CN103243032A - Sake candida and fermentation method thereof - Google Patents
Sake candida and fermentation method thereof Download PDFInfo
- Publication number
- CN103243032A CN103243032A CN2012100302784A CN201210030278A CN103243032A CN 103243032 A CN103243032 A CN 103243032A CN 2012100302784 A CN2012100302784 A CN 2012100302784A CN 201210030278 A CN201210030278 A CN 201210030278A CN 103243032 A CN103243032 A CN 103243032A
- Authority
- CN
- China
- Prior art keywords
- acid
- candida
- candida sake
- substratum
- sake
- 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
- 238000000034 method Methods 0.000 title claims description 35
- 238000000855 fermentation Methods 0.000 title abstract description 19
- 230000004151 fermentation Effects 0.000 title abstract description 19
- 241000222120 Candida <Saccharomycetales> Species 0.000 title abstract description 5
- 150000007520 diprotic acids Chemical class 0.000 claims description 31
- 241001530515 Candida sake Species 0.000 claims description 29
- 239000002253 acid Substances 0.000 claims description 25
- 150000001335 aliphatic alkanes Chemical class 0.000 claims description 14
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 claims description 12
- 229910052799 carbon Inorganic materials 0.000 claims description 10
- -1 n-dodecane hydrocarbon Chemical class 0.000 claims description 10
- QJGQUHMNIGDVPM-UHFFFAOYSA-N nitrogen group Chemical group [N] QJGQUHMNIGDVPM-UHFFFAOYSA-N 0.000 claims description 10
- 239000004215 Carbon black (E152) Substances 0.000 claims description 9
- 229930195733 hydrocarbon Natural products 0.000 claims description 9
- XSQUKJJJFZCRTK-UHFFFAOYSA-N Urea Chemical compound NC(N)=O XSQUKJJJFZCRTK-UHFFFAOYSA-N 0.000 claims description 8
- 239000004202 carbamide Substances 0.000 claims description 8
- 239000000758 substrate Substances 0.000 claims description 8
- 229940041514 candida albicans extract Drugs 0.000 claims description 7
- 239000012138 yeast extract Substances 0.000 claims description 7
- 229940094933 n-dodecane Drugs 0.000 claims description 6
- QGZKDVFQNNGYKY-UHFFFAOYSA-N Ammonia Chemical compound N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 claims description 4
- 239000001888 Peptone Substances 0.000 claims description 4
- 108010080698 Peptones Proteins 0.000 claims description 4
- 240000008042 Zea mays Species 0.000 claims description 4
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 claims description 4
- 235000002017 Zea mays subsp mays Nutrition 0.000 claims description 4
- 125000004432 carbon atom Chemical group C* 0.000 claims description 4
- 235000005822 corn Nutrition 0.000 claims description 4
- 235000019319 peptone Nutrition 0.000 claims description 4
- 125000001477 organic nitrogen group Chemical group 0.000 claims description 3
- 229910002651 NO3 Inorganic materials 0.000 claims description 2
- NHNBFGGVMKEFGY-UHFFFAOYSA-N Nitrate Chemical compound [O-][N+]([O-])=O NHNBFGGVMKEFGY-UHFFFAOYSA-N 0.000 claims description 2
- IOVCWXUNBOPUCH-UHFFFAOYSA-M Nitrite anion Chemical compound [O-]N=O IOVCWXUNBOPUCH-UHFFFAOYSA-M 0.000 claims description 2
- 108010009736 Protein Hydrolysates Proteins 0.000 claims description 2
- 235000019764 Soybean Meal Nutrition 0.000 claims description 2
- 150000001413 amino acids Chemical class 0.000 claims description 2
- 229910021529 ammonia Inorganic materials 0.000 claims description 2
- 235000011114 ammonium hydroxide Nutrition 0.000 claims description 2
- 150000003863 ammonium salts Chemical class 0.000 claims description 2
- 108090000765 processed proteins & peptides Proteins 0.000 claims description 2
- 239000004455 soybean meal Substances 0.000 claims description 2
- 230000001580 bacterial effect Effects 0.000 abstract description 20
- 238000004519 manufacturing process Methods 0.000 abstract description 4
- 108090000623 proteins and genes Proteins 0.000 description 14
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 11
- 239000002609 medium Substances 0.000 description 11
- 239000000126 substance Substances 0.000 description 11
- 238000010586 diagram Methods 0.000 description 10
- 239000000203 mixture Substances 0.000 description 10
- 239000000243 solution Substances 0.000 description 9
- 238000011218 seed culture Methods 0.000 description 8
- 238000005303 weighing Methods 0.000 description 8
- 241000894006 Bacteria Species 0.000 description 7
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 7
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 6
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 6
- 239000003513 alkali Substances 0.000 description 6
- 238000005516 engineering process Methods 0.000 description 6
- 239000001963 growth medium Substances 0.000 description 6
- 150000002632 lipids Chemical class 0.000 description 6
- 239000000463 material Substances 0.000 description 6
- 230000001954 sterilising effect Effects 0.000 description 6
- 238000004659 sterilization and disinfection Methods 0.000 description 6
- 238000012360 testing method Methods 0.000 description 6
- 241000222178 Candida tropicalis Species 0.000 description 5
- 108091028043 Nucleic acid sequence Proteins 0.000 description 5
- 239000007788 liquid Substances 0.000 description 5
- 239000000047 product Substances 0.000 description 5
- 102000004169 proteins and genes Human genes 0.000 description 5
- 101150053659 POX4 gene Proteins 0.000 description 4
- 101150004239 POX5 gene Proteins 0.000 description 4
- 230000008901 benefit Effects 0.000 description 4
- 239000011248 coating agent Substances 0.000 description 4
- 238000000576 coating method Methods 0.000 description 4
- YCOZIPAWZNQLMR-UHFFFAOYSA-N pentadecane Chemical compound CCCCCCCCCCCCCCC YCOZIPAWZNQLMR-UHFFFAOYSA-N 0.000 description 4
- 239000002504 physiological saline solution Substances 0.000 description 4
- 238000003822 preparative gas chromatography Methods 0.000 description 4
- 230000008569 process Effects 0.000 description 4
- 239000007858 starting material Substances 0.000 description 4
- 239000006228 supernatant Substances 0.000 description 4
