JP3354608B2 - Method for producing polyunsaturated fatty acid and lipid containing the same - Google Patents
Method for producing polyunsaturated fatty acid and lipid containing the sameInfo
- Publication number
- JP3354608B2 JP3354608B2 JP30552392A JP30552392A JP3354608B2 JP 3354608 B2 JP3354608 B2 JP 3354608B2 JP 30552392 A JP30552392 A JP 30552392A JP 30552392 A JP30552392 A JP 30552392A JP 3354608 B2 JP3354608 B2 JP 3354608B2
- Authority
- JP
- Japan
- Prior art keywords
- culture
- ppm
- fatty acid
- mortierella
- producing
- 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.)
- Expired - Lifetime
Links
- 235000020777 polyunsaturated fatty acids Nutrition 0.000 title claims description 23
- 238000004519 manufacturing process Methods 0.000 title claims description 20
- 150000002632 lipids Chemical class 0.000 title claims description 11
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 claims description 33
- 239000001301 oxygen Substances 0.000 claims description 33
- 229910052760 oxygen Inorganic materials 0.000 claims description 33
- 238000000034 method Methods 0.000 claims description 25
- 241000235575 Mortierella Species 0.000 claims description 17
- 244000005700 microbiome Species 0.000 claims description 16
- 238000005273 aeration Methods 0.000 claims description 11
- 238000012258 culturing Methods 0.000 claims description 10
- 239000007788 liquid Substances 0.000 claims description 3
- 235000021122 unsaturated fatty acids Nutrition 0.000 claims description 2
- 150000004670 unsaturated fatty acids Chemical class 0.000 claims description 2
- HOBAELRKJCKHQD-UHFFFAOYSA-N (8Z,11Z,14Z)-8,11,14-eicosatrienoic acid Natural products CCCCCC=CCC=CCC=CCCCCCCC(O)=O HOBAELRKJCKHQD-UHFFFAOYSA-N 0.000 description 23
- HOBAELRKJCKHQD-QNEBEIHSSA-N dihomo-γ-linolenic acid Chemical compound CCCCC\C=C/C\C=C/C\C=C/CCCCCCC(O)=O HOBAELRKJCKHQD-QNEBEIHSSA-N 0.000 description 23
- PYMYPHUHKUWMLA-UHFFFAOYSA-N arabinose Natural products OCC(O)C(O)C(O)C=O PYMYPHUHKUWMLA-UHFFFAOYSA-N 0.000 description 22
- SRBFZHDQGSBBOR-UHFFFAOYSA-N beta-D-Pyranose-Lyxose Natural products OC1COC(O)C(O)C1O SRBFZHDQGSBBOR-UHFFFAOYSA-N 0.000 description 22
- 210000004027 cell Anatomy 0.000 description 18
- 235000014113 dietary fatty acids Nutrition 0.000 description 18
- 229930195729 fatty acid Natural products 0.000 description 18
- 239000000194 fatty acid Substances 0.000 description 18
- 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 13
- 239000008103 glucose Substances 0.000 description 13
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 12
- 150000004665 fatty acids Chemical class 0.000 description 10
- 241000233866 Fungi Species 0.000 description 7
- 229940041514 candida albicans extract Drugs 0.000 description 7
- 239000012138 yeast extract Substances 0.000 description 7
- 239000002518 antifoaming agent Substances 0.000 description 6
- 238000001816 cooling Methods 0.000 description 6
- 230000000694 effects Effects 0.000 description 6
- 238000004817 gas chromatography Methods 0.000 description 6
- 229910052757 nitrogen Inorganic materials 0.000 description 6
- 238000009423 ventilation Methods 0.000 description 6
- 241001219224 Mortierella elongata Species 0.000 description 5
- 241000048020 Mortierella exigua Species 0.000 description 5
- 239000004006 olive oil Substances 0.000 description 5
- 235000008390 olive oil Nutrition 0.000 description 5
- 238000003756 stirring Methods 0.000 description 5
- 241000907999 Mortierella alpina Species 0.000 description 4
- 239000007789 gas Substances 0.000 description 4
- 230000001954 sterilising effect Effects 0.000 description 4
- 241000894006 Bacteria Species 0.000 description 3
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical group [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 3
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 3
- 235000020673 eicosapentaenoic acid Nutrition 0.000 description 3
- SRBFZHDQGSBBOR-IOVATXLUSA-N D-xylopyranose Chemical compound O[C@@H]1COC(O)[C@H](O)[C@H]1O SRBFZHDQGSBBOR-IOVATXLUSA-N 0.000 description 2
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 2
- 108090000790 Enzymes Proteins 0.000 description 2
- 102000004190 Enzymes Human genes 0.000 description 2
- OAKJQQAXSVQMHS-UHFFFAOYSA-N Hydrazine Chemical compound NN OAKJQQAXSVQMHS-UHFFFAOYSA-N 0.000 description 2
- 241000108463 Hygrophila <snail> Species 0.000 description 2
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 description 2
- 241000133355 Mortierella hygrophila Species 0.000 description 2
- NWIBSHFKIJFRCO-WUDYKRTCSA-N Mytomycin Chemical compound C1N2C(C(C(C)=C(N)C3=O)=O)=C3[C@@H](COC(N)=O)[C@@]2(OC)[C@@H]2[C@H]1N2 NWIBSHFKIJFRCO-WUDYKRTCSA-N 0.000 description 2
- ISWSIDIOOBJBQZ-UHFFFAOYSA-N Phenol Chemical compound OC1=CC=CC=C1 ISWSIDIOOBJBQZ-UHFFFAOYSA-N 0.000 description 2
- XSQUKJJJFZCRTK-UHFFFAOYSA-N Urea Chemical compound NC(N)=O XSQUKJJJFZCRTK-UHFFFAOYSA-N 0.000 description 2
- VEXNFKCQMGMBBJ-UHFFFAOYSA-N [1-(dimethylamino)-2-[(dimethylamino)methyl]butan-2-yl] benzoate Chemical compound CN(C)CC(CC)(CN(C)C)OC(=O)C1=CC=CC=C1 VEXNFKCQMGMBBJ-UHFFFAOYSA-N 0.000 description 2
- YZXBAPSDXZZRGB-DOFZRALJSA-N arachidonic acid Chemical compound CCCCC\C=C/C\C=C/C\C=C/C\C=C/CCCC(O)=O YZXBAPSDXZZRGB-DOFZRALJSA-N 0.000 description 2
- 229910052799 carbon Inorganic materials 0.000 description 2
- 125000004432 carbon atom Chemical group C* 0.000 description 2
- 238000006243 chemical reaction Methods 0.000 description 2
- 239000000306 component Substances 0.000 description 2
- 230000003247 decreasing effect Effects 0.000 description 2
- 238000000855 fermentation Methods 0.000 description 2
- 230000004151 fermentation Effects 0.000 description 2
- DCAYPVUWAIABOU-UHFFFAOYSA-N hexadecane Chemical compound CCCCCCCCCCCCCCCC DCAYPVUWAIABOU-UHFFFAOYSA-N 0.000 description 2
- IPCSVZSSVZVIGE-UHFFFAOYSA-N hexadecanoic acid Chemical compound CCCCCCCCCCCCCCCC(O)=O IPCSVZSSVZVIGE-UHFFFAOYSA-N 0.000 description 2
- 239000003112 inhibitor Substances 0.000 description 2
- GLVAUDGFNGKCSF-UHFFFAOYSA-N mercaptopurine Chemical compound S=C1NC=NC2=C1NC=N2 GLVAUDGFNGKCSF-UHFFFAOYSA-N 0.000 description 2
- MBABOKRGFJTBAE-UHFFFAOYSA-N methyl methanesulfonate Chemical compound COS(C)(=O)=O MBABOKRGFJTBAE-UHFFFAOYSA-N 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 235000015097 nutrients Nutrition 0.000 description 2
- RZJRJXONCZWCBN-UHFFFAOYSA-N octadecane Chemical compound CCCCCCCCCCCCCCCCCC RZJRJXONCZWCBN-UHFFFAOYSA-N 0.000 description 2
- 235000020660 omega-3 fatty acid Nutrition 0.000 description 2
- BASFCYQUMIYNBI-UHFFFAOYSA-N platinum Chemical compound [Pt] BASFCYQUMIYNBI-UHFFFAOYSA-N 0.000 description 2
- KMUONIBRACKNSN-UHFFFAOYSA-N potassium dichromate Chemical compound [K+].[K+].[O-][Cr](=O)(=O)O[Cr]([O-])(=O)=O KMUONIBRACKNSN-UHFFFAOYSA-N 0.000 description 2
- 230000005855 radiation Effects 0.000 description 2
- 150000003839 salts Chemical class 0.000 description 2
- VWDWKYIASSYTQR-UHFFFAOYSA-N sodium nitrate Chemical compound [Na+].[O-][N+]([O-])=O VWDWKYIASSYTQR-UHFFFAOYSA-N 0.000 description 2
- 239000000758 substrate Substances 0.000 description 2
- BGHCVCJVXZWKCC-UHFFFAOYSA-N tetradecane Chemical compound CCCCCCCCCCCCCC BGHCVCJVXZWKCC-UHFFFAOYSA-N 0.000 description 2
- UNSRRHDPHVZAHH-YOILPLPUSA-N (5Z,8Z,11Z)-icosatrienoic acid Chemical compound CCCCCCCC\C=C/C\C=C/C\C=C/CCCC(O)=O UNSRRHDPHVZAHH-YOILPLPUSA-N 0.000 description 1
- UYEILDGRSAARCQ-UHFFFAOYSA-N 1-methyl-1,2-dinitrosoguanidine Chemical compound O=NN(C)C(=N)NN=O UYEILDGRSAARCQ-UHFFFAOYSA-N 0.000 description 1
- OWEGMIWEEQEYGQ-UHFFFAOYSA-N 100676-05-9 Natural products OC1C(O)C(O)C(CO)OC1OCC1C(O)C(O)C(O)C(OC2C(OC(O)C(O)C2O)CO)O1 OWEGMIWEEQEYGQ-UHFFFAOYSA-N 0.000 description 1
