JPH05123176A - Production of highly unsaturated fatty acid phospholipid - Google Patents
Production of highly unsaturated fatty acid phospholipidInfo
- Publication number
- JPH05123176A JPH05123176A JP32967390A JP32967390A JPH05123176A JP H05123176 A JPH05123176 A JP H05123176A JP 32967390 A JP32967390 A JP 32967390A JP 32967390 A JP32967390 A JP 32967390A JP H05123176 A JPH05123176 A JP H05123176A
- Authority
- JP
- Japan
- Prior art keywords
- scrc
- genus
- microorganism
- fatty acid
- unsaturated fatty
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 235000021122 unsaturated fatty acids Nutrition 0.000 title claims abstract description 18
- 238000004519 manufacturing process Methods 0.000 title claims description 11
- -1 unsaturated fatty acid phospholipid Chemical class 0.000 title abstract description 9
- 244000005700 microbiome Species 0.000 claims abstract description 44
- 150000004670 unsaturated fatty acids Chemical class 0.000 claims abstract description 12
- 241000589516 Pseudomonas Species 0.000 claims abstract description 9
- 241000588722 Escherichia Species 0.000 claims abstract description 6
- 238000012258 culturing Methods 0.000 claims abstract description 6
- 241000590031 Alteromonas Species 0.000 claims abstract description 5
- 241000607720 Serratia Species 0.000 claims abstract description 4
- 241000863430 Shewanella Species 0.000 claims abstract description 3
- 150000003904 phospholipids Chemical class 0.000 claims description 35
- 238000000034 method Methods 0.000 claims description 11
- 241000863432 Shewanella putrefaciens Species 0.000 claims description 9
- 235000020777 polyunsaturated fatty acids Nutrition 0.000 claims description 9
- JZNWSCPGTDBMEW-UHFFFAOYSA-N Glycerophosphorylethanolamin Natural products NCCOP(O)(=O)OCC(O)CO JZNWSCPGTDBMEW-UHFFFAOYSA-N 0.000 claims description 6
- ATBOMIWRCZXYSZ-XZBBILGWSA-N [1-[2,3-dihydroxypropoxy(hydroxy)phosphoryl]oxy-3-hexadecanoyloxypropan-2-yl] (9e,12e)-octadeca-9,12-dienoate Chemical compound CCCCCCCCCCCCCCCC(=O)OCC(COP(O)(=O)OCC(O)CO)OC(=O)CCCCCCC\C=C\C\C=C\CCCCC ATBOMIWRCZXYSZ-XZBBILGWSA-N 0.000 claims description 5
- AWUCVROLDVIAJX-UHFFFAOYSA-N alpha-glycerophosphate Natural products OCC(O)COP(O)(O)=O AWUCVROLDVIAJX-UHFFFAOYSA-N 0.000 claims description 5
- 150000008104 phosphatidylethanolamines Chemical class 0.000 claims description 5
- 241000193830 Bacillus <bacterium> Species 0.000 claims description 4
- 241000607715 Serratia marcescens Species 0.000 claims description 4
- 241000191940 Staphylococcus Species 0.000 claims description 4
- 241000193755 Bacillus cereus Species 0.000 claims description 3
- 244000063299 Bacillus subtilis Species 0.000 claims description 3
- 235000014469 Bacillus subtilis Nutrition 0.000 claims description 3
- 241000191967 Staphylococcus aureus Species 0.000 claims description 3
- ZGSPNIOCEDOHGS-UHFFFAOYSA-L disodium [3-[2,3-di(octadeca-9,12-dienoyloxy)propoxy-oxidophosphoryl]oxy-2-hydroxypropyl] 2,3-di(octadeca-9,12-dienoyloxy)propyl phosphate Chemical compound [Na+].[Na+].CCCCCC=CCC=CCCCCCCCC(=O)OCC(OC(=O)CCCCCCCC=CCC=CCCCCC)COP([O-])(=O)OCC(O)COP([O-])(=O)OCC(OC(=O)CCCCCCCC=CCC=CCCCCC)COC(=O)CCCCCCCC=CCC=CCCCCC ZGSPNIOCEDOHGS-UHFFFAOYSA-L 0.000 claims description 2
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 abstract description 27
