broth.
According to a preferred embodiment of the invention, the pH of the fermentation broth is controlled to a pH of from 5.0 to 6.5, preferably from 5.5 to 6.5. Specific examples of pH values may be: 4.5,4.6,4.7,4.8,4.9,5.0,5.1,5.2,5.3,5.4,5.5,5.6,5.7,5.8,5.9,6.0,6.1,6.2,6.3,6.4,6.5,6.6,6.7,6.8,6.9,7.0.
In some embodiments of the invention, the OD of the bacterial cells is 620 When the value reaches more than 0.6-1.0 (diluted by 30 times), adding undecane alkane into the fermentation liquid, and producing a large amount of undecane dicarboxylic acid after the thallus enters the conversion stage。
In the present invention, the undecakane may be added by batch addition or continuous feed, so that the concentration of the substrate undecakane in the fermentation broth is 0.1-5% (w/v), and preferably the concentration of the fermentation substrate is controlled within any one of the ranges of 0.1-0.5% (w/v), 0.5-1.0% (w/v), 1.0-2.0% (w/v), 2.0-3.0% (w/v), 3.0-4.0% (w/v), 4.0-5.0% (w/v), or 1.0-3.0% (w/v), or any combination of these ranges, such as a concentration range of 0.5-2.0% (w/v) between 0.5% and 2.0%, or a concentration range of 3.0-5.0% (w/v) between 3.0% and 5.0%. In the present invention, the percentage% (w/v) represents a mass-to-volume ratio, i.e., g/100mL.
According to a preferred embodiment of the present invention, when the undecane is fed by continuous feeding, the feeding rate of the undecane is preferably 0.01 to 0.5L/h, more preferably 0.01 to 0.08L/h, further preferably 0.02 to 0.08L/h. In such a preferable case, the amount of the acid produced can be further increased and the fermentation time can be reduced.
In some specific embodiments of the invention, the temperature is controlled to be 26-35 ℃, preferably 28-32 ℃ during the fermentation culture process; the air volume is 0.1-1.0vvm, preferably 0.1-0.6vvm, more preferably 0.2-0.4vvm; the pressure is 0.05 to 0.15MPa, preferably 0.08 to 0.12MPa.
In some embodiments of the invention, dissolved oxygen is controlled to be above 15%, preferably 20-40%, such as 25%, 30%, 35%, etc., during the fermentation culture.
In some specific embodiments of the invention, the concentration of viable cells in the fermentation broth is controlled to 3 x 10 during the fermentation culture 7 cfu/mL-4*10 9 cfu/mL, preferably 2 x 10 8 cfu/mL-8*10 8 cfu/mL. In particular, the concentration of viable cells can be controlled by controlling the rate of substrate flow. The inventor finds that the pH value is above 7, the cell growth is not facilitated, the online viable cell concentration and the OD value are lower in the fermentation process, the fermentation period is prolonged, and the DC11 yield is lower; the pH value below 7 is favorable for cell growth, the online viable cell concentration is higher in the fermentation process, the fermentation period is short, and DC11 the yield is higher.
In some specific embodiments of the invention, when seed culture and fermentation culture are performed, the seed tank medium and the fermentor medium used include a carbon source, a nitrogen source, inorganic salts, and an antifoaming agent. Wherein, the carbon source includes but is not limited to one or more of glucose, sucrose and maltose; and/or, the nitrogen source includes, but is not limited to, one or more of yeast extract, ammonium sulfate, urea, corn steep liquor, potassium nitrate, and yeast powder; and/or, the inorganic salt includes but is not limited to one or more of magnesium sulfate heptahydrate, potassium nitrate, ferrous sulfate, sodium chloride, dipotassium hydrogen phosphate, magnesium sulfate, sodium dihydrogen phosphate, sodium chloride and potassium dihydrogen phosphate; and/or, the defoamer includes, but is not limited to, a silicone type defoamer and/or a polyether type defoamer. The addition amounts of the carbon source, nitrogen source, inorganic salt and defoaming agent are not particularly limited in the present invention, and may be selected conventionally in the art.
In some specific embodiments of the invention, the seedtank medium includes the following ingredients: 1.0 to 3.2 percent of cane sugar, 0.1 to 0.8 percent of urea, 0.1 to 1.5 percent of monopotassium phosphate, 0.1 to 0.6 percent of yeast powder, 0.1 to 0.5 percent of corn steep liquor and 0.02 to 0.10 percent of defoaming agent (polyether). In the present invention, the percentages represent mass to volume (w/v), i.e.% means g/100mL, for example, a urea content of 0.1% to 0.8% means a urea content of 0.1g/100mL to 0.8g/100mL.
In some embodiments of the invention, the fermentor medium comprises the following components: 1.0 to 6.0 percent of glucose, 0.1 to 0.6 percent of ammonium sulfate, 0.1 to 0.5 percent of magnesium sulfate heptahydrate, 0.1 to 1.0 percent of monopotassium phosphate, 0.1 to 0.6 percent of yeast extract, 0.1 to 0.6 percent of potassium nitrate, 0.1 to 0.6 percent of corn steep liquor and 0.05 to 0.1 percent of defoaming agent (polyether). In the present invention, the percentage represents a mass-to-volume ratio (w/v), i.e.,% represents g/100mL, for example, glucose content of 1.0% to 6.0% represents 1.0g/100mL to 6.0g/100mL.