- WGTYBPLFGIVFAS-UHFFFAOYSA-M tetramethylammonium hydroxide Chemical compound [OH-].C[N+](C)(C)C WGTYBPLFGIVFAS-UHFFFAOYSA-M 0.000 description 4
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 4
- FRXSZNDVFUDTIR-UHFFFAOYSA-N 6-methoxy-1,2,3,4-tetrahydroquinoline Chemical compound N1CCCC2=CC(OC)=CC=C21 FRXSZNDVFUDTIR-UHFFFAOYSA-N 0.000 description 3
- 229920001817 Agar Polymers 0.000 description 3
- 229930006000 Sucrose Natural products 0.000 description 3
- 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 3
- 239000008272 agar Substances 0.000 description 3
- 239000007864 aqueous solution Substances 0.000 description 3
- 238000010790 dilution Methods 0.000 description 3
- 239000012895 dilution Substances 0.000 description 3
- 235000011187 glycerol Nutrition 0.000 description 3
- 239000003921 oil Substances 0.000 description 3
- 238000007254 oxidation reaction Methods 0.000 description 3
- 238000012856 packing Methods 0.000 description 3
- 238000002360 preparation method Methods 0.000 description 3
- 238000012216 screening Methods 0.000 description 3
- 239000007787 solid Substances 0.000 description 3
- 239000005720 sucrose Substances 0.000 description 3
- BGHCVCJVXZWKCC-UHFFFAOYSA-N tetradecane Chemical compound CCCCCCCCCCCCCC BGHCVCJVXZWKCC-UHFFFAOYSA-N 0.000 description 3
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- 101150033406 CYP52A12 gene Proteins 0.000 description 2
- 101100161762 Candida tropicalis POX5 gene Proteins 0.000 description 2
- 101100168396 Debaryomyces hansenii CYP52A12 gene Proteins 0.000 description 2
- 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 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
- 238000010170 biological method Methods 0.000 description 2
- 239000006227 byproduct Substances 0.000 description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- 238000001816 cooling Methods 0.000 description 2
- TVIDDXQYHWJXFK-UHFFFAOYSA-N dodecanedioic acid Chemical compound OC(=O)CCCCCCCCCCC(O)=O TVIDDXQYHWJXFK-UHFFFAOYSA-N 0.000 description 2
- 238000011156 evaluation Methods 0.000 description 2
- 238000000605 extraction Methods 0.000 description 2
- 238000012262 fermentative production Methods 0.000 description 2
- 239000012467 final product Substances 0.000 description 2
- 239000000499 gel Substances 0.000 description 2
- 239000008103 glucose Substances 0.000 description 2
- 238000011081 inoculation Methods 0.000 description 2
- 239000002655 kraft paper Substances 0.000 description 2
- KWGKDLIKAYFUFQ-UHFFFAOYSA-M lithium chloride Chemical compound [Li+].[Cl-] KWGKDLIKAYFUFQ-UHFFFAOYSA-M 0.000 description 2
- 239000000314 lubricant Substances 0.000 description 2
- 230000035800 maturation Effects 0.000 description 2
- 230000004060 metabolic process Effects 0.000 description 2
- 239000000178 monomer Substances 0.000 description 2
- 150000007518 monoprotic acids Chemical class 0.000 description 2
- 239000012071 phase Substances 0.000 description 2
- FGIUAXJPYTZDNR-UHFFFAOYSA-N potassium nitrate Chemical compound [K+].[O-][N+]([O-])=O FGIUAXJPYTZDNR-UHFFFAOYSA-N 0.000 description 2
- 239000000843 powder Substances 0.000 description 2
- 238000004321 preservation Methods 0.000 description 2
- 239000002994 raw material Substances 0.000 description 2
- 239000002002 slurry Substances 0.000 description 2
- 239000002689 soil Substances 0.000 description 2
- 238000003860 storage Methods 0.000 description 2
- 238000003786 synthesis reaction Methods 0.000 description 2
- 239000008399 tap water Substances 0.000 description 2
- 235000020679 tap water Nutrition 0.000 description 2
- 230000009466 transformation Effects 0.000 description 2
- 238000005406 washing Methods 0.000 description 2
- QFGCFKJIPBRJGM-UHFFFAOYSA-N 12-[(2-methylpropan-2-yl)oxy]-12-oxododecanoic acid Chemical compound CC(C)(C)OC(=O)CCCCCCCCCCC(O)=O QFGCFKJIPBRJGM-UHFFFAOYSA-N 0.000 description 1
- HCUZVMHXDRSBKX-UHFFFAOYSA-N 2-decylpropanedioic acid Chemical compound CCCCCCCCCCC(C(O)=O)C(O)=O HCUZVMHXDRSBKX-UHFFFAOYSA-N 0.000 description 1
- NIXOWILDQLNWCW-UHFFFAOYSA-N Acrylic acid Chemical compound OC(=O)C=C NIXOWILDQLNWCW-UHFFFAOYSA-N 0.000 description 1
- 101100433727 Caenorhabditis elegans got-1.2 gene Proteins 0.000 description 1
- 108090000201 Carboxypeptidase B2 Proteins 0.000 description 1
- 108020004414 DNA Proteins 0.000 description 1
- 101100168397 Debaryomyces hansenii CYP52A13 gene Proteins 0.000 description 1
- 239000004831 Hot glue Substances 0.000 description 1
- OVRNDRQMDRJTHS-UHFFFAOYSA-N N-acelyl-D-glucosamine Natural products CC(=O)NC1C(O)OC(CO)C(O)C1O OVRNDRQMDRJTHS-UHFFFAOYSA-N 0.000 description 1
- OVRNDRQMDRJTHS-FMDGEEDCSA-N N-acetyl-beta-D-glucosamine Chemical compound CC(=O)N[C@H]1[C@H](O)O[C@H](CO)[C@@H](O)[C@@H]1O OVRNDRQMDRJTHS-FMDGEEDCSA-N 0.000 description 1
- MBLBDJOUHNCFQT-LXGUWJNJSA-N N-acetylglucosamine Natural products CC(=O)N[C@@H](C=O)[C@@H](O)[C@H](O)[C@H](O)CO MBLBDJOUHNCFQT-LXGUWJNJSA-N 0.000 description 1
- 239000004677 Nylon Substances 0.000 description 1
- 239000004952 Polyamide Substances 0.000 description 1
- DPDMMXDBJGCCQC-UHFFFAOYSA-N [Na].[Cl] Chemical compound [Na].[Cl] DPDMMXDBJGCCQC-UHFFFAOYSA-N 0.000 description 1
- 238000002479 acid--base titration Methods 0.000 description 1
- 150000007513 acids Chemical class 0.000 description 1
- 230000004913 activation Effects 0.000 description 1
- DCAYPVUWAIABOU-UHFFFAOYSA-N alpha-n-hexadecene Natural products CCCCCCCCCCCCCCCC DCAYPVUWAIABOU-UHFFFAOYSA-N 0.000 description 1
- 238000013459 approach Methods 0.000 description 1
- 239000002585 base Substances 0.000 description 1
- 230000003851 biochemical process Effects 0.000 description 1
- 150000001721 carbon Chemical group 0.000 description 1
- 239000003054 catalyst Substances 0.000 description 1
- 235000019219 chocolate Nutrition 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 239000010779 crude oil Substances 0.000 description 1
- 239000003085 diluting agent Substances 0.000 description 1
- SNRUBQQJIBEYMU-UHFFFAOYSA-N dodecane Chemical compound CCCCCCCCCCCC SNRUBQQJIBEYMU-UHFFFAOYSA-N 0.000 description 1