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 abstract description 24
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 abstract description 23
- MBMBGCFOFBJSGT-KUBAVDMBSA-N all-cis-docosa-4,7,10,13,16,19-hexaenoic acid Chemical compound CC\C=C/C\C=C/C\C=C/C\C=C/C\C=C/C\C=C/CCC(O)=O MBMBGCFOFBJSGT-KUBAVDMBSA-N 0.000 abstract description 23
- 239000000203 mixture Substances 0.000 abstract description 15
- 235000020669 docosahexaenoic acid Nutrition 0.000 abstract description 13
- 235000019197 fats Nutrition 0.000 abstract description 13
- 235000020673 eicosapentaenoic acid Nutrition 0.000 abstract description 12
- 150000002632 lipids Chemical class 0.000 abstract description 11
- 239000000843 powder Substances 0.000 abstract description 11
- 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 abstract description 10
- 229940090949 docosahexaenoic acid Drugs 0.000 abstract description 10
- JAZBEHYOTPTENJ-UHFFFAOYSA-N eicosapentaenoic acid Natural products CCC=CCC=CCC=CCC=CCC=CCCCC(O)=O JAZBEHYOTPTENJ-UHFFFAOYSA-N 0.000 abstract description 10
- 229960005135 eicosapentaenoic acid Drugs 0.000 abstract description 10
- 230000000813 microbial effect Effects 0.000 abstract description 9
- 239000002904 solvent Substances 0.000 abstract description 7
- 241000588724 Escherichia coli Species 0.000 abstract description 4
- 238000010898 silica gel chromatography Methods 0.000 abstract description 4
- 239000002537 cosmetic Substances 0.000 abstract description 3
- 239000003814 drug Substances 0.000 abstract description 3
- 235000013305 food Nutrition 0.000 abstract description 3
- 238000000926 separation method Methods 0.000 abstract description 3
- 238000000605 extraction Methods 0.000 abstract description 2
- 150000001875 compounds Chemical class 0.000 abstract 2
- 210000004748 cultured cell Anatomy 0.000 abstract 1
- 238000001035 drying Methods 0.000 abstract 1
- 238000005406 washing Methods 0.000 abstract 1
- 239000003925 fat Substances 0.000 description 13
- 239000000470 constituent Substances 0.000 description 12
- 210000004027 cell Anatomy 0.000 description 10
- 235000014113 dietary fatty acids Nutrition 0.000 description 10
- 229930195729 fatty acid Natural products 0.000 description 10
- 239000000194 fatty acid Substances 0.000 description 10
- 239000003921 oil Substances 0.000 description 9
- 235000019198 oils Nutrition 0.000 description 9
- 150000004665 fatty acids Chemical class 0.000 description 8
- 102220201851 rs143406017 Human genes 0.000 description 7
- AZQWKYJCGOJGHM-UHFFFAOYSA-N 1,4-benzoquinone Chemical compound O=C1C=CC(=O)C=C1 AZQWKYJCGOJGHM-UHFFFAOYSA-N 0.000 description 6
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 6
- 238000004809 thin layer chromatography Methods 0.000 description 6
- 235000014593 oils and fats Nutrition 0.000 description 5
- LRHPLDYGYMQRHN-UHFFFAOYSA-N N-Butanol Chemical compound CCCCO LRHPLDYGYMQRHN-UHFFFAOYSA-N 0.000 description 4
- IPCSVZSSVZVIGE-UHFFFAOYSA-N hexadecanoic acid Chemical compound CCCCCCCCCCCCCCCC(O)=O IPCSVZSSVZVIGE-UHFFFAOYSA-N 0.000 description 4
- SECPZKHBENQXJG-FPLPWBNLSA-N palmitoleic acid Chemical compound CCCCCC\C=C/CCCCCCCC(O)=O SECPZKHBENQXJG-FPLPWBNLSA-N 0.000 description 4