In some embodiments of the invention, the temperature is controlled to be 26-32 ℃, preferably 28-30 ℃ during the seed culture; the air volume is 0.1-0.8vvm, preferably 0.4-0.6vvm; the pressure is 0.05-0.15MPa, preferably 0.08-0.12MPa.
In some embodiments of the invention, the seed liquid OD is 620 When the value is more than 0.6 (diluted by 30 times), inoculating seed liquid into a fermentation culture medium for fermentation culture, wherein the inoculation amount of the seed liquid is 5-35% (v/v), preferably 10-30% (v/v). In the invention, the inoculation amount is 10-30% (v/v), which means that the volume of the mature seed liquid inoculated into the fermentation culture medium accounts for 10-30% of the total volume of the fermentation culture medium after the mature seed liquid is inoculated.
In some embodiments of the invention, the yeast comprises Candida Tropicalis (Candida Tropicalis), candida sake (Candida sake) or Candida virginiana (Candida viswanathii). The Candida Tropicalis (Candida Tropicalis), candida sake (Candida sake) or Candida virginiana (Candida viswanathii) used in the present invention are not particularly limited, and are commercially available or can be prepared by itself.
In a preferred embodiment of the present invention, the Candida Tropicalis is preferably Candida Tropicalis (with a preservation number of CCTCC M203052), candida Tropicalis (with a preservation number of CCTCC M2011192), candida Tropicalis (with a preservation number of CCTCC M2013143), or Candida Tropicalis (with a preservation number of CCTCC M2015314), or Candida Tropicalis (with a preservation number of CCTCC M201531303).
In a preferred technical scheme of the present invention, the candida sake is preferably candida sake (candidas) CATH4013 (with a collection number of CCTCC M2011486), or candida sake (candidas) CATH4014 (with a collection number of CCTCC M2011487), or candida sake (candidas) CATH4012 (with a collection number of CCTCC M2011485), or candida sake (candidas) CATH4016 (with a collection number of CCTCC M2011488), or candida sake (candidas) CATH430 (with a collection number of CCTCC M2011489).
In a preferred embodiment of the present invention, the Candida virustata is preferably Candida virustata (Candida viswanathii), and the preservation number is CCTCC NO: m2020048.
The expanding culture method of Candida Tropicalis (Candida Tropicalis), candida visvanica (Candida viswanathii) and Candida sake (Candida sake) is not particularly limited, and a conventional slant strain shaking flask activation and seeding tank culture method can be adopted.
In the invention, before the yeast is subjected to seed culture, strain activation culture is also included, namely, firstly, the yeast is subjected to activation culture in a shake flask, and the OD (optical density) value of the shake flask seeds after being diluted by 30 times is obtained 620 When the concentration reaches above 0.6, inoculating the shake flask seeds into a seed tank filled with a seed culture medium, and culturing until the optical density value OD of the obtained seed liquid is 30 times diluted 620 Up to over 0.6.
Specifically, the pH of 10 Baume wort was adjusted to about 5.4, 80mL of the wort was sterilized in a 500mL Erlenmeyer flask at 121 ℃ for 20min, 1 yeast seed was inoculated into 2mL of glycerin tube stored in a-80 ℃ freezer, and the cells were activated and cultured on a rotary shaker until the OD was 30-fold diluted 620 The value reaches above 0.6, and the activation culture is finished. The conditions for the activation culture of the strain can be specifically as follows: the temperature is 28-32 ℃, and the rotating speed of the shaking table is 150-300rpm. OD 30-fold dilution of the cells after culturing for about 38-44h 620 The value can reach more than 0.6, and the activation culture is finished.
Inoculating the seed in the shake flask into a seed tank containing seed culture medium, typically, when the seed amount in the shake flask is 1.0% -2.0%, such as 1.2% -1.5%, culturing the seed under the above conditions for about 20-24h until the optical density OD of the obtained seed solution after 30 times dilution is reached 620 And (3) reaching above 0.6, and then inoculating the seed liquid into a fermentation culture medium for fermentation and transformation.
According to a preferred embodiment of the invention, the method further comprises: in the fermentation culture process, a primary carbon source is supplemented into a fermentation system. The method for supplementing the primary carbon source is not particularly limited, and can be used for supplementing in batches or continuously in a flow mode, and the method can be selected by a person skilled in the art according to actual conditions.
Preferably, the sugar concentration during fermentation is controlled to be 0.01-5% (w/v), preferably 0.1-0.8% (w/v), more preferably (0.5-0.8)% (w/v), e.g.0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, by supplementing the primary carbon source. In accordance with common knowledge in the field of fermentation, the percentages stated in the present invention are mass to volume ratios, i.e., (w/v) means g/100mL.
Preferably, the concentration of the primary carbon source is 20-70% (w/v), more preferably 40-50% (w/v).
Preferably, the time period for feeding the primary carbon source to the fermentation broth is 5-14h, more preferably 6-11h.
In the present invention, the primary carbon source is preferably glucose.