- 238000005265 energy consumption Methods 0.000 description 1
- 229920006351 engineering plastic Polymers 0.000 description 1
- 150000002148 esters Chemical class 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 239000000284 extract Substances 0.000 description 1
- 238000004108 freeze drying Methods 0.000 description 1
- 238000012268 genome sequencing Methods 0.000 description 1
- QQHJDPROMQRDLA-UHFFFAOYSA-N hexadecanedioic acid Chemical compound OC(=O)CCCCCCCCCCCCCCC(O)=O QQHJDPROMQRDLA-UHFFFAOYSA-N 0.000 description 1
- IIYFAKIEWZDVMP-UHFFFAOYSA-N linear paraffin C13 Natural products CCCCCCCCCCCCC IIYFAKIEWZDVMP-UHFFFAOYSA-N 0.000 description 1
- 238000003760 magnetic stirring Methods 0.000 description 1
- 230000014759 maintenance of location Effects 0.000 description 1
- 235000013372 meat Nutrition 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 230000000813 microbial effect Effects 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 238000012544 monitoring process Methods 0.000 description 1
- 125000000896 monocarboxylic acid group Chemical group 0.000 description 1
- 231100000350 mutagenesis Toxicity 0.000 description 1
- 238000002703 mutagenesis Methods 0.000 description 1
- 231100000219 mutagenic Toxicity 0.000 description 1
- 230000003505 mutagenic effect Effects 0.000 description 1
- 229950006780 n-acetylglucosamine Drugs 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- 238000010606 normalization Methods 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 235000016709 nutrition Nutrition 0.000 description 1
- 230000035764 nutrition Effects 0.000 description 1
- 229920001778 nylon Polymers 0.000 description 1
- 238000005457 optimization Methods 0.000 description 1
- 230000003647 oxidation Effects 0.000 description 1
- 230000037361 pathway Effects 0.000 description 1
- 239000002304 perfume Substances 0.000 description 1
- 239000004014 plasticizer Substances 0.000 description 1
- 229920002647 polyamide Polymers 0.000 description 1
- 239000004417 polycarbonate Substances 0.000 description 1
- 229920000515 polycarbonate Polymers 0.000 description 1
- 235000010482 polyoxyethylene sorbitan monooleate Nutrition 0.000 description 1
- 229920000053 polysorbate 80 Polymers 0.000 description 1
- 235000010333 potassium nitrate Nutrition 0.000 description 1
- 239000004323 potassium nitrate Substances 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 238000012163 sequencing technique Methods 0.000 description 1
- 239000009671 shengli Substances 0.000 description 1
- 235000014347 soups Nutrition 0.000 description 1
- 235000013599 spices Nutrition 0.000 description 1
- 230000007480 spreading Effects 0.000 description 1
- 238000003892 spreading Methods 0.000 description 1
- 230000000087 stabilizing effect Effects 0.000 description 1
- 238000003756 stirring Methods 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
- 238000001308 synthesis method Methods 0.000 description 1
Images
Abstract
The invention relates to a novel sake candida bacterial strain and a method for production of long chain diacid by fermentation of the novel sake candida bacterial strain.
Description
Invention field
The present invention relates to the method for candida sake (Candida sake) and fermentative production long-chain biatomic acid thereof.
Background technology
Long-chain biatomic acid (LCDA is also referred to as long chain dicarboxylic acid and long chain diacid) comprises chemical formula HOOC (CH2)
nThe diprotic acid of COOH, wherein n 〉=7.LCDA is the base monomer raw material of a series of extraordinary synthetic materialss.Long-chain biatomic acid and derivative monomer thereof can be the important source material of synthetic perfume, engineering plastics, cold resistant plasticizer, senior lubricant and products such as polyamide hot, powder coating for the production of extraordinary nylon, polycarbonate, powder coating, spices, hot melt adhesive, extraordinary lubricant etc.
There is the chemical synthesis process of multiple long-chain biatomic acid in this area, is the mixture of long-chain biatomic acid and short chain diprotic acid but using method is not easy and major part obtains.Need be further purified step when therefore, producing long-chain biatomic acid with these methods.Well known microbial transformation alkane, lipid acid or its ester of utilizing produced long-chain biatomic acid.Because the restriction of existing biological method, chemosynthesis remain the optimization approach of producing long-chain biatomic acid.
This area is general uses several yeast strains to carry out the fermentative production long-chain biatomic acid, and when using alkane or lipid acid to cultivate as carbon source, long-chain biatomic acid produces as by product.Yeast has three Biochemical processes when alkane and lipid acid are carried out metabolism: alkane is to the alpha-oxidation of lipid acid, and lipid acid is to alpha-, and the omega oxidation of omega-dicarboxylic acid and lipid acid are to CO
2Degraded type beta-oxidation with water.With respect to abiotic method for transformation, the bioconversion method of producing diprotic acid has multiple potential advantages.Wherein mainly be to use reproducible raw material as parent material and produce the ability that does not produce harmful chemical by-product in the diprotic acid process.Use another considerable advantage of biological method to be that this method can easily regulate to utilize identical biological catalyst and identical device to produce multiple diprotic acid.Because existing organic chemical synthesis only is suitable for manufacture order kind diprotic acid, the synthetic exploitation that will need the new synthetic schemes of each diprotic acid of several different diprotic acid.On the other hand, the yeast bio catalyzer can utilize identical device, substratum and step to produce the diprotic acid of all lengths, and difference only is to provide the substrate of different carbon atom length to yeast.