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 3
- 229930186217 Glycolipid Natural products 0.000 description 3
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 3
- 238000005119 centrifugation Methods 0.000 description 3
- 238000001816 cooling Methods 0.000 description 3
- 239000012046 mixed solvent Substances 0.000 description 3
- 239000003960 organic solvent Substances 0.000 description 3
- WRIDQFICGBMAFQ-UHFFFAOYSA-N (E)-8-Octadecenoic acid Natural products CCCCCCCCCC=CCCCCCCC(O)=O WRIDQFICGBMAFQ-UHFFFAOYSA-N 0.000 description 2
- TWJNQYPJQDRXPH-UHFFFAOYSA-N 2-cyanobenzohydrazide Chemical compound NNC(=O)C1=CC=CC=C1C#N TWJNQYPJQDRXPH-UHFFFAOYSA-N 0.000 description 2
- LQJBNNIYVWPHFW-UHFFFAOYSA-N 20:1omega9c fatty acid Natural products CCCCCCCCCCC=CCCCCCCCC(O)=O LQJBNNIYVWPHFW-UHFFFAOYSA-N 0.000 description 2
- QSBYPNXLFMSGKH-UHFFFAOYSA-N 9-Heptadecensaeure Natural products CCCCCCCC=CCCCCCCCC(O)=O QSBYPNXLFMSGKH-UHFFFAOYSA-N 0.000 description 2
- 241000251468 Actinopterygii Species 0.000 description 2
- 241000555825 Clupeidae Species 0.000 description 2
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 2
- 241000196324 Embryophyta Species 0.000 description 2
- 241001465754 Metazoa Species 0.000 description 2
- 235000021360 Myristic acid Nutrition 0.000 description 2
- TUNFSRHWOTWDNC-UHFFFAOYSA-N Myristic acid Natural products CCCCCCCCCCCCCC(O)=O TUNFSRHWOTWDNC-UHFFFAOYSA-N 0.000 description 2
- 239000005642 Oleic acid Substances 0.000 description 2
- ZQPPMHVWECSIRJ-UHFFFAOYSA-N Oleic acid Natural products CCCCCCCCC=CCCCCCCCC(O)=O ZQPPMHVWECSIRJ-UHFFFAOYSA-N 0.000 description 2
- 235000021314 Palmitic acid Nutrition 0.000 description 2
- 235000021319 Palmitoleic acid Nutrition 0.000 description 2
- 239000001888 Peptone Substances 0.000 description 2
- 108010080698 Peptones Proteins 0.000 description 2
- 241000589517 Pseudomonas aeruginosa Species 0.000 description 2
- 239000002253 acid Substances 0.000 description 2
- 238000004458 analytical method Methods 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
- 229940041514 candida albicans extract Drugs 0.000 description 2
- 125000004432 carbon atom Chemical group C* 0.000 description 2
- 238000004587 chromatography analysis Methods 0.000 description 2
- SECPZKHBENQXJG-UHFFFAOYSA-N cis-palmitoleic acid Natural products CCCCCCC=CCCCCCCCC(O)=O SECPZKHBENQXJG-UHFFFAOYSA-N 0.000 description 2
- 238000004440 column chromatography Methods 0.000 description 2
- 238000007796 conventional method Methods 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 239000008344 egg yolk phospholipid Substances 0.000 description 2
- 229940068998 egg yolk phospholipid Drugs 0.000 description 2
- 239000003995 emulsifying agent Substances 0.000 description 2
- 238000001914 filtration Methods 0.000 description 2
- 235000019688 fish Nutrition 0.000 description 2
- 238000005194 fractionation Methods 0.000 description 2
- 238000004817 gas chromatography Methods 0.000 description 2
- QXJSBBXBKPUZAA-UHFFFAOYSA-N isooleic acid Natural products CCCCCCCC=CCCCCCCCCC(O)=O QXJSBBXBKPUZAA-UHFFFAOYSA-N 0.000 description 2
- VLKZOEOYAKHREP-UHFFFAOYSA-N n-Hexane Chemical compound CCCCCC VLKZOEOYAKHREP-UHFFFAOYSA-N 0.000 description 2
- WQEPLUUGTLDZJY-UHFFFAOYSA-N n-Pentadecanoic acid Natural products CCCCCCCCCCCCCCC(O)=O WQEPLUUGTLDZJY-UHFFFAOYSA-N 0.000 description 2