In a second aspect, the present invention provides a method for preparing undecanedioic acid, which comprises: the undecane diacid fermentation liquor is obtained according to the method, and then the undecane diacid fermentation liquor is extracted and purified, or the undecane diacid fermentation liquor is processed to obtain fermentation treatment liquor, and then the extraction and purification are carried out; to obtain the undecanedioic acid.
In some embodiments of the invention, the method of extraction and purification comprises: a decoloring step;
preferably, the decoloring step comprises decoloring by using activated carbon, and the added amount of the activated carbon is not more than 5% of the volume of the clear liquid; the decolorizing temperature is 80-105 deg.C, and the time is 10-170min.
In some embodiments of the invention, the method of extraction and purification comprises: forming solid matter and separating the solid matter; preferably, the method for forming the solid is acidification crystallization; the separation steps of forming solids and solids may further include any other extraction and purification steps.
In some embodiments of the invention, the decolorizing step comprises: decoloring fermentation liquor or fermentation treatment liquor, or decoloring solution formed by dissolving the solid by an organic solvent; the organic solvent is selected from one or more of alcohol, acid, ketone and ester, wherein the alcohol is selected from one or more of methanol, ethanol, isopropanol and n-butanol; the acid is acetic acid; the ketone is acetone; the ester is ethyl acetate and/or butyl acetate.
In some embodiments of the invention, the method of extraction and purification comprises: forming a solid, dissolving the solid by an organic solvent, separating to obtain a clear liquid, and finally crystallizing and separating the clear liquid to obtain the undecanedioic acid. In the present invention, the organic solvent is selected as described above, and is not described herein again.
In some embodiments of the present invention, the crystallization is a temperature-decreasing crystallization, and the temperature-decreasing crystallization comprises the following steps: cooling to 60-85 deg.C, maintaining for 1-2 hr, cooling to 20-40 deg.C, and crystallizing.
The separation method is not particularly limited in the present invention, and any separation method that is conventional in the art may be used, for example, but not limited to, membrane filtration (microfiltration membrane, ultrafiltration membrane, etc.), plate-and-frame filtration, and centrifugal separation, to remove the bacterial cells, residual hydrocarbons, and metabolic impurities that affect downstream polymerization in the fermentation broth, and to obtain solid substances. The invention preferably adopts a plate-and-frame filtration mode to separate and obtain clear liquid. The present invention preferably adopts a centrifugal separation mode to obtain the undecane dibasic acid.
In the invention, after the crystallization and separation of the clear liquid, the method further comprises the steps of carrying out solvent washing, water washing and drying treatment on the obtained crystal, and then obtaining the undecane diacid product.
Accordingly, in a third aspect, the present invention provides an undecanedioic acid prepared by the process of the second aspect of the present invention.
In the preparation method of the invention, the method for testing the content of the undecane diacid in the fermentation liquor is as follows: in order to determine whether the fermentation liquor contains impurities such as the undecanoic monoacid and other long-chain fatty acids, the contents of the undecanoic acid, the dibasic acid and other long-chain fatty acids in the fermentation liquor can be measured by GC detection through a gas chromatography, namely, the fermentation liquor is subjected to a series of treatments and then enters a capillary column for separation, and the peak-off times of the undecanoic acid, the dibasic acid and other long-chain fatty acid standard products are different.
In a fourth aspect, the present invention provides a undecanedioic acid fermentation broth, wherein the salt content in the undecanedioic acid fermentation broth is below 15%, preferably below 10%, more preferably below 8%; the percentage is the mass percentage relative to the total amount of undecanedioic acid produced by fermentation.
In a preferred embodiment of the present invention, the undecanoic acid fermentation broth is obtained by fermentative conversion at a pH of 7.0 or less, preferably 4.5 to 7.0, more preferably 5.0 to 7.0, and even more preferably 5.5 to 6.5.
In the present invention, the salt in the fermentation liquid includes inorganic salts and salts of undecanedioic acid. Wherein, the inorganic salt accounts for most of the total amount, and the salt of the dibasic acid accounts for a very small amount. The salts of the diacids are generally present in the form of the sodium salts of the diacids.
In the present invention, the salts include, but are not limited to: one or more of potassium salt, sodium salt, magnesium salt, calcium salt, iron salt, ammonium salt, hydrochloride, carbonate, sulfate, nitrate and phosphate.
According to a preferred technical scheme of the invention, the salt content in the fermentation liquor can be 15%,14%,13%,12%,11%,10%,9%,8%,7%,6%,5%,4%,3%,2%,1%,0.8%,0.5%,0.3%; the percentage is the mass percentage of the total amount of the undecanedioic acid generated by fermentation.
The total amount of undecanedioic acid produced by fermentation herein includes undecanedioic acid crystals, according to common general knowledge in the art; or it may also comprise undecanedioic acid precipitated in an amorphous state; it is also possible to include very little undecameric acid precipitated in salt form or very little undecameric acid present in solution in dissolved form in salt form.
The fifth aspect of the invention provides a undecanedioic acid fermentation treatment liquid, wherein the fermentation treatment liquid contains less than 8000ppm of salt, preferably less than 6000ppm of salt, and the parts per million is the mass parts per million of the salt in the undecanedioic acid fermentation treatment liquid; the fermentation treatment liquid is obtained by acidifying the undecane dibasic acid fermentation liquid and removing solid matters; the solids comprise particles of undecanedioic acid.