Though can utilize various technology to improve for example output of candida tropicalis (Candida tropicalis) fermentation diprotic acid of existing yeast, but can also comprise dodecanedioic acid, undecane dicarboxylic acid, tetradecane diacid, pentadecane diacid and Thapsic acid by providing new candida bacterial strain to produce the long-chain biatomic acid of higher output yield.Therefore an object of the present invention is to provide new bacterial strain and utilize these bacterial strains to produce the method for one or more long-chain biatomic acids.Use this candiyeast to produce long-chain biatomic acid substituted chemistry synthesis method fully, and have the output height, less energy-consumption, the advantage that saves material.
Summary of the invention
In one embodiment of the invention, provide candida sake CAT H4012CCTCC M2011485.Described bacterial classification is preserved in Chinese typical culture collection center (Chinese Wuhan Wuhan University) on December 29th, 2011, and deposit number is: CCTCC M 2011485.
In another embodiment of the present invention, the method of utilizing described candida sake CATH4012CCTCC M 2011485 to produce diprotic acid is provided, and said method comprising the steps of: a. cultivates candida sake CATH4012 in the substratum that contains at least a nitrogenous source and at least a organic substrates; Reclaim the diprotic acid of cultivating the step from a with b..
In yet another embodiment of the invention, the method of utilizing described candida sake CATH4012CCTCC M 2011485 to produce diprotic acid is provided, said method comprising the steps of: a. cultivates candida sake CATH4012 in the substratum that contains at least a nitrogenous source and at least a organic substrates, and wherein said at least a organic substrates is selected from alkane with 12 to 16 carbon atoms or the normal alkane of 12 to 16 carbon atoms; Reclaim the diprotic acid of cultivating the step from a with b..
In the above-described embodiment, described diprotic acid at least a or positive 12 carbon that are 12 carbon to the 16-dicarboxylic acid are at least a to the 16-dicarboxylic acid.
In yet another embodiment of the invention, the method of utilizing described candida sake CATH4012CCTCC M 2011485 to produce diprotic acid is provided, and said method comprising the steps of: a. cultivates candida sake CAT H4012 in the substratum that contains at least a nitrogenous source and n-dodecane hydrocarbon; Reclaim the n-dodecane diacid of cultivating the step from a with b..
In the above-described embodiment, those skilled in the art know operable nitrogenous source.Can use various organonitrogens and inorganic nitrogen.Organic nitrogen source includes but not limited to yeast extract paste, corn steep liquor, extractum carnis, soybean meal hydrolysate, peptone, urea, each seed amino acid and peptide etc., and preferred nitrogenous source is urea.Inorganic nitrogen-sourced nitrate, ammoniacal liquor, liquefied ammonia, ammonium salt, the nitrite etc. of including but not limited to.
The purposes of candida sake H4012CCTCC M 2011485, it is mainly for the production of the application of long-chain biatomic acid.
Description of drawings
Figure 1 shows that the protein sequence comparison diagram of the POX5 gene of CAT H4012.
Figure 2 shows that the POX5 gene DNA sequence comparison diagram of CAT H4012.
Figure 3 shows that the protein sequence comparison diagram of the POX4 gene of CAT H4012.
Figure 4 shows that the POX4 gene DNA sequence comparison diagram of CAT H4012.
Figure 5 shows that the protein sequence comparison diagram of the CYTb5 gene of CAT H4012.
Figure 6 shows that the CYTb5 gene DNA sequence comparison diagram of CAT H4012.
Figure 7 shows that the protein sequence comparison diagram of the CYP52D2 gene of CAT H4012.
Figure 8 shows that the CYP52D2 gene DNA sequence comparison diagram of CAT H4012.
Figure 9 shows that the protein sequence comparison diagram of the CYP52A12 gene of CAT H4012.
Figure 10 shows that the CYP52A12 gene DNA sequence comparison diagram of CAT H4012.
Specific embodiments
By reading following detailed description, those skilled in the art will be more readily understood these and other embodiment of the present invention, key element and advantage.Should be understood that provides following examples with proof and further explains preferred embodiment more of the present invention and aspect, should not be interpreted as limiting its scope.
Although describe similar or suitable method to this paper and material can be used for practice of the present disclosure or test, this paper has described suitable method and material.Unless otherwise defined, all technology used herein are identical with the implication of disclosure one of ordinary skill in the art common sense with scientific terminology.If conflict is arranged, be as the criterion with the definition of this explanation.
When quantity, concentration or other value or parameter provide with scope or height value list, be interpreted as specific arbitrary all scopes that any high low range boundary is formed that disclose, no matter whether scope discloses separately.When this paper enumerates the scope of numerical value, unless otherwise indicated, this scope is intended to comprise its end points, and all integers and mark in this scope.When the range of definition, unintentionally scope of the present invention is limited to the particular value of enumerating.
When being used for this paper, term " comprises ", " comprising ", " having " or its any other variant, is intended to relate to non-exclusive comprising.For example, comprise technology, method, object or the device of a series of key elements and unnecessarily be limited to only those key elements, but can comprise do not list especially or this type of technology, method, object or install other intrinsic key element.
When " one " or " a kind of " be used for to describe multiple key element and composition herein only for convenient and general sense of the present disclosure is provided.This description is understood to include one or at least one and odd number also comprises plural number, unless obviously it is intended to other.
The material of this paper, method and example be usefulness for illustrative purposes only, unless stated otherwise, is not intended to as restriction.
Embodiment
This paper is, and the part term definition is as follows:
Activation medium
Employing is called a kind of aqueous solution substratum of " YPD substratum ", and it comprises following composition: 20g/L glucose, 10g/L yeast extract and 20g/L peptone.Described " YPD substratum " contains the agar of substratum weight 2%, makes substratum gel at room temperature.Described YPD substratum prepares pH regulator to 7.0-7.5 by water and 1N sodium hydroxide solution.With this substratum sterilization 20min under 121 ℃.