- PAWQVTBBRAZDMG-UHFFFAOYSA-N 2-(3-bromo-2-fluorophenyl)acetic acid Chemical compound OC(=O)CC1=CC=CC(Br)=C1F PAWQVTBBRAZDMG-UHFFFAOYSA-N 0.000 description 1
- ZPVSFHWMXABGPU-UHFFFAOYSA-N 4-nitroquinoline Chemical compound C1=CC=C2C([N+](=O)[O-])=CC=NC2=C1 ZPVSFHWMXABGPU-UHFFFAOYSA-N 0.000 description 1
- LQLQRFGHAALLLE-UHFFFAOYSA-N 5-bromouracil Chemical compound BrC1=CNC(=O)NC1=O LQLQRFGHAALLLE-UHFFFAOYSA-N 0.000 description 1
- UNSRRHDPHVZAHH-UHFFFAOYSA-N 6beta,11alpha-Dihydroxy-3alpha,5alpha-cyclopregnan-20-on Natural products CCCCCCCCC=CCC=CCC=CCCCC(O)=O UNSRRHDPHVZAHH-UHFFFAOYSA-N 0.000 description 1
- 102100034542 Acyl-CoA (8-3)-desaturase Human genes 0.000 description 1
- 229920001817 Agar Polymers 0.000 description 1
- 239000004215 Carbon black (E152) Substances 0.000 description 1
- FBPFZTCFMRRESA-KVTDHHQDSA-N D-Mannitol Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-KVTDHHQDSA-N 0.000 description 1
- 108010073542 Delta-5 Fatty Acid Desaturase Proteins 0.000 description 1
- MYMOFIZGZYHOMD-UHFFFAOYSA-N Dioxygen Chemical compound O=O MYMOFIZGZYHOMD-UHFFFAOYSA-N 0.000 description 1
- 229930091371 Fructose Natural products 0.000 description 1
- RFSUNEUAIZKAJO-ARQDHWQXSA-N Fructose Chemical compound OC[C@H]1O[C@](O)(CO)[C@@H](O)[C@@H]1O RFSUNEUAIZKAJO-ARQDHWQXSA-N 0.000 description 1
- 239000005715 Fructose Substances 0.000 description 1
- AVXURJPOCDRRFD-UHFFFAOYSA-N Hydroxylamine Chemical compound ON AVXURJPOCDRRFD-UHFFFAOYSA-N 0.000 description 1
- SRBFZHDQGSBBOR-HWQSCIPKSA-N L-arabinopyranose Chemical compound O[C@H]1COC(O)[C@H](O)[C@H]1O SRBFZHDQGSBBOR-HWQSCIPKSA-N 0.000 description 1
- GUBGYTABKSRVRQ-PICCSMPSSA-N Maltose Natural products O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@@H](CO)OC(O)[C@H](O)[C@H]1O GUBGYTABKSRVRQ-PICCSMPSSA-N 0.000 description 1
- 229910021380 Manganese Chloride Inorganic materials 0.000 description 1
- GLFNIEUTAYBVOC-UHFFFAOYSA-L Manganese chloride Chemical compound Cl[Mn]Cl GLFNIEUTAYBVOC-UHFFFAOYSA-L 0.000 description 1
- 229930195725 Mannitol Natural products 0.000 description 1
- 241001558147 Mortierella sp. Species 0.000 description 1
- TUNFSRHWOTWDNC-UHFFFAOYSA-N Myristic acid Natural products CCCCCCCCCCCCCC(O)=O TUNFSRHWOTWDNC-UHFFFAOYSA-N 0.000 description 1
- IOVCWXUNBOPUCH-UHFFFAOYSA-N Nitrous acid Chemical compound ON=O IOVCWXUNBOPUCH-UHFFFAOYSA-N 0.000 description 1
- 229910019142 PO4 Inorganic materials 0.000 description 1
- 235000021314 Palmitic acid Nutrition 0.000 description 1
- 239000001888 Peptone Substances 0.000 description 1
- 108010080698 Peptones Proteins 0.000 description 1
- 101100364827 Prochlorococcus marinus (strain SARG / CCMP1375 / SS120) ahcY gene Proteins 0.000 description 1
- WDVSHHCDHLJJJR-UHFFFAOYSA-N Proflavine Chemical compound C1=CC(N)=CC2=NC3=CC(N)=CC=C3C=C21 WDVSHHCDHLJJJR-UHFFFAOYSA-N 0.000 description 1
- 101150081953 SAM1 gene Proteins 0.000 description 1
- 108010073771 Soybean Proteins Proteins 0.000 description 1
- 229920002472 Starch Polymers 0.000 description 1
- 235000021355 Stearic acid Nutrition 0.000 description 1
- 229930006000 Sucrose Natural products 0.000 description 1
- 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 1
- 240000008042 Zea mays Species 0.000 description 1
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 description 1
- 235000002017 Zea mays subsp mays Nutrition 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 239000012190 activator Substances 0.000 description 1
- 238000007259 addition reaction Methods 0.000 description 1
- 239000008272 agar Substances 0.000 description 1
- JAZBEHYOTPTENJ-JLNKQSITSA-N all-cis-5,8,11,14,17-icosapentaenoic acid Chemical compound CC\C=C/C\C=C/C\C=C/C\C=C/C\C=C/CCCC(O)=O JAZBEHYOTPTENJ-JLNKQSITSA-N 0.000 description 1
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 description 1
- 229910052921 ammonium sulfate Inorganic materials 0.000 description 1
- 235000011130 ammonium sulphate Nutrition 0.000 description 1
- 239000003242 anti bacterial agent Substances 0.000 description 1
- 235000021342 arachidonic acid Nutrition 0.000 description 1
- 229940114079 arachidonic acid Drugs 0.000 description 1
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 description 1
- GUBGYTABKSRVRQ-QUYVBRFLSA-N beta-maltose Chemical compound OC[C@H]1O[C@H](O[C@H]2[C@H](O)[C@@H](O)[C@H](O)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@@H]1O GUBGYTABKSRVRQ-QUYVBRFLSA-N 0.000 description 1
- 230000003115 biocidal effect Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 239000004202 carbamide Substances 0.000 description 1
- 231100000357 carcinogen Toxicity 0.000 description 1
- 239000003183 carcinogenic agent Substances 0.000 description 1
- SIHHLZPXQLFPMC-UHFFFAOYSA-N chloroform;methanol;hydrate Chemical compound O.OC.ClC(Cl)Cl SIHHLZPXQLFPMC-UHFFFAOYSA-N 0.000 description 1
- 239000003240 coconut oil Substances 0.000 description 1
- 235000019864 coconut oil Nutrition 0.000 description 1
- 229910000365 copper sulfate Inorganic materials 0.000 description 1
- ARUVKPQLZAKDPS-UHFFFAOYSA-L copper(II) sulfate Chemical compound [Cu+2].[O-][S+2]([O-])([O-])[O-] ARUVKPQLZAKDPS-UHFFFAOYSA-L 0.000 description 1
- 235000005822 corn Nutrition 0.000 description 1
- 235000012343 cottonseed oil Nutrition 0.000 description 1
- 239000002385 cottonseed oil Substances 0.000 description 1
- 210000004748 cultured cell Anatomy 0.000 description 1
- 235000013681 dietary sucrose Nutrition 0.000 description 1
- 238000001035 drying Methods 0.000 description 1
- 229960005135 eicosapentaenoic acid Drugs 0.000 description 1
- JAZBEHYOTPTENJ-UHFFFAOYSA-N eicosapentaenoic acid Natural products CCC=CCC=CCC=CCC=CCC=CCCCC(O)=O JAZBEHYOTPTENJ-UHFFFAOYSA-N 0.000 description 1
- 230000032050 esterification Effects 0.000 description 1
- 238000005886 esterification reaction Methods 0.000 description 1
- 239000000284 extract Substances 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 239000003925 fat Substances 0.000 description 1
- 235000019197 fats Nutrition 0.000 description 1
- -1 fatty acid ester Chemical class 0.000 description 1
- 238000012262 fermentative production Methods 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- VZCCETWTMQHEPK-UHFFFAOYSA-N gamma-Linolensaeure Natural products CCCCCC=CCC=CCC=CCCCCC(O)=O VZCCETWTMQHEPK-UHFFFAOYSA-N 0.000 description 1
- VZCCETWTMQHEPK-QNEBEIHSSA-N gamma-linolenic acid Chemical compound CCCCC\C=C/C\C=C/C\C=C/CCCCC(O)=O VZCCETWTMQHEPK-QNEBEIHSSA-N 0.000 description 1
- 235000020664 gamma-linolenic acid Nutrition 0.000 description 1
- 229960002733 gamolenic acid Drugs 0.000 description 1
- 235000011187 glycerol Nutrition 0.000 description 1
- 238000010438 heat treatment Methods 0.000 description 1
- 229930195733 hydrocarbon Natural products 0.000 description 1
- 150000002430 hydrocarbons Chemical class 0.000 description 1
- FUKUFMFMCZIRNT-UHFFFAOYSA-N hydron;methanol;chloride Chemical compound Cl.OC FUKUFMFMCZIRNT-UHFFFAOYSA-N 0.000 description 1
- 229910000358 iron sulfate Inorganic materials 0.000 description 1
- BAUYGSIQEAFULO-UHFFFAOYSA-L iron(2+) sulfate (anhydrous) Chemical compound [Fe+2].[O-]S([O-])(=O)=O BAUYGSIQEAFULO-UHFFFAOYSA-L 0.000 description 1
- 238000009630 liquid culture Methods 0.000 description 1
- 229910052943 magnesium sulfate Inorganic materials 0.000 description 1
- 235000019341 magnesium sulphate Nutrition 0.000 description 1
- 235000002867 manganese chloride Nutrition 0.000 description 1
- 239000011565 manganese chloride Substances 0.000 description 1
- 229940099607 manganese chloride Drugs 0.000 description 1
- 239000000594 mannitol Substances 0.000 description 1
- 235000010355 mannitol Nutrition 0.000 description 1
- 235000013372 meat Nutrition 0.000 description 1
- HAWPXGHAZFHHAD-UHFFFAOYSA-N mechlorethamine Chemical class ClCCN(C)CCCl HAWPXGHAZFHHAD-UHFFFAOYSA-N 0.000 description 1