In the present invention, the salts in the undecanoic acid fermentation treatment liquid include, but are not limited to: one or more of potassium salt, sodium salt, magnesium salt, calcium salt, iron salt, ammonium salt, hydrochloride, carbonate, sulfate, nitrate and phosphate. The salt in the undecanoic acid fermentation treatment liquid includes inorganic salts, and may include a very small amount of soluble salts of undecanoic acid.
In a preferred embodiment of the present invention, the salt in the fermentation treatment solution comprises an inorganic salt. The inorganic salts constitute the vast majority of the total salt. The salt in the fermentation treatment liquid may further include a salt of a dibasic acid. The salts of the dibasic acids account for a very small amount of the total salt.
In a preferred embodiment of the present invention, the salt content in the fermentation treatment solution may be: 8000ppm,7000ppm,6000ppm,5000ppm,4000ppm,3000ppm,2000ppm,1000ppm,800ppm,500ppm.
In a preferred embodiment of the present invention, the salt content in the fermentation treatment liquid is 6000ppm or less.
According to a preferable technical scheme of the invention, the fermentation treatment liquid is a mixed solution obtained by acidifying the fermentation liquid and removing solids. The solids comprise particles of undecanedioic acid; or the solid comprises undecanedioic acid particles and thalli. The fermentation treatment liquid may or may not include bacterial cells. The undecanedioic acid particles comprise a large amount of undecanedioic acid crystals, an amorphous form of undecanedioic acid, and possibly also a very small amount of undecanedioic acid solids in the form of a salt, etc.
The present invention will be described in detail below by way of examples. In the following examples, various raw materials used are commercially available without specific description.
In the following examples, candida Tropicalis (Candida Tropicalis) CAT H1614 was used with a accession number of CCTCC M2015303; or the used Candida Tropicalis (Candida Tropicalis) with the preservation number of CCTCC M203052; the adopted Candida sake CAT H4014 has a preservation number of CCTCC M2011487; the Candida virginiana (Candida viswanathii) used has a preservation number of CCTCC NO: m2020048.
In the following examples, the content of undecanedioic acid in the fermentation broth was determined by GC, specifically, the fermentation broth was subjected to a series of treatments and then separated in a capillary column, and the content of undecanedioic acid in the fermentation broth was determined according to the different peak times of different compounds.
Viable cell concentration was monitored using a hamilton viable cell concentration analyzer.
The method for detecting the salt content in the fermentation liquor of the embodiment of the invention comprises the following steps: and centrifuging or filtering the fermentation liquor to obtain supernatant, placing the supernatant into a glass evaporation dish after drying and constant weight, and evaporating in a water bath. If the residue is colored, dropping hydrogen peroxide until bubbles disappear, evaporating in water bath, and repeating the treatment for several times until the color turns white or is stable. The evaporated evaporation pan is dried to constant weight and weighed. The salt content was calculated.
The detection method of the salt content in the fermentation treatment liquid of the embodiment of the invention comprises the following steps: and (4) placing the fermentation treatment solution in a glass evaporation dish after drying and constant weight, and evaporating in a water bath. If the residue is colored, dropping hydrogen peroxide until bubbles disappear, evaporating in water bath, and repeating the treatment for several times until the color turns white or is stable. And drying the evaporated evaporating dish to constant weight, and weighing. The salt content was calculated.
The technical solution of the present invention is further specifically described below by way of examples.
Examples 1A to 1G
1. Adjusting pH of 10 Baume degree wort to 5.4, weighing 80mL, placing into 500mL conical flask, sterilizing at 121 deg.C for 20min, inoculating 1 seed of Candida tropicalis (Candida tropicalis) CATH 1614 in 2mL glycerine tube stored in-80 deg.C refrigerator, activating and culturing on rotary shaker at 29 deg.C and 250rpm for 42HBody OD 620 The value reached 0.6 (diluted 30 times) or more, and the activation culture was completed.
2. Inoculating the shake flask seeds obtained in the step 1 into a seed tank filled with a seed culture medium, culturing, wherein the inoculation amount is 1.33% (v/v), the temperature in the culture process is controlled to be 29 ℃, the air volume is controlled to be 0.5vvm, and the pressure is controlled to be 0.1MPa, until the OD of the obtained seed liquid is 30 times that of the obtained seed liquid when the seed liquid is diluted 620 The value reaches more than 0.6 (diluted by 30 times).
Wherein, the components of the seed culture medium are as follows: sucrose 2.3%, urea 0.4%, monopotassium phosphate 0.8%, yeast extract 0.5%, corn steep liquor 0.3% (purchased from Kaiser (Jinxiang) biomaterial Co., ltd., total nitrogen content 2.5%, the same below) and polyether defoamer 0.05%.
3. And (3) inoculating the seed solution obtained in the step (2) into a 10L fermentation tank filled with a fermentation culture medium, and performing fermentation tank experiments, wherein the inoculation amount is 10% (v/v), the temperature in the fermentation process is 29 ℃, the air flow control in the fermentation process is 0.18vvm, the pressure is 0.1MPa, and the dissolved oxygen is maintained at about 30% in the fermentation process.