The shake-flask seed substratum
Another substratum that is suitable for cultivating bacterial strain of the present invention is " seed culture medium ", and it is a kind of aqueous solution substratum, comprises following composition: 10-30g/L sucrose, 1.5-10g/L corn slurries, 1-10g/L yeast extract, 4-12g/L KH
2PO
4, the heavy wax of 0.5-5g/L urea and 0-50ml/L.Described substratum water preparation, and at 121 ℃ of 20min that sterilize down.Urea is sterilized separately, sterilizes 15 minutes down for 110 ℃, and mix with other compositions of the bacterium of going out the cooling back, uses as seed culture medium.
The shake flask fermentation substratum
Another substratum that is suitable for cultivating bacterial strain of the present invention is " fermention medium ", and it is a kind of aqueous solution substratum, comprises following composition: 1-10g/L corn slurries, 1-10g/L yeast extract, 5-12g/LKH
2PO
4, 0-3g/L sodium-chlor, 4-12g/L saltpetre, 10-40g/L sucrose, the single alkane of 0.5-3g/L urea and 200-300mL/L or mixed alkanes, 0-1g/L vinylformic acid, 0-0.5g/L tween 80.Described by water and 1N sodium hydroxide solution with pH regulator to 7.5-7.8.With this substratum sterilization 20min under 121 ℃.Urea is sterilized separately, sterilizes 15 minutes down for 110 ℃, and mix with other compositions of the bacterium of going out the cooling back, uses as fermention medium.
According to the present invention, bacterial strain can be used in the various zymotechniques, comprises 500ml fermentation shake flask technology, and it comprises as follows:
With transfering loop uniform spreading on the YPD substratum of the sterilization of cell in the test tube slant is opened.This inclined-plane was cultivated 2 days under stabilizing to 29-30 ℃.Then 1/3rd inoculations of this slant culture are contained in 500ml and shake in the sterilization seed culture medium of the 30ml in the bottle and be 29-30 ℃ in temperature and cultivated 36-48 hour down, bottle speed of shaking is 200-250rpm, and the amplitude of shaking is 2.5-3.5cm.Meat soup with seed fermentation is seeded to the fermention medium that is contained in the sterilization in the 500ml fermentation flask then.This culture is 29-30 ℃ in temperature to be cultivated 90-120 hour down, and bottle speed of shaking is 200-240rpm, and the amplitude of shaking is 2.5-3.5cm, and by monitoring with the adjusting of 1N sodium hydroxide solution pH is maintained between the 7.0-8.0.Dicarboxylic acid concentration in this fermenting broth can be measured after this step.
The evaluation of embodiment 1 candida sake
Screen candida sake (Candida sake) from the oil field, handled the bacterial strain that obtains the positive SL-AH of high yield through chemomorphosis.
The evaluation of candida sake (Candida sake):
1. sample collecting: Accessories during Binzhou suburb, Shandong Province Shengli Oil Field is gathered, and is chocolate crude oil pollution soil sample.
2. the strain separating of collect specimen: it is 10 that all soil samples that will collect are made extent of dilution respectively
-3, 10
-4, 10
-5, 10
-6Diluent be coated with at the YPD flat board respectively.Culture condition: 30 ℃, 48 hours.
Cultivate through separating for several times, obtain oil field saccharomycetic purifying list bacterium colony.
3. identify
With " saccharomycetic means of taxonomic research " (The Yeasts:A Taxonomic Study) the 90th chapter mycocandida (Candida Berkhout) 90.241 candida sakes (Candida sake) comparisons (P1209), determine it is candida sake.
3.1 the utilization of carbon source and nitrogenous source
Under known microorganism identification, carry out nutrition utilization experiment.Add following nitrogenous source or carbon source in defined medium, whether yeast culture is grown to Rule of judgment after 2 days or 3 days, judges negative and the positive, and the result is as follows:
Other test: be that carbon source, nitrogenous source are (+) with the N-acetyl-glucosamine
Be (-) down at 37 ℃
Urase is (-)
Identifying this bacterial strain with this is candida sake (Candida sake).
Embodiment 2 candida sake CAT H4012 mutagenesis screening steps
1. a strain is separated the candida sake obtain, it is stand-by to make the preservation of glycerine pipe.
2. get stand-by one of the glycerine pipe bacterial strain that is preserved in cryogenic refrigerator and put room temperature and thaw, the back that thaws is inserted and is equipped with in the 500ml triangular flask of 50mlYPD substratum, cultivates 20-24h in the rotary shaker that 29 ℃ of rotating speeds are 210rpm.
3. add the YPD nutrient solution of above-mentioned 10ml in the aseptic centrifuge tube of 15ml, centrifugal 2 minutes of 2000rpm removes supernatant liquor, the physiological saline centrifuge washing with 0.85% 3 times.
4. remove supernatant liquor, add 2.5% aseptic lithium chloride 10ml, the preparation bacteria suspension.
5. the NTG mutagenic compound that take by weighing 0.08mg add in the aseptic centrifuge tube of 15ml, and add the physiological saline of 10ml 0.85%, mixing.
6. each the 10ml liquid in 3 and 4 is added in the aseptic plate that is placed with the 15mm*3mm stirrer of 90mm, build loam cake.
7. above-mentioned flat board is by handling 2 of refabrication with quadrat method
8. this plate is put on the magnetic stirring apparatus respectively stir process 10,15,20 minutes.
9. each handles dull and stereotyped processing by the following method respectively.
10. above-mentioned treatment solution is got in the aseptic centrifuge tube of 10ml adding 15ml, and centrifugal 2 minutes of 2000rpm removes supernatant liquor, the physiological saline centrifuge washing with 0.85% 3 times.
11. remove supernatant liquor, the physiological saline with 0.85% carries out gradient dilution step by step.
12. drawing 0.15ml bacterium liquid, every extent of dilution carries out the flat board coating.
13. putting 29 ℃ of incubators, the flat board after the coating is inverted cultivation 3-4 days.
14. picking is cultivated ripe single bacterium and is transferred into the 15*150mm test tube slant, puts 29 ℃ of incubators and cultivates 2 days.
Be equipped with in the 250ml triangular flask of 10ml fermention medium 15. get 1/3rd inclined-plane thalline accesses after the slant culture maturation, in the rotary shaker that 29 ℃ of rotating speeds are 210rpm, cultivated 4 days.4 ℃ of refrigerator cold-storages are put on the residue inclined-plane.
16. calculate the alkali consumption that shakes bottle average alkali consumption and screening bacterial strain of control strain, definite consumption buck that will screen bacterial strain is put down, and generally improves more than 10% than primary dcreening operation control strain alkali consumption.