- 229960004961 mechlorethamine Drugs 0.000 description 1
- 239000012533 medium component Substances 0.000 description 1
- 229960001428 mercaptopurine Drugs 0.000 description 1
- WSFSSNUMVMOOMR-NJFSPNSNSA-N methanone Chemical compound O=[14CH2] WSFSSNUMVMOOMR-NJFSPNSNSA-N 0.000 description 1
- 125000002496 methyl group Chemical group [H]C([H])([H])* 0.000 description 1
- 229960004857 mitomycin Drugs 0.000 description 1
- 235000013379 molasses Nutrition 0.000 description 1
- 239000003471 mutagenic agent Substances 0.000 description 1
- 231100000707 mutagenic chemical Toxicity 0.000 description 1
- 230000003505 mutagenic effect Effects 0.000 description 1
- 230000035772 mutation Effects 0.000 description 1
- IAIWVQXQOWNYOU-FPYGCLRLSA-N nitrofural Chemical class NC(=O)N\N=C\C1=CC=C([N+]([O-])=O)O1 IAIWVQXQOWNYOU-FPYGCLRLSA-N 0.000 description 1
- 229940038384 octadecane Drugs 0.000 description 1
- QIQXTHQIDYTFRH-UHFFFAOYSA-N octadecanoic acid Chemical compound CCCCCCCCCCCCCCCCCC(O)=O QIQXTHQIDYTFRH-UHFFFAOYSA-N 0.000 description 1
- OQCDKBAXFALNLD-UHFFFAOYSA-N octadecanoic acid Natural products CCCCCCCC(C)CCCCCCCCC(O)=O OQCDKBAXFALNLD-UHFFFAOYSA-N 0.000 description 1
- 239000003921 oil Substances 0.000 description 1
- 235000019198 oils Nutrition 0.000 description 1
- 235000020665 omega-6 fatty acid Nutrition 0.000 description 1
- 125000001477 organic nitrogen group Chemical group 0.000 description 1
- 235000019319 peptone Nutrition 0.000 description 1
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 description 1
- 239000010452 phosphate Substances 0.000 description 1
- 229910052697 platinum Inorganic materials 0.000 description 1
- 229960000286 proflavine Drugs 0.000 description 1
- 230000035755 proliferation Effects 0.000 description 1
- 229920006395 saturated elastomer Polymers 0.000 description 1
- 238000013341 scale-up Methods 0.000 description 1
- 239000004317 sodium nitrate Substances 0.000 description 1
- 235000010344 sodium nitrate Nutrition 0.000 description 1
- 239000002689 soil Substances 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 239000003549 soybean oil Substances 0.000 description 1
- 235000012424 soybean oil Nutrition 0.000 description 1
- 235000019710 soybean protein Nutrition 0.000 description 1
- 239000008107 starch Substances 0.000 description 1
- 235000019698 starch Nutrition 0.000 description 1
- 238000004659 sterilization and disinfection Methods 0.000 description 1
- 229960004793 sucrose Drugs 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 239000013076 target substance Substances 0.000 description 1
- 235000013343 vitamin Nutrition 0.000 description 1
- 239000011782 vitamin Substances 0.000 description 1
- 229940088594 vitamin Drugs 0.000 description 1
- 229930003231 vitamin Natural products 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
Landscapes
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Description
【０００１】[0001]
【産業上の利用分野】本発明は、モルティエレラ（Ｍｏ
ｒｔｉｅｒｅｌｌａ）属のモルティエレラ（Ｍｏｒｔｉ
ｅｒｅｌｌａ）亜属に属する微生物を利用した発酵法に
よる、ω−３系やω−６系、ω−９系等の高度不飽和脂
肪酸（以下ＰＵＦＡとする）又はこれらを含有する脂質
を製造する方法に関する。BACKGROUND OF THE INVENTION The present invention relates to Mortierella ( Mo).
rtierella) of the genus Mortierella (Morti
A method for producing highly unsaturated fatty acids (hereinafter referred to as PUFA) such as ω-3, ω-6, and ω-9, or lipids containing these, by a fermentation method using a microorganism belonging to the subgenus erella ) About.
【０００２】糸状菌であるモルティエレラ属微生物を用
いてアラキドン酸（以下、ＡＲＡと称する）やジホモ−
γ−リノレン酸（以下、ＤＧＬＡと称する）をはじめと
するω−６系ＰＵＦＡを製造することは既に知られてい
る（特開昭６３−０４４８９１、特開平１−２４３９９
２）。又、モルティエレラ属微生物を用いて、低温培養
することによりω−３系ＰＵＦＡであるエイコサペンタ
エン酸（以下ＥＰＡとする）を製造することも知られて
いる（特開昭６３−１４６９７）。さらにモルティエレ
ラ属微生物の突然変異株を用いて、ミード酸等のω−９
系ＰＵＦＡを製造することやＤＧＬＡを製造することが
見い出されている（特願平３−２５１９６６、特願平３
−２５１９６４）。[0002] Arachidonic acid (hereinafter referred to as ARA) or dihomo-
It is already known to produce ω-6-based PUFAs such as γ-linolenic acid (hereinafter referred to as DGLA) (JP-A-63-044891, JP-A-1-24399).
2). It is also known to produce eicosapentaenoic acid (hereinafter referred to as EPA) which is a ω-3 PUFA by culturing at a low temperature using a microorganism of the genus Mortierella (JP-A-63-14697). Further, using a mutant strain of Mortierella microorganism, ω-9 such as mead acid is used.
It has been found that a system-based PUFA is manufactured or a DGLA is manufactured (Japanese Patent Application Nos. 3-251966, 3).
251964).
【０００３】これらの脂肪酸が該菌体内で生産される際
の菌体内での脂肪酸不飽和化反応は、酸素添加反応によ
る好気的不飽和化反応であり、培養液中の溶存酸素濃度
（以下ＤＯとする）はＰＵＦＡ生産における重要な因子
と考えられる。糸状菌によるＰＵＦＡ生産に及ぼすＤＯ
の影響については、２〜３ppm の比較的低いＤＯの範囲
内で検討された報告はあるが（Appl. Microbiol. Biote
chnol., 37, 18 (1992))、常圧下での通常空気を通気し
た場合の飽和ＤＯに近い値、またはそれを越える高いＤ
Ｏ条件下などでの糸状菌によるＰＵＦＡ生産について詳
細に検討された報告はない。[0003] The fatty acid desaturation reaction in the cells when these fatty acids are produced in the cells is an aerobic desaturation reaction due to an oxygen addition reaction, and the dissolved oxygen concentration in the culture solution (hereinafter referred to as "the concentration of dissolved oxygen"). DO) is considered an important factor in PUFA production. Effect of DO on PUFA production by filamentous fungi
Although the effect of phenol has been studied in the range of a relatively low DO of 2-3 ppm (Appl. Microbiol. Biote.
chnol., 37 , 18 (1992)), a value close to or higher than the saturated DO when normal air is ventilated under normal pressure.
There is no report that examined the production of PUFA by filamentous fungi under O conditions or the like in detail.
【０００４】一方、モルティエレラ属のような糸状菌を
用いて、液体培地で発酵生産を行なう場合、往々にして
菌体増殖による培養液粘度の増加とそれに伴う酸素供給
の不足が起こる（Biotechnol. Bioeng.,37, 960 (199
1)) 。その対策として、一般的には培養槽の攪拌回転数
を上げる等の手段が知られているが、それは一般にせん
断力に弱いと言われている糸状菌の培養には適しておら
ず、このことが糸状菌のスケールアップに伴う生産性低
下の一因となっている（J. Ferment. Technol.,56, 374
(1978), Biotechnol. Bioeng.,35, 1011 (1990))。On the other hand, when fermentative production is performed in a liquid medium using a filamentous fungus such as Mortierella sp., The viscosity of the culture solution often increases due to the proliferation of cells, and the resulting lack of oxygen supply occurs (Biotechnol. Bioeng., 37 , 960 (199
1)). As a countermeasure, generally known means such as increasing the number of rotations of the culture tank, but this is not suitable for culturing filamentous fungi, which is generally said to be weak against shear force. Contribute to a decrease in productivity due to the scale-up of filamentous fungi (J. Ferment. Technol., 56 , 374).
(1978), Biotechnol. Bioeng., 35 , 1011 (1990)).
【０００５】このような糸状菌に対して酸素供給を十分
に行なう方策として、培養槽の形状改良の試み（Biotec
hnol. Lett.,14, 491 (1992), Appl. Microbiol. Biote
chnol., 37, 32 (1992), Appl. Microbiol. Biotechno
l.,37, 37 (1992), Biotechnol. Bioeng., 32, 835 (19
88)) などが成されているが、多目的に使用される工業
規模での培養層においては、その形状変更は実用上不可
能である。As a measure to sufficiently supply oxygen to such filamentous fungi, an attempt to improve the shape of a culture tank (Biotec
hnol. Lett., 14 , 491 (1992), Appl. Microbiol. Biote
chnol., 37 , 32 (1992), Appl. Microbiol. Biotechno
l., 37 , 37 (1992), Biotechnol. Bioeng., 32 , 835 (19
88)), etc., but it is practically impossible to change the shape of the culture layer on an industrial scale used for multiple purposes.
【０００６】[0006]
【発明が解決しようとする課題】本発明はモルティエレ
ラ亜属微生物によるＰＵＦＡ生産効率とＤＯの関係を解
明し、微生物に損傷を与えることなく、工業規模でも容
易に実施できる酸素供給方法を用いて同微生物による効
率的なＡＲＡ，ＤＧＬＡをはじめとするＰＵＦＡまたは
これらを含有する脂質の製造方法を提供しようとするも
のである。DISCLOSURE OF THE INVENTION The present invention clarifies the relationship between PUFA production efficiency and DO by Mortierella subgenus microorganisms, and uses an oxygen supply method which can be easily carried out on an industrial scale without damaging the microorganisms. An object of the present invention is to provide a method for efficiently producing PUFAs such as ARA and DGLA or lipids containing these by the microorganism.