- ZQPPMHVWECSIRJ-KTKRTIGZSA-N oleic acid Chemical compound CCCCCCCC\C=C/CCCCCCCC(O)=O ZQPPMHVWECSIRJ-KTKRTIGZSA-N 0.000 description 2
- 235000019319 peptone Nutrition 0.000 description 2
- 150000003839 salts Chemical class 0.000 description 2
- 235000019512 sardine Nutrition 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- 239000012138 yeast extract Substances 0.000 description 2
- 229920001817 Agar Polymers 0.000 description 1
- 241000894006 Bacteria Species 0.000 description 1
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 description 1
- 241000238366 Cephalopoda Species 0.000 description 1
- 241000195649 Chlorella <Chlorellales> Species 0.000 description 1
- 241000195493 Cryptophyta Species 0.000 description 1
- FBPFZTCFMRRESA-JGWLITMVSA-N D-glucitol Chemical compound OC[C@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-JGWLITMVSA-N 0.000 description 1
- 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 1
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerol Natural products OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 1
- PVNIQBQSYATKKL-UHFFFAOYSA-N Glycerol trihexadecanoate Natural products CCCCCCCCCCCCCCCC(=O)OCC(OC(=O)CCCCCCCCCCCCCCC)COC(=O)CCCCCCCCCCCCCCC PVNIQBQSYATKKL-UHFFFAOYSA-N 0.000 description 1
- 244000068988 Glycine max Species 0.000 description 1
- 235000010469 Glycine max Nutrition 0.000 description 1
- OYHQOLUKZRVURQ-HZJYTTRNSA-N Linoleic acid Chemical compound CCCCC\C=C/C\C=C/CCCCCCCC(O)=O OYHQOLUKZRVURQ-HZJYTTRNSA-N 0.000 description 1
- 229910019142 PO4 Inorganic materials 0.000 description 1
- 241000269851 Sarda sarda Species 0.000 description 1
- 241000269821 Scombridae Species 0.000 description 1
- 241001504592 Trachurus trachurus Species 0.000 description 1
- CWRILEGKIAOYKP-SSDOTTSWSA-M [(2r)-3-acetyloxy-2-hydroxypropyl] 2-aminoethyl phosphate Chemical compound CC(=O)OC[C@@H](O)COP([O-])(=O)OCCN CWRILEGKIAOYKP-SSDOTTSWSA-M 0.000 description 1
- 150000007513 acids Chemical class 0.000 description 1
- 239000008272 agar Substances 0.000 description 1
- 235000021342 arachidonic acid Nutrition 0.000 description 1
- 229940114079 arachidonic acid Drugs 0.000 description 1
- 230000001580 bacterial effect Effects 0.000 description 1
- 239000011575 calcium Substances 0.000 description 1
- 229910052791 calcium Inorganic materials 0.000 description 1
- 159000000007 calcium salts Chemical class 0.000 description 1
- 210000000170 cell membrane Anatomy 0.000 description 1
- UXTMROKLAAOEQO-UHFFFAOYSA-N chloroform;ethanol Chemical compound CCO.ClC(Cl)Cl UXTMROKLAAOEQO-UHFFFAOYSA-N 0.000 description 1
- QSKWJTXWJJOJFP-UHFFFAOYSA-N chloroform;ethoxyethane Chemical compound ClC(Cl)Cl.CCOCC QSKWJTXWJJOJFP-UHFFFAOYSA-N 0.000 description 1
- OQNGCCWBHLEQFN-UHFFFAOYSA-N chloroform;hexane Chemical compound ClC(Cl)Cl.CCCCCC OQNGCCWBHLEQFN-UHFFFAOYSA-N 0.000 description 1
- WORJEOGGNQDSOE-UHFFFAOYSA-N chloroform;methanol Chemical compound OC.ClC(Cl)Cl WORJEOGGNQDSOE-UHFFFAOYSA-N 0.000 description 1
- GVJHHUAWPYXKBD-UHFFFAOYSA-N d-alpha-tocopherol Natural products OC1=C(C)C(C)=C2OC(CCCC(C)CCCC(C)CCCC(C)C)(C)CCC2=C1C GVJHHUAWPYXKBD-UHFFFAOYSA-N 0.000 description 1