17. the bacterial strain of how much determining to carry out answering again sieve according to the fermentation shake flask alkali consumption.
18. general 100 strains are shaken and are chosen the high bacterial strain of 5 strain alkali consumptions in bottle primary dcreening operation bacterial strain and carry out multiple sieve.
19. choose the primary dcreening operation inclined-plane of multiple sieve bacterial strain correspondence, above-mentionedly connect remaining 2/3rds inclined-planes behind the primary dcreening operation, shovel is got 1/5th inclined-plane thalline switching inclined-plane F2 again, puts 29 ℃ of incubators and cultivates 2 days.The residue inclined-plane continues to put refrigerator cold-storage.
Be equipped with in the 500ml triangular flask of 30ml seed culture medium 20. cultivate whole whole access of ripe inclined-plane thalline, in the rotary shaker that 29 ℃ of rotating speeds are 210rpm, cultivate 44-48h.Simultaneously 1 glycerine pipe bacterium liquid of control strain also all being inserted 1 seed shakes in the bottle.
Be equipped with in the 500ml triangular flask of 15ml fermention medium 21. the seed liquor after the cultivation maturation is drawn the 3ml adding, in the rotary shaker that 29 ℃ of rotating speeds are 210rpm, cultivate 110h and put bottle mensuration.General each seed bottle correspondence connects 2 bottles of fermented liquids.
22. multiple sieve shakes a bottle measuring method: acid base titration
23. through repeatedly screening the bacterial strain that obtains the high yield SL-AH, called after CAT H4012.
According to candida tropicalis diprotic acid pathways metabolism, selected 15 genes relevant with producing diprotic acid, specific as follows: CYP52A12, CYP52A13, CYP52A14, CYP52A15, CYP52A16, CYP52A17, CYP52A18, CYP52A19, CYP52A20, CYP52D2, POX4, POX5, CPR A, CPR B, and CYTb5.Candida sake CAT H4012 is delivered to specialty order-checking mechanism, adopt Illumina platform end pairing sequencing to carry out genome sequencing, and from genome sequence, extract the gene order with these 15 candida tropicalis dna homologs.Find that candida sake CAT H4012CCTCC M of the present invention 2011485 has any different at the homologous gene of following gene and candida tropicalis: CYP52A12, CYP52D2, POX4, POX5 and CYTb5.Fig. 1-Figure 10 is seen in the gene order contrast.
Embodiment 3 n-dodecane hydrocarbon fermentations
Diprotic acid testing method in the fermented product:
The preparation of diprotic acid sample: fermentation ends, in the 500mL triangular flask, with 6mol/L hydrochloric acid soln adjust pH to 3.0, every bottle adds the 120mL ether, shakes 100 times, place more than the 30min, standing demix takes out the 40mL ether extracted liquid, is added in the 100mL beaker, remove ether, obtain white solid.Carry out the mensuration of diprotic acid then.
Determine the length of diprotic acid, or monoprotic acid do not arranged, then need by vapor-phase chromatography GC that after soon product is handled, separate through capillary column, the diprotic acid of different carbon chain lengths and monoprotic acid go out the peak at different time, therefore just can determine final product.For example: chromatographic apparatus model (GC9800): capillary column FFAP 30*0.53mm*0.5um, detector FID determines that final product compares with retention time Rf and standard substance to draw, standard substance are the AR reagent that Sigma or TGI company produce.
The mensuration of diprotic acid output: the white solid that extraction is obtained, add 100mL inner mark solution (tetramethyl ammonium hydroxide solution that contains the 4mg/mL hexanodioic acid), after the whole dissolvings of the white solid that extraction obtains, air inlet phase chromatogram, the area ratio of mark and sample in the record; Take by weighing standard substance (the diprotic acid standard substance of different carbon chain lengths simultaneously, available from Sigma or TGI reagent company) about 0.4000g, add 100mL inner mark solution (tetramethyl ammonium hydroxide solution that contains the 4mg/mL hexanodioic acid), after standard substance all dissolve, air inlet phase chromatogram, the area ratio of record standard product and sample; Calculate and the calculating of area ratio according to the area normalization method in the vapor-phase chromatography, can obtain the diprotic acid content of different carbon chain lengths.Obtain the standard substance of known quality and the area ratio of internal standard substance by vapor-phase chromatography, so obtain area than with the relation of mass ratio.Obtain the area ratio of sample and internal standard substance again by vapor-phase chromatography, according to aforementioned area than and the relation of mass ratio and the quality of internal standard substance, obtain the quality of sample.
The technical process of 500ml shake flask fermentation and description:
Inclined-plane seed culture-shake-flask seed cultivation-shake flask fermentation
The inclined-plane seed culture: the switching inclined-plane, freeze-drying pipe inclined-plane of 4 ℃ of refrigerator preservations, the seed of transferring behind the cultivation 48h in 29 ℃ of incubators shakes bottle, and seed culture is between 29 ℃ of shaking tables, and 230rpm cultivates secondary fermentation in 48 hours.Fermentation shake flask inserts the seed of 3.0mL, after the inoculation, shakes bottle and puts between 29 ℃ of shaking tables, puts bottle behind the 230rpm shaking culture 90-110h and measures the acid amount of producing.
The substratum that uses in the present embodiment is specific as follows:
The inclined-plane seed culture medium:
20g/L glucose, 10g/L yeast extract and 20g/L peptone, and be 2% agar of YPD substratum weight.Add the agar purpose and be to make substratum gel at room temperature.
The shake-flask seed substratum is as follows:
| Title | Proportioning |
| Sucrose | 20g/L |
| Corn steep liquor | 3g/L |
| Yeast extract paste | 5g/L |
| KH 2PO 4 | 8g/L |
| Heavy wax | 50ml/L |
| Urea (sterilization separately) | 3g/L |
Take by weighing various starting material and record the amount that takes by weighing according to seed culture medium proportioning table, taking by weighing the back that finishes dissolves fully with the tap water starting material, the quantitative charger packing, 30mL/ 500mL shakes bottle, packing, after the packing with the 1.0mL liquid-transfering gun toward shaking the heavy wax that bottle adds 1.5ml, 6 layers of gauze and kraft paper are wrapped up back 121 ℃ of 20min that sterilize down, and is standby.