【０００７】[0007]
【課題を解決するための手段】本発明者等は上記の課題
を解決するため、ＤＯと目的とするＰＵＦＡの生産量と
の関係を詳細に検討した結果、モルティエレラ亜属の微
生物を用いてＰＵＦＡを製造する場合、液体培養時に、
培養液中のＤＯを５〜２８ppm に維持すれば、目的物で
あるＰＵＦＡを効率的に生産できること、且つ微生物を
損傷することなく、簡単に酸素供給する方法として培養
槽加圧法、および酸素富化空気通気法が有効であること
を見出し、本発明を完成するに至った。Means for Solving the Problems In order to solve the above-mentioned problems, the present inventors have studied in detail the relationship between DO and the target PUFA production, and as a result, using microorganisms of the subgenus Mortierella. When producing PUFA, at the time of liquid culture,
If DO in the culture solution is maintained at 5 to 28 ppm, PUFA, which is the target substance, can be produced efficiently, and the culture tank pressurization method and the oxygen enrichment method can be easily supplied without damaging microorganisms. The inventor found that the air ventilation method was effective, and completed the present invention.
【０００８】[0008]
【具体的な説明】本発明において、モルティエレラ属の
モルティエレラ亜属に属する微生物であれば、すべて使
用することができ、例えば、モルティエレラ・エロンガ
タ（Ｍｏｒｔｉｅｒｅｌｌａ ｅｌｏｎｇａｔａ）、モ
ルティエレラ・エキシグア（Ｍｏｒｔｉｅｒｅｌｌａ
ｅｘｉｇｕａ）、モルティエレラ・ヒグロフィラ（Ｍｏ
ｒｔｉｅｒｅｌｌａ ｈｙｇｒｏｐｈｉｌａ）、モルテ
ィエレラ・アルピナ（Ｍｏｒｔｉｅｒｅｌｌａ ａｌｐ
ｉｎａ）等の菌を挙げることができる。In DETAILED DESCRIPTION The present invention, as long as a microorganism belonging to Mortierella subgenus Mortierella, all can be used, for example, Mortierella elongata (Mortierella elongata), Mortierella exigua (Mortierella
exigua ), Mortierella hygrophylla ( Mo)
rtierella hygrophila), Mortierella alpina (Mortierella alp
ina ).
【０００９】さらに具体的には、モルティエレラ・エロ
ンガタ（Ｍｏｒｔｉｅｒｅｌｌａｅｌｏｎｇａｔａ）Ｉ
ＦＯ８５７０、モルティエレラ・エキシグア（Ｍｏｒｔ
ｉｅｒｅｌｌａ ｅｘｉｇｕａ）ＩＦＯ８５７１、モル
ティエレラ・ヒグロフィラ（Ｍｏｒｔｉｅｒｅｌｌａ
ｈｙｇｒｏｐｈｉｌａ）ＩＦＯ５９４１、モルティエレ
ラ・アルピナ（Ｍｏｒｔｉｅｒｅｌｌａ ａｌｐｉｎ
ａ）ＩＦＯ８５６８等の菌株を挙げることができ、これ
らはいずれも、財団法人醗酵研究所からなんら制限なく
入手することができる。また土壌分離菌株であるモルテ
ィエレラ・エロンガタ（Ｍｏｒｔｉｅｒｅｌｌａ ｅｌ
ｏｎｇａｔａ）ＳＡＭ０２１９（微工研条寄１２３９
号）を使用することもできる。しかしながらこれらの菌
に限定されるものではない。More specifically, Mortierella Ero
Ngata (Mortierellalongagata) I
FO8570, Mortierella Exigua (Mort
ierella exigua) IFO8571, mol
Thierella hygrophila (Mortierella
hygrophila) IFO5941, Mortiere
La Alpina (Mortierella alpin
a) Strains such as IFO8568;
Have no restrictions from the Fermentation Research Institute
Can be obtained. Morte, a soil isolate,
Jeelela Elongata (Mortierella el
ongata) SAM0219 (1239)
No.) can also be used. However, these bacteria
However, the present invention is not limited to this.
【００１０】また本発明において使用できるモルティエ
レラ亜属に属する微生物の中には、その突然変異株も含
まれる。例えばモルティエレラ亜属に属する微生物に突
然変異操作を行ない、不飽和化酵素や炭素鎖延長化酵素
の活性が低下または欠損あるいは向上した変異株を使用
することができる。さらに具体的には、Δ５不飽和化酵
素活性が低下した変異株としてモルティエレラ・アルピ
ナ（Ｍｏｒｔｉｅｒｅｌｌａ ａｌｐｉｎａ）ＳＡＭ１
８６０（微工研条寄３５８９号）、Δ１２不飽和化酵素
活性が低下した変異株としてモルティエレラ・アルピナ
（Ｍｏｒｔｉｅｒｅｌｌａ ａｌｐｉｎａ）ＳＡＭ１８
６１（微工研条寄３５９０号）等の変異株を挙げること
ができる。しかしながらこれらの菌に限定されるもので
はない。The microorganism belonging to the subgenus Mortierella which can be used in the present invention also includes a mutant thereof. For example, a mutant strain in which the activity of a desaturase or a carbon chain elongation enzyme is reduced, deleted, or improved can be used by mutating a microorganism belonging to the subgenus Mortierella. More specifically, Mortierella alpina SAM1 is a mutant strain having a decreased Δ5 desaturase activity.
860 (Microtechnical Research Laboratories No. 3589), Mortierella alpina SAM18 as a mutant strain having reduced Δ12 desaturase activity.
Mutant strains such as No. 61 (Microtechnical Research Institute No. 3590) can be mentioned. However, it is not limited to these bacteria.
【００１１】突然変異操作としては、放射線（Ｘ線、γ
線、中性子線）や紫外線を照射したり、高熱処理を行っ
たり、また微生物を適当なバッファー中などに懸濁し、
変異源を加えて一定時間インキュベート後、適当に希釈
して寒天培地に植菌し、変異株のコロニーを得るといっ
た操作を行うこともできる。The mutation operation includes radiation (X-ray, γ
Radiation, neutron beam) or ultraviolet light, high heat treatment, or suspending microorganisms in a suitable buffer,
It is also possible to add a mutagen, incubate for a certain period of time, appropriately dilute, inoculate on an agar medium, and obtain a colony of the mutant strain.
【００１２】変異源としては、ナイトロジェンマスター
ド、メチルメタンサルホネート（ＭＭＳ）、Ｎ−メチル
−Ｎ′−ニトロソ−Ｎ−ニトロソグアニジン（ＮＴＧ）
等のアルキル化剤や、５−ブロモウラシル等の塩基類似
体や、マイトマイシンＣ等の抗生物質や、６−メルカプ
トプリン等の塩基合成阻害剤や、プロフラビン等の色素
類や、４−ニトロキノリン−Ｎ−オキシド等のある種の
発癌剤や塩化マンガン、重クロム酸カリウム、亜硝酸、
ヒドラジン、ヒドロキシルアミン、ホルムアルデヒド、
ニトロフラン化合物類などを挙げることができ、使用す
る微生物は、生育菌体（菌糸など）でも良いし、胞子で
も良い。[0012] Mutants include nitrogen mustard, methyl methanesulfonate (MMS), N-methyl-N'-nitroso-N-nitrosoguanidine (NTG).
, A base analog such as 5-bromouracil, an antibiotic such as mitomycin C, a base synthesis inhibitor such as 6-mercaptopurine, a dye such as proflavine, a 4-nitroquinoline Certain carcinogens such as -N-oxide and manganese chloride, potassium dichromate, nitrous acid,
Hydrazine, hydroxylamine, formaldehyde,
Nitrofuran compounds and the like can be mentioned, and the microorganism to be used may be a growing cell (such as a hypha) or a spore.
【００１３】本発明に使用される菌株を培養するために
は、その菌株の胞子、菌子、又は予め培養して得られた
前培養液を、液体培地に接種し培養する。炭素源として
はグルコース、フラクトース、キシロース、サッカロー
ス、マルトース、可溶性デンプン、糖蜜、グリセロー
ル、マンニトール等の一般的に使用されているものが、
いずれも使用できるが、これらに限られるものではな
い。In order to culture the strain used in the present invention, spores, fungi of the strain or a preculture obtained by pre-culturing are inoculated into a liquid medium and cultured. As a carbon source, glucose, fructose, xylose, saccharose, maltose, soluble starch, molasses, glycerol, those commonly used such as mannitol,
Any of them can be used, but is not limited to these.
【００１４】窒素源としてはペプトン、酵母エキス、麦
芽エキス、肉エキス、カザミノ酸、コーンスティプリカ
ー、大豆蛋白等の天然窒素源の他に、尿素等の有機窒素
源、ならびに硝酸ナトリウム、硝酸アンモニウム、硫酸
アンモニウム等の無機窒素源を用いることができる。こ
の他必要に応じリン酸塩、硫酸マグネシウム、硫酸鉄、
硫酸銅等の無機塩及びビタミン等も微量栄養源として使
用できる。As the nitrogen source, in addition to natural nitrogen sources such as peptone, yeast extract, malt extract, meat extract, casamino acid, corn steep liquor, soybean protein, etc., organic nitrogen sources such as urea, sodium nitrate, ammonium nitrate, An inorganic nitrogen source such as ammonium sulfate can be used. In addition, phosphate, magnesium sulfate, iron sulfate,
Inorganic salts such as copper sulfate and vitamins can also be used as trace nutrient sources.