- 230000001804 emulsifying effect Effects 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 150000002148 esters Chemical class 0.000 description 1
- STVYPZZQOXPZBX-UHFFFAOYSA-N ethanol;hexane;hydrate Chemical compound O.CCO.CCCCCC STVYPZZQOXPZBX-UHFFFAOYSA-N 0.000 description 1
- 235000021588 free fatty acids Nutrition 0.000 description 1
- 239000000499 gel Substances 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 235000011187 glycerol Nutrition 0.000 description 1
- 230000002209 hydrophobic effect Effects 0.000 description 1
- OYHQOLUKZRVURQ-IXWMQOLASA-N linoleic acid Natural products CCCCC\C=C/C\C=C\CCCCCCCC(O)=O OYHQOLUKZRVURQ-IXWMQOLASA-N 0.000 description 1
- 235000020778 linoleic acid Nutrition 0.000 description 1
- 229960004232 linoleic acid Drugs 0.000 description 1
- 210000004185 liver Anatomy 0.000 description 1
- 235000020640 mackerel Nutrition 0.000 description 1
- 125000002496 methyl group Chemical group [H]C([H])([H])* 0.000 description 1
- 239000010452 phosphate Substances 0.000 description 1
- 229920000642 polymer Polymers 0.000 description 1
- 239000011591 potassium Substances 0.000 description 1
- 229910052700 potassium Inorganic materials 0.000 description 1
- XAEFZNCEHLXOMS-UHFFFAOYSA-M potassium benzoate Chemical compound [K+].[O-]C(=O)C1=CC=CC=C1 XAEFZNCEHLXOMS-UHFFFAOYSA-M 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 235000003441 saturated fatty acids Nutrition 0.000 description 1
- 150000004671 saturated fatty acids Chemical class 0.000 description 1
- 239000013535 sea water Substances 0.000 description 1
- 239000000741 silica gel Substances 0.000 description 1
- 229910002027 silica gel Inorganic materials 0.000 description 1
- 239000000377 silicon dioxide Substances 0.000 description 1
- 159000000000 sodium salts Chemical class 0.000 description 1
- 239000008347 soybean phospholipid Substances 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 125000004079 stearyl group Chemical group [H]C([*])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])[H] 0.000 description 1
- 239000004094 surface-active agent Substances 0.000 description 1
- 229960001295 tocopherol Drugs 0.000 description 1
- 229930003799 tocopherol Natural products 0.000 description 1
- 235000010384 tocopherol Nutrition 0.000 description 1
- 239000011732 tocopherol Substances 0.000 description 1
- UFTFJSFQGQCHQW-UHFFFAOYSA-N triformin Chemical compound O=COCC(OC=O)COC=O UFTFJSFQGQCHQW-UHFFFAOYSA-N 0.000 description 1
- PHYFQTYBJUILEZ-IUPFWZBJSA-N triolein Chemical compound CCCCCCCC\C=C/CCCCCCCC(=O)OCC(OC(=O)CCCCCCC\C=C/CCCCCCCC)COC(=O)CCCCCCC\C=C/CCCCCCCC PHYFQTYBJUILEZ-IUPFWZBJSA-N 0.000 description 1
- 238000009736 wetting Methods 0.000 description 1
- GVJHHUAWPYXKBD-IEOSBIPESA-N α-tocopherol Chemical compound OC1=C(C)C(C)=C2O[C@@](CCC[C@H](C)CCC[C@H](C)CCCC(C)C)(C)CCC2=C1C GVJHHUAWPYXKBD-IEOSBIPESA-N 0.000 description 1
Landscapes
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
Description
【発明の詳細な説明】 〔産業上の利用分野〕 本発明は、微生物から高度不飽和脂肪酸リン脂 質を製造する方法に関するものである。該リン脂 質は食品、化粧品、医薬品、農業、水産、化成品 などの分野において有用である。TECHNICAL FIELD The present invention relates to a method for producing a highly unsaturated fatty acid phosphate from a microorganism. The phospholipids are useful in fields such as foods, cosmetics, pharmaceuticals, agriculture, fisheries, and chemical products.