Fermention medium:
Take by weighing various starting material and record the amount that takes by weighing according to the fermention medium proportioning, taking by weighing the back that finishes dissolves starting material with tap water fully, regulate pH to 7.5 with alkali lye, 15mL/ 500mL shakes bottle, add the n-dodecane hydrocarbon in the fermention medium, wrap up back 121 ℃ of 20min that sterilize down with 6 layers of gauze and kraft paper, standby.
500ml shake flask fermentation result:
Single alkane 500ml shake flask fermentation result:
| Bacterial strain | The diprotic acid kind | Produce acid amount g/L |
| CAT H4012 | Positive 12 carbon | 93.75 |
The fermentation of embodiment 4 mixed alkanes
Mixed alkanes is that n-dodecane hydrocarbon, n-tridecane hydrocarbon, n-tetradecane hydrocarbon, Pentadecane hydrocarbon and n-hexadecane hydrocarbon (nC12+nC13+nC14+nC15+nC16) equal-volume mix.Fermention medium is as follows:
Fermenting process, the diprotic acid kind is all identical with embodiment 3 with the output test in the fermented product.
Mixed alkanes 500ml shake flask fermentation result:
Claims (10)
1. candida sake (Candida sake) CAT H4012 preserving number is CCTCC M2011485.
2. prepare the method for diprotic acid, said method comprising the steps of: a. cultivates candida sake CAT H4012 in the substratum that contains at least a nitrogenous source and at least a organic substrates; Reclaim the diprotic acid of cultivating the step from a with b..
3. method as claimed in claim 2, wherein said at least a organic substrates is selected from the alkane with 12 to 16 carbon atoms.
4. method as claimed in claim 2, wherein said at least a organic substrates is selected from the normal alkane with 12 to 16 carbon atoms.
5. method as claimed in claim 2, wherein said at least a organic substrates is the n-dodecane hydrocarbon.
6. method as claimed in claim 2, wherein said diprotic acid at least a or positive 12 carbon that are 12 carbon to the 16-dicarboxylic acid are at least a to the 16-dicarboxylic acid.
7. as each described method among the claim 2-6, wherein said nitrogenous source is organic nitrogen source or inorganic nitrogen-sourced.
8. method as claimed in claim 7, wherein said organic nitrogen source is one or more in yeast extract paste, corn steep liquor, extractum carnis, soybean meal hydrolysate, peptone, urea, each seed amino acid and the peptide.
9. method as claimed in claim 7, wherein said inorganic nitrogen-sourced be in nitrate, ammoniacal liquor, liquefied ammonia, ammonium salt, the nitrite one or more.
10. candida sake is being produced the application of long-chain biatomic acid according to claim 1.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201210030278.4A CN103243032B (en) | 2012-02-10 | 2012-02-10 | Candida sake and fermentation process thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201210030278.4A CN103243032B (en) | 2012-02-10 | 2012-02-10 | Candida sake and fermentation process thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN103243032A true CN103243032A (en) | 2013-08-14 |
| CN103243032B CN103243032B (en) | 2016-01-27 |
Family
ID=48922912
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201210030278.4A Active CN103243032B (en) | 2012-02-10 | 2012-02-10 | Candida sake and fermentation process thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN103243032B (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP3550014A1 (en) * | 2018-04-04 | 2019-10-09 | Shanghai Cathay Biotech R&D Center Ltd. | Directed evolution of cyp52a12 gene and its use in dicarboxylic acid production |
| CN110343675A (en) * | 2018-04-04 | 2019-10-18 | 上海凯赛生物技术研发中心有限公司 | The directed evolution of CYP52A12 gene and its application in binary acid production |
| CN110684784A (en) * | 2018-07-06 | 2020-01-14 | 上海凯赛生物技术股份有限公司 | Long-chain dibasic acid with low content of monobasic acid impurities and production method thereof |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1233658A (en) * | 1999-05-28 | 1999-11-03 | 清华大学 | Method for screening candida for high prodn. of long chain diacid |
| 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 |
| CN101225411A (en) * | 2007-11-30 | 2008-07-23 | 中国科学院微生物研究所 | New method for biosynthetic production of mixed long-chain dibasic acid |
| CN102191187A (en) * | 2011-04-01 | 2011-09-21 | 山东华星环保集团有限公司 | Candida lipolytica strain and method for preparing long chain dicarboxylic acid by using candida lipolytica strain |
-
2012
- 2012-02-10 CN CN201210030278.4A patent/CN103243032B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1233658A (en) * | 1999-05-28 | 1999-11-03 | 清华大学 | Method for screening candida for high prodn. of long chain diacid |
| 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 |
| CN101225411A (en) * | 2007-11-30 | 2008-07-23 | 中国科学院微生物研究所 | New method for biosynthetic production of mixed long-chain dibasic acid |
| CN102191187A (en) * | 2011-04-01 | 2011-09-21 | 山东华星环保集团有限公司 | Candida lipolytica strain and method for preparing long chain dicarboxylic acid by using candida lipolytica strain |
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP3550014A1 (en) * | 2018-04-04 | 2019-10-09 | Shanghai Cathay Biotech R&D Center Ltd. | Directed evolution of cyp52a12 gene and its use in dicarboxylic acid production |
| CN110343675A (en) * | 2018-04-04 | 2019-10-18 | 上海凯赛生物技术研发中心有限公司 | The directed evolution of CYP52A12 gene and its application in binary acid production |
| US11091742B2 (en) | 2018-04-04 | 2021-08-17 | Cathay Biotech Inc. | Directed evolution of CYP52A12 gene and its use in dicarboxylic acid production |
| CN110343675B (en) * | 2018-04-04 | 2023-05-12 | 上海凯赛生物技术股份有限公司 | Directed evolution of CYP52A12 gene and application thereof in dibasic acid production |
| US11753629B2 (en) | 2018-04-04 | 2023-09-12 | Cathay Biotech Inc. | Directed evolution of CYP52A12 gene and its use in dicarboxylic acid production |
| CN110684784A (en) * | 2018-07-06 | 2020-01-14 | 上海凯赛生物技术股份有限公司 | Long-chain dibasic acid with low content of monobasic acid impurities and production method thereof |