【００１５】これらの培地成分は微生物の成育を害しな
い濃度であれば特に制限しない。実用上一般に、炭素源
は０．１〜３０重量％、好ましくは１〜１０重量％、窒
素源は０．０１〜５重量％、好ましくは０．１〜２重量
％の濃度とするのが良い。培養温度は５〜４０℃、好ま
しくは２０〜３０℃とし、ＥＰＡを製造する際には好ま
しくは１０〜２０℃とする。さらに培地のpHは４〜１
０、好ましくは６〜９として培養を行う。培養は通常２
〜１０日間行う。These medium components are not particularly limited as long as they do not impair the growth of microorganisms. Generally, in practice, the concentration of the carbon source is 0.1 to 30% by weight, preferably 1 to 10% by weight, and the concentration of the nitrogen source is 0.01 to 5% by weight, preferably 0.1 to 2% by weight. . The culture temperature is 5 to 40 ° C, preferably 20 to 30 ° C, and preferably 10 to 20 ° C when producing EPA. Further, the pH of the medium is 4-1.
Culture is performed at 0, preferably 6-9. Culture is usually 2
Perform for 10 days.
【００１６】また、本発明におけるＰＵＦＡの生産を促
進するため、目的とするＰＵＦＡの基質を培地に添加す
ることができる。たとえばω−６系ＰＵＦＡの基質とし
ては、テトラデカン、ヘキサデカン、オクタデカン等の
炭素数１２〜２０の炭化水素、テトラデカン酸、ヘキサ
デカン酸、オクタデカン酸等の炭素数１２〜２０の脂肪
酸、又はその塩（例えばナトリウム塩またはカリウム
塩）、脂肪酸エステル、又は脂肪酸を構成成分として含
む油脂（例えばオリーブ油、大豆油、綿実油、ヤシ油）
等を挙げることができる。In order to promote the production of PUFA in the present invention, a target PUFA substrate can be added to the medium. For example, as a substrate of the ω-6 PUFA, a hydrocarbon having 12 to 20 carbon atoms such as tetradecane, hexadecane and octadecane, a fatty acid having 12 to 20 carbon atoms such as tetradecanoic acid, hexadecanoic acid and octadecanoic acid, or a salt thereof (for example, Fats and oils containing a fatty acid ester or a fatty acid as a component (eg, olive oil, soybean oil, cottonseed oil, coconut oil)
And the like.
【００１７】本発明において、ＰＵＦＡの収量を向上さ
せるため、培養中のある一定期間、好ましくは全培養期
間中に、培養液中のＤＯを５〜２８ppm に制御しつつ培
養する。このような比較的高いＤＯを、微生物に損傷を
与えることなく維持するためには次の２つの方法が有効
である。一つは、培養槽への通気ガスの圧力調整及び培
養槽通気出口の開度調整によって培養槽内全体の圧力を
上げて通常空気を通気する方法であり、この際、培養槽
内の圧力は０．４〜３kg／cm2G、好ましくは０．８〜２
kg／cm2Gにするとよい。In the present invention, in order to improve the yield of PUFA, the cultivation is performed while controlling the DO in the culture solution to 5 to 28 ppm during a certain period of culturing, preferably during the entire culturing period. In order to maintain such a relatively high DO without damaging microorganisms, the following two methods are effective. One is a method of increasing the pressure in the entire culture tank by adjusting the pressure of the ventilation gas to the culture tank and adjusting the opening of the culture tank ventilation outlet to ventilate the normal air. 0.4-3 kg / cm 2 G, preferably 0.8-2
kg / cm 2 G is recommended.
【００１８】また別の方法としては、通常空気に純酸素
又は酸素濃度の高い（酸素濃度が２１％より高い）ガス
を混合するか、あるいは通常空気から窒素等の酸素以外
の成分を一部又は全て除去した酸素富化空気を、培養槽
に通気することにより、所定量の酸素を供給することが
できる。この際、酸素富化空気の酸素濃度は２５〜７７
％、好ましくは２５〜５４％に設定するとよい。またこ
の方法を実施する場合、培養液中のＤＯは５〜２０ppm
に制御しつつ培養することが好ましい。さらにこれらの
方法は単独でも、または併用してあるいは組み合せて行
なうことができる。なおこれらの方法において、培養槽
への通気は通常、０．１vvm （単位vvm：Ｎｌ／ｌ−ｂ
ｒｏｔｈ／min 、以下すべて同様）以上、好ましくは
０．５〜２vvm の範囲で行なう。As another method, pure oxygen or a gas having a high oxygen concentration (oxygen concentration is higher than 21%) is mixed with ordinary air, or a part of components other than oxygen such as nitrogen such as nitrogen is added from ordinary air. A predetermined amount of oxygen can be supplied by passing the oxygen-enriched air from which all has been removed into a culture tank. At this time, the oxygen concentration of the oxygen-enriched air is 25 to 77.
%, Preferably 25 to 54%. When this method is performed, DO in the culture solution is 5 to 20 ppm.
It is preferable to culture while controlling the temperature. Furthermore, these methods can be performed alone, or in combination or in combination. In these methods, the aeration to the culture tank is usually 0.1 vvm (unit vvm: Nl / lb).
roth / min, the same applies hereinafter) or more, preferably in the range of 0.5 to 2 vvm.
【００１９】本発明では、目的のＰＵＦＡを製造するた
めに、必要であれば従来から知られている酵素活性阻害
剤や活性化剤を用いることができる。このようにして培
養して、菌体内に目的とするＰＵＦＡを含有する脂質が
生成蓄積される。この培養菌体から、通常の方法により
目的とするＰＵＦＡの採取を行う。In the present invention, conventionally known enzyme activity inhibitors and activators can be used, if necessary, to produce the target PUFA. By culturing in this manner, the target PUFA-containing lipid is produced and accumulated in the cells. From the cultured cells, a target PUFA is collected by an ordinary method.
【００２０】[0020]
【実施例】次に、実施例により、この発明をさらに具体
的に説明する。実施例１． グルコース２％、酵母エキス１％を含む培地
（pH６．３）１００mLを５００mLエルレンマイヤーフラ
スコに入れ、１２０℃で２０分殺菌した。モルティエレ
ラ・アルピナ（Ｍｏｒｔｉｅｒｅｌｌａ ａｌｐｉｎ
ａ）ＩＦＯ８５６８を１白金耳植菌し、レシプロシェー
カー（１００rpm)により２８℃で４日間振盪培養し、こ
れを前培養液とした。本培養は、前培養と同じ組成の培
地を５０Ｌ容培養槽に２５Ｌ仕込み、１２０℃で２０分
殺菌・冷却後、前培養液５００mlを接種した。Next, the present invention will be described in more detail with reference to Examples.
Will be explained.Embodiment 1 FIG. Medium containing 2% glucose and 1% yeast extract
(PH 6.3) 100 mL to 500 mL Erlenmeyer Fra
It was put in a scoop and sterilized at 120 ° C. for 20 minutes. Mortiere
La Alpina (Mortierella alpin
a) Inoculate one platinum loop of IFO8568 and reciprocate
Incubate with shaking (100 rpm) at 28 ° C for 4 days.
This was used as a preculture. The main culture is a culture with the same composition as the preculture.
25L of the ground is put into a 50L culture tank, and it is 20 minutes at 120 ° C.
After sterilization and cooling, 500 ml of the preculture was inoculated.
【００２１】本発明法では、培養液のＤＯが培養の全期
間を通じてそれぞれ６〜１１ppm 、１０〜１５ppm 、１
４〜１９ppm になるように、通気中の酸素濃度を調整し
た。ちなみに、この場合の酸素濃度は各々約３０％、約
４１％、約５１％であった。コントロールは通常の空気
（酸素濃度２１％）を通気した。なお、いずれの場合も
培養温度２８℃、攪拌回転数２００rpm 、通気量１vvm
、槽内常圧の条件で７日間培養を行なった。また、培
養１〜５日目には、各々１日当たりグルコース１％（対
ブロス）を添加し、必要に応じて消泡剤を添加して培養
した。培養後濾過により菌体を回収し、十分に水洗した
後、１０５℃で２時間静置して乾燥菌体を得た。In the method of the present invention, the DO of the culture solution is 6 to 11 ppm, 10 to 15 ppm, 1
The oxygen concentration during ventilation was adjusted to be 4 to 19 ppm. Incidentally, the oxygen concentrations in this case were about 30%, about 41%, and about 51%, respectively. The control ventilated with normal air (oxygen concentration 21%). In each case, the culture temperature was 28 ° C., the stirring speed was 200 rpm, and the aeration rate was 1 vvm.
The culture was carried out for 7 days under normal pressure in the tank. On the 1st to 5th days of culture, glucose was added at 1% per day (vs. broth), and an antifoaming agent was added as needed, followed by culturing. After the culture, the cells were collected by filtration, washed sufficiently with water, and allowed to stand at 105 ° C. for 2 hours to obtain dried cells.
【００２２】乾燥菌体より、クロロホルム−メタノール
−水の一層系の溶媒を用いるＢｌｉｇｈ＆Ｄｙｅｒの抽
出法によって総脂質を抽出、さらにこれを無水メタノー
ル−塩酸（９５：５）を用いて２０℃にて３時間処理す
る事によってメチルエステル化し、エーテルで抽出して
脂肪酸メチルを得た。得られた脂肪酸メチルをガスクロ
マトグラフィーで分析した。表１にその結果を示す。The total lipids were extracted from the dried cells by the Blight & Dyer extraction method using a chloroform-methanol-water one-layer solvent, and further extracted with anhydrous methanol-hydrochloric acid (95: 5) at 20 ° C. for 3 hours. The mixture was subjected to methyl esterification by treating with time, and extracted with ether to obtain fatty acid methyl. The resulting fatty acid methyl was analyzed by gas chromatography. Table 1 shows the results.