リン脂質は、高等動植物をはじめ各種生物中に 含まれ、主として細胞膜の構成物として生体中で 重要な働きをしている。このうち現在、工業的規 模で生産されているリン脂質は、大豆リン脂質と 卵黄リン脂質である。これらは天然界面活性剤で あり、この乳化作用、分散作用、湿潤作用などを 利用して食品、化粧品、医薬品などの分野で使用 されている。しかしながら、かかる大豆および卵 黄リン脂質をはじめとする高等動植物を起源とす るリン脂質は、その構成脂肪酸が炭素数１６〜 １８の飽和脂肪酸及び／又はモノエン脂肪酸を主 成分とするものであり、炭素数１８かつ不飽和結 合２以上の不飽和脂肪酸（以下、高度不飽和脂肪 酸と称す。）リン脂質が工業的に単離された例は 見当たらない。 Phospholipids are contained in various organisms such as higher animals and plants, and mainly play an important role in the living body as a constituent of cell membranes. Of these, the phospholipids currently produced on an industrial scale are soybean phospholipids and egg yolk phospholipids. These are natural surfactants, which are used in the fields of food, cosmetics, pharmaceuticals, etc. by utilizing their emulsifying action, dispersing action and wetting action. However, phospholipids originating from higher animals and plants such as soybeans and egg yolk phospholipids are those whose constituent fatty acids contain saturated fatty acids having 16 to 18 carbon atoms and / or monoene fatty acids as main components, An unsaturated fatty acid having 18 carbon atoms and an unsaturated bond of 2 or more (hereinafter referred to as a polyunsaturated fatty acid) phospholipid has not been industrially isolated.
本発明の目的は、高度不飽和脂肪酸リン脂質を 工業的に効率良く、短時間で、容易に得ることの できる製造法を提供することにある。 An object of the present invention is to provide a method for producing a highly unsaturated fatty acid phospholipid industrially efficiently, in a short time, and easily.
〔課題を解決するための手段〕 本発明者らは、上記の目的に合った製造法を見 出すために鋭意研究の結果、培地中に高度不飽和 脂肪酸及び／又はその誘導体を含む油脂を添加し て培養した微生物の脂質について、次のような知 見を得た。即ち、微生物の総脂質含有量は乾物 換算で６〜１２％と高く、そのうち９０％以上が リン脂質であること、リン脂質の脂肪酸組成を 見ても高度不飽和脂肪酸が１０〜４０％と高いこ と、本発明で用いる微生物の増殖速度は、クロ レラ等藻類に比べて著しく速く、多量の菌体を極 めて短時間のうちに得ることができる。[Means for Solving the Problems] The inventors of the present invention have conducted extensive studies as a result of finding out a production method suitable for the above purpose, and as a result, added an oil or fat containing a highly unsaturated fatty acid and / or a derivative thereof to a medium. The following findings were obtained regarding the lipids of the cultivated microorganisms. That is, the total lipid content of microorganisms is as high as 6 to 12% in terms of dry matter, 90% or more of which is phospholipids, and the fatty acid composition of phospholipids is as high as 10 to 40%. In addition, the growth rate of the microorganism used in the present invention is remarkably faster than that of algae such as Chlorella, and a large amount of cells can be obtained within a short time.