Wherein, the components of the fermentation medium are as follows: 3.0% of glucose, 0.2% of ammonium sulfate, 0.3% of magnesium sulfate heptahydrate, 0.5% of monopotassium phosphate, 0.3% of yeast extract, 0.4% of potassium nitrate, 0.6% of corn steep liquor and 0.05% of polyether type defoamer.
After the seed liquid is inoculated into a fermentation tank, glucose with the concentration of 50% (w/v) is supplemented into the fermentation tank, the supplementing time period is 6-11h, the sugar concentration in the fermentation liquid is respectively controlled in different ranges by controlling the sugar supplementing speed, the thalli begin to grow and propagate rapidly within 13h from the beginning of the fermentation, and when the thalli OD is up 620 When the value reaches more than 0.8 (diluted by 30 times), adjusting the pH value to 6.0, simultaneously adding the undecane hydrocarbon into the fermentation tank in a flowing manner, controlling the flow acceleration rate of the undecane hydrocarbon to maintain the concentration of the undecane hydrocarbon in the fermentation liquor at 1.5% (w/v), maintaining the concentration of living cells at a certain level, stopping adding the undecane hydrocarbon 30h before the end of the fermentation, wherein the adding amount of the undecane hydrocarbon is 1100g, and stopping the fermentation when the alkane is completely exhausted. The fermentation effect of the comparative fermentation broths with different sugar concentrations is shown in table 1.
TABLE 1
As can be seen from Table 1, when the sugar concentration in the fermentation broth is low, the OD value in the fermentation process is low, the viable cell concentration is low, the fermentation period is prolonged, and the final acid yield is low; when the sugar concentration in the fermentation liquor is higher, because the sugar and the substrate alkane exist in the fermentation liquor at the same time, the bacteria use the sugar first and then produce acid, although the influence on the acid production is not large, the fermentation period is prolonged; preferably, when the sugar concentration in the fermentation broth is maintained at about 0.6% (w/v), the acid production is high and the fermentation period is short.
Examples 2A to 2F
The same method as that of example 1 was used except that, in step 3, after the seed solution was inoculated into the fermentation tank, glucose was added to the fermentation tank at a concentration of 50% (w/v) for a period of 6-11 hours, the sugar concentration in the fermentation broth was controlled at a rate of 0.6% (w/v) by controlling the sugar addition rate, and when the bacterial cells OD started to grow and propagate rapidly within 13 hours after the start of fermentation, the bacterial cells OD were 620 When the value reaches more than 0.8 (diluted by 30 times), the pH value is respectively adjusted to 4.5, 5.0, 5.5, 6.0, 6.5 and 7.0, simultaneously, the undecane is fed into the fermentation tank, the feeding speed of the undecane is controlled so as to maintain the concentration of the undecane in the fermentation liquor at 1.5% (w/v), the concentration of the living cells at a certain level, the feeding of the undecane is stopped 30h before the fermentation is finished, the adding amount of the undecane is 1100g, and the fermentation is stopped when the alkane is completely exhausted. The results of the fermentation comparing different pH values are shown in Table 2.
TABLE 2
As can be seen from table 2, with the increase of pH, the viable cell concentration decreases, the final acid yield of fermentation gradually increases, the alkali consumption gradually decreases, and the fermentation period gradually increases. When the pH value exceeds 6.5, the growth of thalli is influenced, the concentration of living cells is obviously reduced, the acid production is reduced, and the fermentation period is prolonged. Preferably, the acid production is higher when the pH value is 5.5-6.5, and the unit consumption of alkali is lower.
Examples 3A to 3E
The same method as that of example 1 was used except that, in step 3, after the seed solution was inoculated into the fermentation tank, glucose was added to the fermentation tank at a concentration of 50% (w/v) for a period of 6-11 hours, the sugar concentration in the fermentation broth was controlled at a rate of 0.6% (w/v) by controlling the sugar addition rate, and when the bacterial cells OD started to grow and propagate rapidly within 13 hours after the start of fermentation, the bacterial cells OD were 620 When the value reaches more than 0.8 (diluted by 30 times), the pH value is adjusted to 6.0, meanwhile, the undecane hydrocarbon is added into the fermentation tank in a flowing mode, the flow adding speed of the undecane hydrocarbon is controlled to enable the concentration of the undecane hydrocarbon in the fermentation liquor to be in different ranges, the concentration of living cells in the fermentation liquor is maintained at a certain level, the undecane hydrocarbon is stopped being supplemented 30 hours before the fermentation is finished, the adding amount of the undecane hydrocarbon is 1100g, and the fermentation is stopped when the alkane is completely exhausted. The fermentation effect of different ranges of the concentration of undecane in the comparative fermentation broths is shown in table 3.
TABLE 3
As can be seen from Table 3, when the concentration of the substrate undecane hydrocarbon is less than 0.5% (w/v), the substrate concentration is too low, which results in slow acid production, decreased acid production and prolonged fermentation period; when the concentration of the substrate undecane hydrocarbon is higher than 3% (w/v), the bacteria can be inhibited from producing acid, and the fermentation period is prolonged; preferably, when the concentration of the substrate, undecane alkane, is in the range of 1.0 to 3.0% (w/v), the effect of the acid production amount and the fermentation period is good.