【００２３】 表 １ 酸素富化空気 通常空気 通気ガス酸素濃度（％） 30 41 51 21 DO値 培養液中の最低DO(ppm) 6.6 10.5 14.2 1.1 培養液中の最高DO(ppm) 10.5 14.9 18.0 7.8 培養成績 乾燥菌体（ｇ／Ｌ） 18.9 19.8 19.2 16.6 総脂肪酸（ｇ／Ｌ） 9.17 9.52 9.30 6.55 ARA(ｇ／Ｌ） 3.77 4.01 3.80 2.61 DGLA（ｇ／Ｌ） 0.39 0.38 0.37 0.20 Table 1 Oxygen-enriched air Normal air ventilation gas oxygen concentration (%) 30 41 51 21 DO value Minimum DO (ppm) in culture solution 6.6 10.5 14.2 1.1 Maximum DO (ppm) in culture solution 10.5 14.9 18.0 7.8 Culture result Dry cells (G / L) 18.9 19.8 19.2 16.6 Total fatty acids (g / L) 9.17 9.52 9.30 6.55 ARA (g / L) 3.77 4.01 3.80 2.61 DGLA (g / L) 0.39 0.38 0.37 0.20
【００２４】表１から明らかなようにコントロールに比
べ、ＤＯを６〜１１ppm に制御の場合、ＡＲＡで４４
％、ＤＧＬＡで９５％、ＤＯを１０〜１５ppm に制御の
場合、ＡＲＡで５４％、ＤＧＬＡで９０％、ＤＯを１４
〜１９ppm に制御の場合、ＡＲＡで４６％、ＤＧＬＡで
８５％、各々生産量が増大した。As is clear from Table 1, when the DO is controlled to 6 to 11 ppm, the ARA is 44
%, 95% DGLA and 10-15 ppm DO, 54% ARA, 90% DGLA and 14% DO.
In the case of controlling to １９19 ppm, the production amount increased by 46% for ARA and 85% for DGLA, respectively.
【００２５】実施例２．グルコース２％、酵母エキス１
％を含む培地（pH６．３）を５０Ｌ容培養槽に２５Ｌで
仕込み、１２０℃で４０分殺菌、冷却後、実施例１と同
様に調製したモルティエレラ・エキシグア（Ｍｏｒｔｉ
ｅｒｅｌｌａ ｅｘｉｇｕａ）ＩＦＯ８５７１の前培養
液を５００mL接種した。本発明法では培養液のＤＯが、
培養の全期間を通じて５〜１０ppm 、１１〜１６ppm 、
１７〜２２ppm 、２３〜２８ppmの範囲となるように培B
R>養槽内の圧力を調整した。 Embodiment 2 FIG . Glucose 2%, yeast extract 1
% Was charged in 25L medium (pH 6.3) to a 50L fermentor containing 40 min sterilized at 120 ° C., after cooling, Mortierella exigua, prepared as in Example 1 (Morti
the erella exigua) IFO8571 of pre-culture solution was 500mL inoculated. In the method of the present invention, the DO of the culture solution is
5-10 ppm throughout the culture, 11-16 ppm,
Medium B to be in the range of 17 to 22 ppm and 23 to 28 ppm
R> The pressure in the nutrient tank was adjusted.
【００２６】ちなみに、この場合の内圧は各々約０．５
kg／cm2G、約１．２kg／cm2G、約２．０kg／cm2G、約
２．６kg／cm2Gであった。コントロールは常圧で培養し
た。なお何れの場合も、培養温度２８℃、攪拌回転数２
００rpm 、通常空気１vvm 通気の条件で７日間の培養を
行った。また、実施例１と同様に、培養１〜５日目のグ
ルコース添加、必要に応じた消泡剤添加を行なった。培
養後、実施例１と同様の方法で菌体回収および脂肪酸メ
チルの調製を行い、ガスクロマトグラフィーで分析し
た。表２にその結果を示す。The internal pressure in this case is about 0.5
kg / cm 2 G, about 1.2 kg / cm 2 G, about 2.0 kg / cm 2 G, and about 2.6 kg / cm 2 G. Controls were cultured at normal pressure. In each case, the culture temperature was 28 ° C., and the stirring rotation speed was 2
Cultivation was performed for 7 days under the conditions of 00 rpm and aeration of 1 vvm with normal air. In the same manner as in Example 1, glucose was added on days 1 to 5 of culture, and an antifoaming agent was added as needed. After the culture, the cells were collected and fatty acid methyl was prepared in the same manner as in Example 1, and analyzed by gas chromatography. Table 2 shows the results.
【００２７】 表 ２ 本 発 明 法 コントロール 培養槽内圧力（kg／cm2G) 0.5 1.2 2.0 2.6 0 培養中の最低DO(ppm) 5.0 12.5 17.0 23.2 1.1 培養中の最高DO(ppm) 9.8 15.4 21.3 27.5 7.8 乾燥菌体（ｇ／Ｌ） 18.9 20.7 21.1 17.5 16.2 総脂肪酸（ｇ／Ｌ） 7.46 8.20 8.25 7.01 6.56 ARA(ｇ／Ｌ） 3.06 3.63 3.72 2.86 2.55 DGLA（ｇ／Ｌ） 0.32 0.41 0.43 0.31 0.24 Table 2 Inventive method Pressure in control culture tank (kg / cm 2 G) 0.5 1.2 2.0 2.6 0 Lowest DO (ppm) during culture 5.0 12.5 17.0 23.2 1.1 Highest DO (ppm) during culture 9.8 15.4 21.3 27.5 7.8 Dry cells (G / L) 18.9 20.7 21.1 17.5 16.2 Total fatty acids (g / L) 7.46 8.20 8.25 7.01 6.56 ARA (g / L) 3.06 3.63 3.72 2.86 2.55 DGLA (g / L) 0.32 0.41 0.43 0.31 0.24
【００２８】表２から明らかなようにコントロールに比
べ、ＤＯを５〜１０ppm に制御の場合、ＡＲＡで２０
％、ＤＧＬＡで３３％、ＤＯを１１〜１６ppm に制御の
場合、ＡＲＡで４２％、ＤＧＬＡで７１％、ＤＯを１７
〜２２ppm に制御の場合、ＡＲＡで４６％、ＤＧＬＡで
７９％、ＤＯを２３〜２８ppm に制御の場合、ＡＲＡで
１２％、ＤＧＬＡで２９％、各々生産量が増大した。As is clear from Table 2, when DO is controlled to 5 to 10 ppm, ARA is 20
%, DGLA at 33%, DO at 11-16 ppm, ARA at 42%, DGLA at 71%, DO at 17%.
When controlling to ~ 22 ppm, ARA increased 46%, DGLA 79%, and when controlling DO to 23-28 ppm, ARA increased 12% and DGLA increased 29%, respectively.
【００２９】実施例３．グルコース０．５％、オリーブ
油２％、酵母エキス１％を含む培地（pH６．０）を５０
Ｌ容培養槽に２５Ｌ仕込み、１２０℃で２０分殺菌、冷
却後、実施例１と同様に調製したモルティエレラ・エロ
ンガタ（Ｍｏｒｔｉｅｒｅｌｌａ ｅｌｏｎｇａｔａ）
ＩＦＯ８５７０の前培養液５００mlを接種した。本実施
例では、培養液のＤＯが培養の全期間を通じて１１〜１
６ppm 、２５〜３０ppm 、３０〜３５ppm になるよう
に、通気中の酸素濃度を調整した。ちなみに、この場合
の酸素濃度は各々約４３％、約８１％、約９５％であっ
た。コントロールは通常の空気（酸素濃度２１％）を通
気した。 Embodiment 3 FIG . A medium (pH 6.0) containing 0.5% glucose, 2% olive oil, and 1% yeast extract was added to 50
After adding 25 L to an L-volume culture tank, sterilizing at 120 ° C. for 20 minutes, and cooling, Mortierella elongata prepared as in Example 1 was used.
500 ml of a preculture of IFO8570 was inoculated. In this example, the DO of the culture solution was 11 to 1 throughout the culture.
The oxygen concentration during ventilation was adjusted to 6 ppm, 25 to 30 ppm, and 30 to 35 ppm. Incidentally, the oxygen concentration in this case was about 43%, about 81%, and about 95%, respectively. The control ventilated with normal air (oxygen concentration 21%).
【００３０】なお、いずれの場合も培養温度２８℃、攪
拌回転数２００rpm 、通気量１vvm、槽内常圧の条件で
７日間培養を行なった。また、培養１〜５日目には、各
々１日当たりグルコース１％（対ブロス）を添加し、必
要に応じて消泡剤を添加して培養した。培養後、実施例
１と同様の方法で菌体回収および脂肪酸メチルの調製を
行い、ガスクロマトグラフィーで分析した。表３にその
結果を示す。In each case, cultivation was carried out for 7 days at a culture temperature of 28 ° C., a stirring rotation speed of 200 rpm, an aeration rate of 1 vvm, and a normal pressure in the tank. On the 1st to 5th days of culture, glucose was added at 1% per day (vs. broth), and an antifoaming agent was added as needed, followed by culturing. After the culture, the cells were collected and fatty acid methyl was prepared in the same manner as in Example 1, and analyzed by gas chromatography. Table 3 shows the results.
【００３１】 表 ３ 酸素富化空気通気 通常空気通気 通気ガス酸素濃度（％） 43 81 95 21 DO値 培養液中の最低DO(ppm) 11.0 25.4 31.0 1.4 培養液中の最高DO(ppm) 15.3 29.9 34.4 7.6 培養成績 乾燥菌体（ｇ／Ｌ） 19.8 15.6 12.3 16.7 総脂肪酸（ｇ／Ｌ） 9.52 7.49 5.88 7.98 ARA(ｇ／Ｌ） 3.83 2.85 2.00 3.03 DGLA（ｇ／Ｌ） 0.42 0.33 0.25 0.36 Table 3 Oxygen-enriched air aeration Normal air aeration aeration gas Oxygen concentration (%) 43 81 95 21 DO value Minimum DO (ppm) in culture 11.0 25.4 31.0 1.4 Maximum DO (ppm) in culture 15.3 29.9 34.4 7.6 Culture result Drying Bacteria (g / L) 19.8 15.6 12.3 16.7 Total fatty acids (g / L) 9.52 7.49 5.88 7.98 ARA (g / L) 3.83 2.85 2.00 3.03 DGLA (g / L) 0.42 0.33 0.25 0.36
【００３２】表３から明らかなように、本発明法でＤＯ
を１１〜１６ppm に制御した場合、コントロールに比べ
ＡＲＡで２６％、ＤＧＬＡで１７％生産量が増大した。
しかし、ＤＯを２５〜３０ppm に制御した場合はＡＲＡ
で６％、ＤＧＬＡで８％の生産量減少、さらに、ＤＯを
３０〜３５ppm に制御した場合はＡＲＡで３４％、ＤＧ
ＬＡで３１％の生産量減少が起こった。As is evident from Table 3, DO according to the method of the present invention.