| CN110684784B (en) * | 2018-07-06 | 2023-08-08 | 上海凯赛生物技术股份有限公司 | Long-chain dibasic acid with low content of monobasic acid impurity and production method thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN103243032B (en) | 2016-01-27 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN105112476A (en) | Method for producing lipopeptide biosurfactant through fermentation | |
| CN110438028A (en) | A kind of people pig source Bei Laisi bacillus GX-1 of degraded cellulose | |
| CN102994402B (en) | Strain for producing binary acids and fermentation method of strain | |
| CN104762238A (en) | Lactic acid bacteria not generating amino acid decarboxylase high-yield urease and application of lactic acid bacteria | |
| CN103045494A (en) | Pichia pastoris for efficiently converting methanol to produce single cell protein and application of pichia pastoris | |
| CN105614753A (en) | Micromolecule fish protein nutrition powder and manufacturing method thereof | |
| CN110272835A (en) | One Accharomyces cerevisiae Saccharomyces cerevisiae ScEy01 and application | |
| CN103243032B (en) | Candida sake and fermentation process thereof | |
| RU2706074C1 (en) | Methylococcus capsulatus concept-8 bacteria strain - producer of protein biomass | |
| CN102433288B (en) | Strain for producing ornithine and method for biologically synthesizing ornithine with same | |
| CN113046253B (en) | Culture method for improving heat resistance of kluyveromyces marxianus | |
| CN100558884C (en) | A kind of acid-producing Klebsiella bacterium and application thereof | |
| CN101298623B (en) | Fermentation production method of validacin | |
| CN106544301B (en) | The bacillus licheniformis of one plant of high temperature resistant degrading crude oil galactopoiesis agent and its application | |
| CN105838652B (en) | One plant of bacterial strain for strengthening glycerol metabolism and its application | |
| CN103243034B (en) | Candida sake and fermentation process thereof | |
| CN103243033B (en) | Candida sake and fermentation process thereof | |
| CN110241059A (en) | One plant of low temperature fiber element bacterium for degrading | |
| CN102115765A (en) | Method for producing heptadecanedioic acid by fermenting and converting n-heptadecane | |
| CN103243035B (en) | Candida sake and fermentation process thereof | |
| CN109055284A (en) | A kind of the wine brewing strain of ocean acid-producing bacteria and its application | |
| CN112646740A (en) | Formate single-cell protein strain MA5 and application thereof | |
| CN101407773B (en) | Pseudomonas putida GNA5 bacterial strain and method for preparing D-glucosaminicacid using the same | |
| CN110305802A (en) | The culture medium and cultural method of high spore output monascus purpureus are bred in a kind of liquid state fermentation | |
| CN1854306B (en) | Production of recombinant human serum albumin HSA by fermentation |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C41 | Transfer of patent application or patent right or utility model | ||
| TA01 | Transfer of patent application right |
Effective date of registration: 20151106 Address after: 201203 Shanghai Zhangjiang hi tech park, 1690 No. 5 Cailun Road, building 4 floor Applicant after: CATHAY R&D CENTER Co.,Ltd. Applicant after: CATHAY INDUSTRIAL BIOTECH Ltd. Address before: 201203 Shanghai Zhangjiang hi tech park, 1690 No. 5 Cailun Road, building 4 floor Applicant before: CATHAY R&D CENTER Co.,Ltd. Applicant before: SHANDONG CATHAY BIOTECHNOLOGICAL MATERIAL Co.,Ltd. Applicant before: SHANGHAI CATHAY BIOTECHNOLOGY Co.,Ltd. Applicant before: CATHAY INDUSTRIAL BIOTECH Ltd. |
|
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| TR01 | Transfer of patent right | ||
| TR01 | Transfer of patent right |
Effective date of registration: 20190417 Address after: Fourth Floor, Building 5, No. 1690 Cailun Road, Zhangjiang High-tech Park, Pudong New Area, Shanghai Co-patentee after: CIBT USA Patentee after: CATHAY R&D CENTER Co.,Ltd. Address before: 201203 4th Floor, 5th Building, 1690 Cailun Road, Zhangjiang High-tech Park, Shanghai Co-patentee before: CATHAY INDUSTRIAL BIOTECH Ltd. Patentee before: CATHAY R&D CENTER Co.,Ltd. |
|
| TR01 | Transfer of patent right |
Effective date of registration: 20190531 Address after: Fourth Floor, Building 5, No. 1690 Cailun Road, Zhangjiang High-tech Park, Pudong New Area, Shanghai Co-patentee after: CIBT USA Patentee after: CATHAY R&D CENTER Co.,Ltd. Co-patentee after: Kaisai (Wusu) Biotechnology Co.,Ltd. Address before: 201203 Shanghai Pudong New Area Zhangjiang hi tech park, Cai Lun Road, No. 5, 5 building, 4 floor. Co-patentee before: CIBT USA Patentee before: CATHAY R&D CENTER Co.,Ltd. |
|
| TR01 | Transfer of patent right | ||
| PE01 | Entry into force of the registration of the contract for pledge of patent right |
Denomination of invention: Sake candida and fermentation method thereof Effective date of registration: 20190814 Granted publication date: 20160127 Pledgee: China Import and Export Bank Xinjiang Uygur Autonomous Region Branch Pledgor: Kaisai (Wusu) Biotechnology Co.,Ltd. Registration number: Y2019310000004 |
|
| PE01 | Entry into force of the registration of the contract for pledge of patent right | ||
| CP03 | Change of name, title or address |
Address after: 201203 floor 4, building 5, No. 1690, Cailun Road, Shanghai pilot Free Trade Zone Co-patentee after: CIBT USA Patentee after: CATHAY R&D CENTER Co.,Ltd. Co-patentee after: Kaisai (Wusu) Biotechnology Co.,Ltd. Address before: 201203 Shanghai Zhangjiang High Tech Park of Pudong New Area Cailun Road No. 1690 Building 5 Floor 4 Co-patentee before: CIBT USA Patentee before: CATHAY R&D CENTER Co.,Ltd. Co-patentee before: Kaisai (Wusu) Biotechnology Co.,Ltd. |
|
| CP03 | Change of name, title or address | ||
| PC01 | Cancellation of the registration of the contract for pledge of patent right |
Date of cancellation: 20230717 Granted publication date: 20160127 Pledgee: China Import and Export Bank Xinjiang Uygur Autonomous Region Branch Pledgor: Kaisai (Wusu) Biotechnology Co.,Ltd. Registration number: Y2019310000004 |
|
| PC01 | Cancellation of the registration of the contract for pledge of patent right |