Examples 4A to 4I
The same method as that of example 1 was used, except that in step 3, after the seed solution was inoculated into the fermentation tank, glucose was added to the fermentation tank at a concentration of 50% (w/v) for a period of 6-11 hours, the sugar concentration in the fermentation broth was controlled at a rate of 0.6% (w/v) by controlling the sugar addition rate, and the bacterial cells began to grow and propagate rapidly from the beginning of the fermentation to 13 hours, when the bacterial cells OD was reached 620 When the value reaches more than 0.8 (diluted by 30 times), the pH value is adjusted to 6.0, meanwhile, the undecane hydrocarbon is added into the fermentation tank in a flowing mode, the different flow adding speeds of the undecane hydrocarbon are controlled, so that the concentration of the undecane hydrocarbon in the fermentation liquor is maintained at 1.5% (w/v), the concentration of living cells is maintained at a certain level, the undecane hydrocarbon is stopped being supplemented 30 hours before the fermentation is finished, the adding amount of the undecane hydrocarbon is 1100g, and the fermentation is stopped when the alkane is completely exhausted. The results of the fermentation comparing different feed rates for undecane are shown in Table 4.
TABLE 4
As can be seen from Table 4, when the substrate flow rate is too low, the substrate is not sufficiently utilized by the cells, and the acid yield is low; when the feeding speed of the substrate is more than 0.08L/h, the substrate concentration is higher, the thalli are inhibited from maintaining growth and producing acid, the number of living cells is reduced, the fermentation period is prolonged, and the alkali consumption is increased; preferably, the fermentation effect is better when the feeding speed of the substrate is maintained within the range of 0.02-0.08L/h.
Example 5
At 200M 3 In a fermentation tank, the fermentation is carried out by using Candida visfata CCTCC NO: m2020048 is the starting strain, with undecane alkane as substrate fermentation to produce undecane two acid method, except the strain difference, its process conditions and example 1E keep the same.
The online viable cell concentration in the fermentation process is 4.5 x 10 8 About cfu/mL, the final fermentation period is 154h, the fermentation liquor is detected, the concentration of the undecanedioic acid in the fermentation liquor is 166.9g/L, and the unit consumption of the alkali is 0.206g/g.
Example 6
At 800M 3 Fermenting in a fermentation tank with Candida sake (Candida sake) CAT H4014 as starting strain and undecane as substrateThe undecanedioic acid process was identical to example 1E except for the strain.
The online viable cell concentration in the fermentation process is 6.2 x 10 8 About cfu/mL, the final fermentation period is 156h, the fermentation liquor is detected, the concentration of the undecanedioic acid in the fermentation liquor is 167.6g/L, and the alkali consumption is 0.192g/g.
Comparative example 1
In the method for producing the undecane diacid by fermenting the candida tropicalis CCTCC M203052 serving as a starting strain and the undecane alkane serving as a substrate in a 10L fermentation tank, the process conditions are kept consistent with those in example 1E except that the pH is controlled to be different in the fermentation process.
During the fermentation conversion process by adding the undecane, the pH is controlled at 8.4, and the online viable cell concentration is lower at 5.4 x 10 during the fermentation process 7 Around cfu/mL, fermentation was stopped when the alkane was completely depleted. And the final fermentation period is 188h, and the fermentation liquor is detected, wherein the concentration of the undecane diacid in the fermentation liquor is 156.6g/L, and the alkali consumption is 1.215g/g.
Taking the fermentation liquid of the example 4C and the fermentation liquid of the comparative example 1, and removing thalli and undecanedioic acid by acidification and crystallization according to a conventional method in the field to obtain a fermentation treatment liquid. The results of measuring the salt content in the fermentation broth and the fermentation treatment liquid are shown in Table 5. The step effectively removes thalli, residual hydrocarbon and metabolic impurities which influence downstream polymerization, such as undecanoic monoacid and the like in the fermentation liquor.
TABLE 5
| Examples of the invention | Content of salts in fermentation broth/%) | Salt content/ppm in fermentation treatment liquid |
| Example 4C | 7.83 | 5850 |
| Comparative example 1 | ＞100 | 88670 |
As can be seen from Table 5, compared with the conventional process in comparative example 1, the method provided by the present invention can significantly reduce the salt content in the obtained fermentation broth and fermentation treatment solution, and can significantly reduce the subsequent purification process, thereby reducing the production cost and environmental stress.
Example 7
The preparation method of the undecane diacid comprises the following steps:
adjusting the pH of the fermentation broth obtained in example 4C to 3.5 with sulfuric acid to obtain a solid substance and a fermentation treatment solution, performing centrifugal separation to obtain the solid substance, dissolving the solid substance in acetic acid, and controlling the ratio of the solid substance to the acetic acid to be 1:3, adding activated carbon which accounts for about 1 percent of the volume of the clear liquid, decoloring for 80min at 95 ℃, and filtering and separating to obtain undecanedioic acid clear liquid; cooling the clear liquid to 70 ℃, preserving the heat for 1.5 hours, then cooling to 30 ℃, and crystallizing; and centrifugally separating to obtain the undecane diacid product with the purity of more than 98 percent.