Was controlled to 11 to 16 ppm, the production increased by 26% in ARA and 17% in DGLA as compared with the control.
However, when DO is controlled to 25 to 30 ppm, ARA
6% in DGLA, 8% in DGLA, and 34% in ARA and DG when DO is controlled to 30-35 ppm.
LA resulted in a 31% reduction in production.
【００３３】実施例４．グルコース０．５％、オリーブ
油２％、酵母エキス１％を含む培地（pH６．０）を５０
Ｌ容培養槽に２５Ｌで仕込み、１２０℃で４０分殺菌、
冷却後、実施例１と同様に調製したモルティエレラ・ヒ
グロフィラ（Ｍｏｒｔｉｅｒｅｌｌａｈｙｇｒｏｐｈｉ
ｌａ）ＩＦＯ５９４１の前培養液を５００mL接種した。
本実施例では培養液のＤＯが、培養の全期間を通じて９
〜１３ppm 、２８〜３３ppm の範囲となるように培養槽
内の圧力を調整した。 Embodiment 4 FIG . A medium (pH 6.0) containing 0.5% glucose, 2% olive oil, and 1% yeast extract was added to 50
25 L into an L-culture tank, sterilized at 120 ° C for 40 minutes,
After cooling, Mortierella hygrophila prepared as in Example 1
la ) 500 mL of the preculture of IFO5941 was inoculated.
In this example, the DO of the culture solution was 9 during the entire period of the culture.
The pressure in the culture tank was adjusted so as to be in the range of １３13 ppm and 28-33 ppm.
【００３４】ちなみに、この場合の内圧は約０．９kg／
cm2G、約３．２kg／cm2Gであった。コントロールは常圧
で培養した。なお何れの場合も、培養温度２８℃、攪拌
回転数２００rpm 、通常空気１vvm 通気の条件で７日間
培養を行なった。また、実施例１と同様に、培養１〜５
日目のグルコース添加、必要に応じた消泡剤添加を行な
った。培養後、実施例１と同様の方法で菌体回収および
脂肪酸メチルの調製を行い、ガスクロマトグラフィーで
分析した。表４にその結果を示す。Incidentally, the internal pressure in this case is about 0.9 kg /
cm 2 G, about 3.2 kg / cm 2 G. Controls were cultured at normal pressure. In each case, the culture was carried out for 7 days under the conditions of a culture temperature of 28 ° C., a stirring rotation speed of 200 rpm and a normal air flow of 1 vvm. In addition, as in Example 1, cultures 1 to 5
On the day, glucose was added, and if necessary, an antifoaming agent was added. After the culture, the cells were collected and fatty acid methyl was prepared in the same manner as in Example 1, and analyzed by gas chromatography. Table 4 shows the results.
【００３５】 表 ４ 槽内加圧 槽内常圧 培養槽内圧力（kg／cm2G) 0.9 3.2 0 培養中の最低DO(ppm) 9.2 28.5 1.4 培養中の最高DO(ppm) 12.9 32.2 7.5 乾燥菌体（ｇ／Ｌ） 22.3 16.4 17.2 総脂肪酸（ｇ／Ｌ） 10.9 8.02 8.41 ARA(ｇ／Ｌ） 3.92 2.49 2.78 DGLA（ｇ／Ｌ） 0.90 0.67 0.69 Table 4 Tank pressurization Tank normal pressure culture tank pressure (kg / cm 2 G) 0.9 3.2 0 Lowest DO (ppm) during culture 9.2 28.5 1.4 Highest DO (ppm) during culture 12.9 32.2 7.5 Dry cells (g / cm2 L) 22.3 16.4 17.2 Total fatty acids (g / L) 10.9 8.02 8.41 ARA (g / L) 3.92 2.49 2.78 DGLA (g / L) 0.90 0.67 0.69
【００３６】表４から明らかなように、本発明法でＤＯ
を９〜１３ppm に制御した場合、コントロールに比べＡ
ＲＡで４１％、ＤＧＬＡで３０％生産量が増大した。し
かし、ＤＯを２８〜３３ppm に制御した場合はＡＲＡで
１０％、ＤＧＬＡで３％の生産量減少が起こった。As is apparent from Table 4, DO according to the method of the present invention was used.
Is controlled to 9 to 13 ppm, A
RA increased by 41% and DGLA by 30%. However, when the DO was controlled at 28 to 33 ppm, the production decreased by 10% in ARA and 3% in DGLA.
【００３７】実施例５．グルコース１．８％、酵母エキ
ス１％、オリブ油０．５％を含む培地（pH６．３）を５
０Ｌ容培養槽に２５Ｌで仕込み、１２０℃で４０分殺
菌、冷却後、実施例１と同様に調製したモルティエレラ
・エロンガタ（Ｍｏｒｔｉｅｒｅｌｌａｅｌｏｎｇａｔ
ａ）ＳＡＭ１８６０の前培養液を５００mL接種した。本
発明法では培養液のＤＯが、培養の全期間を通じて１２
〜１７ppm の範囲となるように、酸素富化空気と培養槽
内の圧力で調整した。 Embodiment 5 FIG . A medium (pH 6.3) containing 1.8% glucose, 1% yeast extract, and 0.5% olive oil was added to 5
After charging 25 L into a 0 L culture tank, sterilizing at 120 ° C. for 40 minutes, and cooling, Mortierella elongata prepared in the same manner as in Example 1 was used.
a ) 500 mL of SAM1860 preculture was inoculated. In the method of the present invention, the DO of the culture solution is 12
The concentration was adjusted with oxygen-enriched air and the pressure in the culture tank so as to be in the range of １７17 ppm.
【００３８】ちなみに、この場合の酸素濃度は約２５
％、内圧は約０．８kg／cm2Gであった。コントロールは
通常の空気を常圧で通気した。なお何れの場合も、培養
温度２８℃、攪拌回転数２００rpm 、通気量１vvm 通気
の条件で７日間培養を行なった。また、実施例１と同様
に、培養１〜５日目のグルコース添加、必要に応じた消
泡剤添加を行なった。培養後、実施例１と同様の方法で
菌体回収および脂肪酸メチルの調製を行い、ガスクロマ
トグラフィーで分析した。表５にその結果を示す。By the way, the oxygen concentration in this case is about 25
%, The internal pressure was about 0.8 kg / cm 2 G. Controls were ventilated with normal air at normal pressure. In each case, the culture was carried out for 7 days under the conditions of a culture temperature of 28 ° C., a stirring rotation speed of 200 rpm, and an aeration of 1 vvm. In the same manner as in Example 1, glucose was added on days 1 to 5 of culture, and an antifoaming agent was added as needed. After the culture, the cells were collected and fatty acid methyl was prepared in the same manner as in Example 1, and analyzed by gas chromatography. Table 5 shows the results.
【００３９】 表 ５ Ｉ（本発明法） II（コントロール） 培養中の最低DO(ppm) 13.1 1.6 培養中の最高DO(ppm) 16.6 7.7 乾燥菌体（ｇ／Ｌ） 22.3 17.2 総脂肪酸（ｇ／Ｌ） 10.2 8.03 DGLA（ｇ／Ｌ） 3.72 2.94 ARA(ｇ／Ｌ） 0.95 0.80 表５から明らかなように、本発明法の場合、コントロー
ルに比べてＤＧＬＡで２７％、ＡＲＡで１９％生産量が
増大した。 Table 5 I (the method of the present invention) II (control) Lowest DO (ppm) in culture 13.1 1.6 Highest DO (ppm) in culture 16.6 7.7 Dried cells (g / L) 22.3 17.2 Total fatty acids (g / L) 10.2 8.03 DGLA (G / L) 3.72 2.94 ARA (g / L) 0.95 0.80 As is clear from Table 5, in the case of the method of the present invention, DGLA and ARA increased the production by 27% and 19%, respectively, as compared with the control.
【００４０】実施例６．グルコース０．５％、オリーブ
油２％、酵母エキス１％を含む培地（pH６．０）を５０
Ｌ容培養槽に２５Ｌ仕込み、１２０℃で２０分殺菌、冷
却後、実施例１と同様に調製したモルティエレラ・エロ
ンガタ（Ｍｏｒｔｉｅｒｅｌｌａ ｅｌｏｎｇａｔａ）
ＳＡＭ０２１９の前培養液５００mlを接種した。本発明
法では、培養液のＤＯが培養の全期間を通じて２２〜２
７ppm になるように、通気中の酸素濃度と培養槽内圧力
で調整した。 Embodiment 6 FIG . A medium (pH 6.0) containing 0.5% glucose, 2% olive oil, and 1% yeast extract was added to 50
After adding 25 L to an L-volume culture tank, sterilizing at 120 ° C. for 20 minutes, and cooling, Mortierella elongata prepared as in Example 1 was used.
500 ml of a preculture of SAM0219 was inoculated. In the method of the present invention, the DO of the culture solution is 22 to 2 throughout the culture.
The oxygen concentration during aeration and the pressure in the culture tank were adjusted to 7 ppm.