本発明は、以上の知見に基づいて完成されたも ので、微生物を高度不飽和脂肪酸及び／又はその 誘導体を含む油脂を添加した培地中にて培養し、 次いでこれら培養した微生物から抽出分離するこ とを特徴とする高度不飽和脂肪酸リン脂質の製造 法である。 The present invention has been completed based on the above findings. Therefore, the microorganisms are cultured in a medium supplemented with oils and fats containing highly unsaturated fatty acids and / or their derivatives, and then extracted and separated from these cultured microorganisms. And a method for producing a highly unsaturated fatty acid phospholipid characterized by
本発明で用いる高度不飽和脂肪酸及び／又はそ の誘導体を含む油脂としては、好ましくは総脂肪 酸中の高度不飽和脂肪酸の占める割合が１０％以 上のものが良く、このようなものの例をあげると イワシ、サバ、アジ等の青背魚より抽出した油脂、 マグロやカツオ等の大型海産魚の眼窩脂肪由来の 油脂、微生物由来の油脂、イカの肝臓より抽出し た海産物由来の油脂などが好ましい例としてあげ られる。 The fats and oils containing a polyunsaturated fatty acid and / or its derivative used in the present invention preferably have a proportion of polyunsaturated fatty acids in total fatty acids of 10% or more. Preferred are oils and fats extracted from blue-backed fish such as sardines, mackerel, horse mackerel, oils and fats derived from orbital fat of large marine fish such as tuna and bonito, oils and fats derived from microorganisms, and oils and fats derived from marine products extracted from squid liver. Take as an example.
本発明で使用できる微生物は、特に属、種ある いは株などを限定するものではないが、通常は、 エシェリシア（Escherichia）属、セラチア （Serratia）属、バチルス（Bacillus）属、スタ フェロコッカス（Staphylococcus）属、シュード モナス（Pseudomonas）属、アルテロモナス （Alteromonas）属、シーワネラ（Shewanella）属 などに分類される微生物等を用いることができる。 Microorganisms that can be used in the present invention are not particularly limited to genus, species, strains, etc., but are usually Escherichia genus, Serratia genus, Bacillus genus, Staferococcus ( Staphylococcus genus, Pseudomonas genus, Alteromonas genus, Shewanella genus, etc. can be used.
これらの微生物については公的微生物寄託機関等 にて容易に入手できる。又新種微生物微工研菌寄 第１１６７１号を用いることが出来る。These microorganisms can be easily obtained at public microorganism depository institutions. In addition, New Microorganisms Microbiology Research Institute No. 11671 can be used.
上記のエシェリアに属する微生物の例として、 エシェリア・コリ（Escherichia coli）ＩＦＯ −３３０１セラチアに属する微生物の例としてセ ラチア・マルセッセンス（Serratia marcescens） ＩＦＯ−３０５４、バチルスに属する微生物の例 としてバチルス・セレウス（Bacillus sereus） ＩＦＯ−３００１及びバチルス・サブチリス （Bacillus subtilis）ＩＡＭ−１０２６、スタ フィロコッカスに属する微生物の例としてスタフ ィロコッカス・オーレウス（Staphylococcus aureus ）ＡＴＣＣ−１２６００、シュードモナス に属する微生物の例としてシュードモナス・アエ ルギノーサ（Pseudomonas aeruginosa）ＩＦＯ−３９
１８、シュードモナス・ピュートリファシ エンス（Peudomonas，putrefaciens）SCRC−2181， （FERMBP−2917），SCRC−2201（FERMBP−2916）， SCRC−2271（FERMBP−2915），SCRC− 2341 （FERMBP−2918），SCRC−2451（FERMBP−2919）， SCRC−2642（FERMBP−2920），SCRC− 2792（FERMB −2921），SCRC−2878（FERMBP−1623），SCRC− 3011（FERMBP−2913），SCRC−3022（ FERMBP− 2914）を挙げることができる。Examples of the above-mentioned microorganisms belonging to Escherichia include Escherichia coli IFO-3301, examples of microorganisms belonging to Serratia marcescens IFO-3054, and examples of microorganisms belonging to Bacillus cereus (E. coli ). Bacillus sereus ) IFO-3001 and Bacillus subtilis IAM-1026, Staphylococcus aureus ATCC-12600 as an example of a microorganism belonging to Staphylococcus , and Pseudomonas aerus as an example of a microorganism belonging to Pseudomonas. ( Pseudomonas aeruginosa ) IFO-39
18, Pseudomonas Pyutorifashi Enns (Peudomonas, putrefaciens) SCRC-2181 , (FERMBP-2917), SCRC-2201 (FERMBP-2916), SCRC-2271 (FERMBP-2915), SCRC- 2341 (FERMBP-2918), SCRC-2451 (FERMBP-2919), SCRC-2642 (FERMBP-2920), SCRC-2792 (FERMB-2921), SCRC-2878 (FERMBP-1623), SCRC-3011 (FERMBP-2913), SCRC-3022 (FERMBP) -2914).