The preferred embodiments of the present invention have been described above in detail, but the present invention is not limited thereto. Within the scope of the technical idea of the invention, many simple modifications can be made to the technical solution of the invention, including combinations of various technical features in any other suitable way, and these simple modifications and combinations should also be regarded as the disclosure of the invention, and all fall within the scope of the invention.
Claims (17)
1. A preparation method of undecanedioic acid fermentation broth is characterized by comprising the following steps: performing seed culture and fermentation culture on yeast, adding undecane hydrocarbon during the fermentation culture process, and controlling pH of the fermentation liquid to be below 7.0, further 4.5-7.0 to obtain undecane diacid fermentation liquid.
2. The method according to claim 1, characterized in that the pH of the fermentation broth is controlled to a pH value of 5.0 to 7.0, preferably 5.5 to 6.5.
3. The method according to claim 1 or 2, wherein the OD of the bacterial cells is 620 When the value reaches more than 0.6-1.0 (diluted by 30 times), adding the undecane to start fermentation and conversion; and/or
In the fermentation culture process, the temperature is controlled to be 26-35 ℃, and preferably 28-32 ℃; the air volume is 0.1-1.0vvm, preferably 0.1-0.6vvm; the pressure is 0.05-0.15MPa, preferably 0.08-0.12MPa; and/or
In the fermentation culture process, the dissolved oxygen is controlled to be more than 15 percent, preferably 20 to 40 percent; and/or
During the fermentation culture, the concentration of the undecane in the fermentation liquid is controlled to be 0.1-5% (w/v), preferably 1-3% (w/v).
4. The method according to any of claims 1 to 3, wherein the concentration of viable cells in the fermentation broth is controlled to 3 x 10 during the fermentation culture 7 cfu/mL-4*10 9 cfu/mL, preferably 2 x 10 8 cfu/mL-8*10 8 cfu/mL。
5. The method according to any one of claims 1 to 4, wherein when the seed culture and the fermentation culture are carried out, a seed tank culture medium and a fermenter culture medium comprising a carbon source, a nitrogen source, an inorganic salt and an antifoaming agent are used; wherein,
the carbon source includes but is not limited to one or more of glucose, sucrose and maltose; and/or the presence of a gas in the gas,
the nitrogen source comprises but is not limited to one or more of yeast extract, ammonium sulfate, urea, corn steep liquor, potassium nitrate and yeast powder; and/or
The inorganic salt comprises but is not limited to one or more of magnesium sulfate heptahydrate, potassium nitrate, ferrous sulfate, sodium chloride, dipotassium hydrogen phosphate, magnesium sulfate, sodium dihydrogen phosphate, sodium chloride and potassium dihydrogen phosphate; and/or
The defoaming agent includes, but is not limited to, a silicone type defoaming agent and/or a polyether type defoaming agent.
6. The method according to any one of claims 1 to 5, wherein the temperature is controlled during seed cultivation to be 26-32 ℃, preferably 28-30 ℃; the air volume is 0.1-0.8vvm, preferably 0.4-0.6vvm; the pressure is 0.05-0.15MPa, preferably 0.08-0.12MPa; and/or
When the seed liquid OD 620 When the value is more than 0.6 (diluted by 30 times), inoculating the seed liquid into a fermentation culture medium for fermentation culture, wherein the inoculation amount of the seed liquid is 5-35% (v/v), preferably 10-30% (v/v).
7. The method of any one of claims 1 to 6, wherein said yeast comprises Candida Tropicalis (Candida Tropicalis), candida sake (Candida sake) or Candida virginiana (Candida viswanathii).
8. The method according to any one of claims 1-7, further comprising: in the fermentation culture process, a primary carbon source is supplemented to the fermentation liquor;
further, the sugar concentration in the fermentation broth is controlled to be 0.01-5% (w/v), preferably 0.1-0.8% (w/v), and more preferably 0.5-0.8% (w/v), by supplementing the primary carbon source; and/or
The concentration of the primary carbon source is 20-70% (w/v), preferably 40-50% (w/v); and/or
The time period for supplementing the primary carbon source to the fermentation liquor is 5-14h, preferably 6-11h; and/or
The primary carbon source is preferably glucose.
9. A method for preparing undecanedioic acid, comprising: the method of any one of claims 1 to 8, wherein the undecanedioic acid fermentation broth is obtained and then subjected to extraction and purification, or the undecanedioic acid fermentation broth is treated to obtain a fermentation treatment solution, and then subjected to extraction and purification; to obtain the undecane diacid.
10. The method of claim 9, wherein the method of extraction and purification comprises: a decoloring step;
preferably, the decolorization step comprises decolorization by using activated carbon, and the added amount of the activated carbon is not more than 5% of the volume of the clear liquid;
the decolorizing temperature is 80-105 deg.C, and the time is 10-170min.
11. The method of claim 9, wherein the method of extraction and purification comprises: forming solid matters and separating the solid matters; preferably, the method for forming the solid is acidification crystallization; the separation steps of forming solids and solids may further include any other extraction and purification steps.
12. The method of claim 10, wherein the decolorizing step comprises: decoloring fermentation liquor or fermentation treatment liquor, or decoloring solution formed by dissolving the solid by an organic solvent;
the organic solvent is selected from one or more of alcohol, acid, ketone and ester, wherein the alcohol is selected from one or more of methanol, ethanol, isopropanol and n-butanol; the acid is acetic acid; the ketone is acetone; the ester is ethyl acetate and/or butyl acetate.