【００４１】ちなみに、この場合の酸素濃度は約２９
％、内圧は約１．５kg／cm2Gであった。コントロールは
通常の空気（酸素濃度２１％）を常圧で通気した。な
お、いずれの場合も培養温度２８℃、攪拌回転数２００
rpm 、１vvm 通気の条件で７日間培養を行なった。ま
た、培養１〜５日目には、各々１日当たりグルコース１
％（対ブロス）を添加し、必要に応じて消泡剤を添加し
て培養した。培養後、実施例１と同様の方法で菌体回収
および脂肪酸メチルの調製を行い、ガスクロマトグラフ
ィーで分析した。表６にその結果を示す。Incidentally, the oxygen concentration in this case is about 29
%, And the internal pressure was about 1.5 kg / cm 2 G. For control, normal air (oxygen concentration 21%) was ventilated at normal pressure. In each case, the culture temperature was 28 ° C. and the number of rotations was 200.
Culturing was performed for 7 days under the conditions of rpm and 1 vvm aeration. On the 1st to 5th days of culture, glucose 1
% (Vs. broth) and, if necessary, an antifoaming agent. After the culture, the cells were collected and fatty acid methyl was prepared in the same manner as in Example 1, and analyzed by gas chromatography. Table 6 shows the results.
【００４２】 表 ６ Ｉ（本発明法） II（コントロール） DO値 培養液中の最低DO(ppm) 22.5 1.4 培養液中の最高DO(ppm) 26.2 7.6 培養成績 乾燥菌体（ｇ／Ｌ） 21.4 17.3 総脂肪酸（ｇ／Ｌ） 10.5 8.13 ARA(ｇ／Ｌ） 4.09 3.09 DGLA（ｇ／Ｌ） 0.63 0.41 表６から明らかなように、本発明法の場合、コントロー
ルに比べＡＲＡで３２％、ＤＧＬＡで５４％生産量が増
大した。 Table 6 I (the method of the present invention) II (control) DO value Lowest DO (ppm) in culture solution 22.5 1.4 Highest DO (ppm) in culture solution 26.2 7.6 Culture results Dried cells (g / L) 21.4 17.3 Total fatty acids (g) / L) 10.5 8.13 ARA (g / L) 4.09 3.09 DGLA (g / L) 0.63 0.41 As is evident from Table 6, in the case of the method of the present invention, the production was increased by 32% in ARA and 54% in DGLA as compared with the control.
───────────────────────────────────────────────────── フロントページの続き (72)発明者 新免 芳史 東京都府中市緑町２−18−９ アルト東 府中302 (72)発明者 松元 信也 大阪府三島郡島本町桜井台８−15 (72)発明者 山田 秀明 京都府京都市左京区松ヶ崎木ノ本町19− １ (56)参考文献 特開 平２−142486（ＪＰ，Ａ) 特開 昭60−168391（ＪＰ，Ａ) 特開 昭63−44891（ＪＰ，Ａ) 特開 昭63−14697（ＪＰ，Ａ) 特開 平３−272692（ＪＰ，Ａ) 特開 平１−243992（ＪＰ，Ａ) 特開 平２−23878（ＪＰ，Ａ) 特開 平３−19696（ＪＰ，Ａ) 特開 平５−302087（ＪＰ，Ａ) 米国特許5166366（ＵＳ，Ａ) 国際公開91／11918（ＷＯ，Ａ１) 日本食品工業学会誌（1980），Ｖｏ ｌ．27，Ｎｏ．１，ｐ．20−24 カナダ国特許出願公開2070697号明細 書 (58)調査した分野(Int.Cl.7，ＤＢ名) C12P 3/00 - 11/00 ＢＩＯＳＩＳ（ＤＩＡＬＯＧ) ＷＰＩ（ＤＩＡＬＯＧ) ＪＩＣＳＴファイル（ＪＯＩＳ)──────────────────────────────────────────────────続 き Continued on the front page (72) Inventor Yoshifumi Shinmen 2-18-9 Midoricho, Fuchu-shi, Tokyo Alto-Higashi Fuchu 302 (72) Inventor Shinya Matsumoto 8-15 Sakuraidai, Shimamotocho, Mishima-gun, Osaka (72) Inventor Hideaki Yamada 19-1 Matsugasaki Kinohonmachi, Sakyo-ku, Kyoto-shi, Kyoto (56) Reference JP-A-2-142486 (JP, A) JP-A-60-168391 (JP, A) JP-A-63-44891 ( JP, A) JP-A-63-14697 (JP, A) JP-A-3-272692 (JP, A) JP-A 1-224392 (JP, A) JP-A-2-23878 (JP, A) JP JP-A-3-19696 (JP, A) JP-A-5-302087 (JP, A) U.S. Pat. No. 5,166,366 (US, A) International Publication No. 91/11918 (WO, A1) Journal of the Japan Food Industry Association (1980), Vol. 27, No. 1, p. 20-24 Canadian Patent Application Publication No. 2070697 (58) Fields investigated (Int. Cl. 7 , DB name) C12P 3/00-11/00 BIOSIS (DIALOG) WPI (DIALOG) JICST file (JOIS)
Claims (5)
ティエレラ(Mortierella）亜属に属する微生物を液体培
地中で通気培養して、高度不飽和脂肪酸及びこれを含有
する脂質を製造する方法において、培養液中の溶存酸素
濃度を9.2〜28ppm に維持することを特徴とする、高度
不飽和脂肪酸又はこれを含有する脂質の製造方法。1. A method for producing a polyunsaturated fatty acid and a lipid containing the same by subjecting a microorganism belonging to the subgenus Mortierella of the genus Mortierella to aeration culture in a liquid medium. A method for producing a polyunsaturated fatty acid or a lipid containing the same, characterized in that the concentration of dissolved oxygen therein is maintained at 9.2 to 28 ppm.
状態に維持することを特徴とする、請求項１記載の高度
不飽和脂肪酸又はこれを含有する脂質の製造方法。2. The method for producing a polyunsaturated fatty acid or a lipid containing the same according to claim 1, further comprising maintaining the pressure in the culture tank during the culturing in a pressurized state.
素富化空気を、通気することを特徴とする、請求項１記
載の高度不飽和脂肪酸又はこれを含有する脂質の製造方
法。3. The method for producing a polyunsaturated fatty acid or a lipid containing the same according to claim 1, further comprising aerating oxygen-enriched air having an oxygen concentration higher than ordinary air.
に維持することを特徴とする請求項３記載の高度不飽和
脂肪酸又はこれを含有する脂質の製造方法。4. The concentration of dissolved oxygen in a culture solution is 9.2 to 20 ppm.
The method for producing a highly unsaturated fatty acid or a lipid containing the same according to claim 3, wherein
〜４のいずれか１項に記載の高度不飽和脂肪酸又はこれ
を含有する脂質の製造方法。 5. The method according to claim 1, wherein the culture temperature is 20 ° C. to 30 ° C.
Polyunsaturated fatty acid according to any one of to 4 or 4, or
A method for producing a lipid comprising:
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|---|---|---|---|
| JP30552392A JP3354608B2 (en) | 1992-11-16 | 1992-11-16 | Method for producing polyunsaturated fatty acid and lipid containing the same |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP30552392A JP3354608B2 (en) | 1992-11-16 | 1992-11-16 | Method for producing polyunsaturated fatty acid and lipid containing the same |
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| Publication Number | Publication Date |
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| JPH06153970A JPH06153970A (en) | 1994-06-03 |
| JP3354608B2 true JP3354608B2 (en) | 2002-12-09 |
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| JP30552392A Expired - Lifetime JP3354608B2 (en) | 1992-11-16 | 1992-11-16 | Method for producing polyunsaturated fatty acid and lipid containing the same |
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| JP (1) | JP3354608B2 (en) |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| US5658767A (en) * | 1991-01-24 | 1997-08-19 | Martek Corporation | Arachidonic acid and methods for the production and use thereof |
| PT960943E (en) | 1996-12-27 | 2014-04-29 | Suntory Holdings Ltd | Media for culturing microorganisms and process for producing unsaturated fatty acids or lipids containing the same |
| JP4127860B2 (en) * | 1997-03-04 | 2008-07-30 | サントリー株式会社 | Polyunsaturated fatty acid and method for producing lipid containing the same |
| DK1122304T3 (en) | 1999-08-13 | 2011-11-21 | Suntory Holdings Ltd | Microorganism secreting a lipid and method for preparing the lipid and lipid balls with lipid encapsulated therein using the microorganism |
| JP4088097B2 (en) | 2002-04-26 | 2008-05-21 | サントリー株式会社 | Method for producing highly unsaturated fatty acid-containing lipid |
| MXPA04012913A (en) | 2002-06-19 | 2005-03-31 | Dsm Ip Assets Bv | Pasteurisation process for microbial cells and microbial oil. |
| DK2966172T3 (en) | 2002-10-11 | 2018-04-23 | Nippon Suisan Kaisha Ltd | PROCEDURE FOR MANUFACTURING MICROBIAL FAT OR OIL WITH LOWER INCOMPATIBLE INGREDIENTS AND SAME FAT OR OIL |
| WO2006016702A1 (en) | 2004-08-12 | 2006-02-16 | Suntory Limited | Method for polyunsaturated fatty acid production using novel cell preservation technique |
| JP4624040B2 (en) * | 2004-09-06 | 2011-02-02 | 日本水産株式会社 | Cell culture method |
| US8241868B2 (en) | 2005-02-08 | 2012-08-14 | Nippon Suisan Kaisha, Ltd. | Production of polyunsaturated fatty acids using cell treatment method |
| JP4849806B2 (en) | 2005-02-08 | 2012-01-11 | 日本水産株式会社 | Method for producing polyunsaturated fatty acids using novel cell treatment method |
| JP6544026B2 (en) * | 2015-04-30 | 2019-07-17 | 株式会社Ihi | Gas supply device of microorganism multiplication equipment |
| WO2017110970A1 (en) * | 2015-12-24 | 2017-06-29 | 花王株式会社 | Method for producing organic acid |
-
1992
- 1992-11-16 JP JP30552392A patent/JP3354608B2/en not_active Expired - Lifetime
Non-Patent Citations (2)
| Title |
|---|
| カナダ国特許出願公開２０７０６９７号明細書 |
| 日本食品工業学会誌（１９８０），Ｖｏｌ．２７，Ｎｏ．１，ｐ．２０−２４ |
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| Publication number | Publication date |
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