13. The method of claim 9, wherein the method of extraction and purification comprises: forming a solid, dissolving the solid by an organic solvent, separating to obtain a clear liquid, and finally crystallizing and separating the clear liquid to obtain the undecanedioic acid;
preferably, the organic solvent is selected from one or more of alcohols, acids, ketones and esters, wherein the alcohol is selected from one or more of methanol, ethanol, isopropanol and n-butanol; the acid is acetic acid; the ketone is acetone; the ester is ethyl acetate and/or butyl acetate; and/or
The crystallization is cooling crystallization, and the cooling crystallization comprises the following steps: cooling to 60-85 deg.C, maintaining for 1-2 hr, cooling to 20-40 deg.C, and crystallizing.
14. An undecanedioic acid prepared by the method of any one of claims 9-13.
15. An undecabanedioic acid fermentation broth characterized in that the salt content of the undecabanedioic acid fermentation broth is below 15%, preferably below 10%, more preferably below 8%; the percentage is the mass percentage of the total amount of the undecanedioic acid generated by fermentation.
16. An undecandioic acid fermentation broth according to claim 15, characterized in that it is fermentatively transformed at a pH of 7.0 or less, preferably 4.5-7.0, further preferably 5.0-7.0, more preferably 5.5-6.5.
17. The undecanedioic acid fermentation treatment liquid is characterized in that the salt content in the fermentation treatment liquid is 8000ppm or less, preferably 6000ppm or less, wherein the parts per million is the mass parts per million of the salt in the undecanedioic acid fermentation treatment liquid; the fermentation treatment liquid is obtained by acidifying the undecane dibasic acid fermentation liquid according to claim 15 or 16 and removing solids; the solids comprise particles of undecanedioic acid.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202111172639.4A CN115960973A (en) | 2021-10-08 | 2021-10-08 | Undecanedioic acid fermentation liquid, undecanedioic acid fermentation treatment liquid, undecanedioic acid and preparation method thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202111172639.4A CN115960973A (en) | 2021-10-08 | 2021-10-08 | Undecanedioic acid fermentation liquid, undecanedioic acid fermentation treatment liquid, undecanedioic acid and preparation method thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN115960973A true CN115960973A (en) | 2023-04-14 |
Family
ID=87357315
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202111172639.4A Pending CN115960973A (en) | 2021-10-08 | 2021-10-08 | Undecanedioic acid fermentation liquid, undecanedioic acid fermentation treatment liquid, undecanedioic acid and preparation method thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN115960973A (en) |
-
2021
- 2021-10-08 CN CN202111172639.4A patent/CN115960973A/en active Pending
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| US11802298B2 (en) | Decanedioic acid produced by microbial fermentation process and preparation method thereof | |
| Anastassiadis et al. | Gluconic acid production | |
| US11319561B2 (en) | Method for producing a long chain dicarboxylic acid by fermentation, fermentation broth, treated fermentation broth and wastewater | |
| CN110272341B (en) | Purification method of long-chain dibasic acid | |
| CN117210343A (en) | Candida tropicalis and application thereof, and production method of long-chain dibasic acid | |
| CN111748480B (en) | Candida virginiana and application thereof | |
| CN105603018B (en) | Method for indirectly preparing D-sodium erythorbate | |
| Milsom | Organic acids by fermentation, especially citric acid | |
| CN112725385B (en) | Method for preparing long-chain dicarboxylic acid by fermentation | |
| US5686277A (en) | Fermentation process for preparing xylitol using mutant cells | |
| CN107177508B (en) | Method for synthesizing and producing long-chain dodecanedioic acid by biological method | |
| CN115960973A (en) | Undecanedioic acid fermentation liquid, undecanedioic acid fermentation treatment liquid, undecanedioic acid and preparation method thereof | |
| CN111378696A (en) | Fermentation substrate and method for producing long-chain dicarboxylic acid by fermentation of fermentation substrate | |
| CN105713932B (en) | A kind of method that microbial fermentation prepares 2,3- butanediol | |
| CN115595337A (en) | Method for producing high-purity long-chain dicarboxylic acid by using coal-to-paraffin | |
| CN114249645A (en) | Extraction method of sebacic acid and sebacic acid product | |
| CN110683947A (en) | Long-chain dibasic acid with low content of hydroxy acid impurities and method for reducing content of hydroxy acid impurities | |
| CN111850060A (en) | Method for producing hexadecanedioic acid by fermentation, hexadecanedioic acid and preparation method thereof | |
| CN110564785A (en) | method for producing undecanedioic acid by fermentation of candida virescens | |
| CN1353193A (en) | Enzyme method for preparing dextral glycoanhydride | |
| Masheti | Influence Of Oxygen Dissolved On The Biomass Grouth During Fermentation Process | |
| EP3822356A1 (en) | Method for the production of a dicarboxylic acid | |
| CN111718919A (en) | Culture medium for producing esterifying enzyme and preparation method of crude esterifying enzyme preparation | |
| JPH09131199A (en) | Production of extracellular polysaccharide produced with microorganism of genus lipomyces | |
| CN117587087A (en) | Preparation method of liquid natamycin |
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 |