アルテロモナスに属する微生物の例として、ア ルテロモナス・ピュートリファシエンス （Alteromonas putrefaciens）SCRC−2871 （FERMBP−1624）及びアルテロモナス・ピュート リファシエンス・サブスピーシズ・サガミファシ エンス（Alteromonas putrefaciens subspecies sagamifaciens）SCRC−1162（FERMBP −1626）等を挙げることができる。Examples of microorganisms belonging to Alteromonas are Alteromonas putrefaciens SCRC-2871 (FERMBP-1624) and Alteromonas putrefaciens subspecies sagamifaciens subspecies sagamifaciens SCRC-1162 (FERMBP-1624). ) Etc. can be mentioned.
シーワネラに属する微生物の例として、シーワ ネラ・ピュートリファシエンス（Shewanella putrefaciens ）SCRC−2874（FERMBP−1625）等を 挙げることができる。Examples of microorganisms belonging to Shiwanella include Shewanella putrefaciens SCRC-2874 (FERMBP-1625) and the like.
本発明の実施に当たっては、微生物を常法によ り例えば培地で培養し、これを遠心分離法などで 集め、洗浄、乾燥して微生物粉末を得る。この際 の培地としては例えば次の第１表に示す組成の培 地を調整することができる。 In carrying out the present invention, microorganisms are cultured by a conventional method, for example, in a medium, collected by a centrifugation method, washed and dried to obtain a microorganism powder. As the medium at this time, for example, a medium having the composition shown in Table 1 below can be prepared.
又微生物に海洋性細菌を用いる場合には、次の 第２表に示す組成の培地を調整することができる。 When marine bacteria are used as the microorganism, a medium having the composition shown in Table 2 below can be prepared.
培地中に添加する油脂としては、脂肪酸の遊離 型、エステル型、塩等の誘導体を含む油脂を用い る事ができる。より好ましくは高度不飽和脂肪酸 の塩、例えばカリウム塩、ナトリウム塩、カルシ ウム塩等を含む油脂を用いる事ができる。 As the oil / fat added to the medium, an oil / fat containing a free fatty acid type, an ester type, or a derivative such as a salt can be used. More preferably, fats and oils containing salts of highly unsaturated fatty acids, such as potassium salt, sodium salt, calcium salt and the like can be used.
添加する油脂は、培地に対し０．００１％〜 １０％、好ましくは０．０１％〜１．０％を添加 することが、微生物の増殖及び高度不飽和脂肪酸 の取り込み率から見て望ましい。また微生物の培 養時に乳化剤等を添加しても良い。 It is desirable to add 0.001% to 10%, preferably 0.01% to 1.0%, of the fats and oils to the medium from the viewpoint of the growth of microorganisms and the uptake rate of highly unsaturated fatty acids. In addition, an emulsifier or the like may be added during cultivation of the microorganism.
この微生物粉末から、有機溶剤などを用いて脂 質抽出を行い、脱溶剤後、アセトン分画を行い、 アセトンに不溶な物質を集めれば、リン脂質の粗 分画物が得ることができる。有機溶剤としては、 リン脂質を溶解する通常の溶剤を単独又は混合し て用いられることができ、例えば、クロロホルム −メタノール系、ｎ−ヘキサン−エタノール−水 系、クロロホルム−ヘキサン系、クロロホルム− エーテル系、クロロホルム−エタノール系などが 挙げられる。 A crude phospholipid fraction can be obtained by subjecting the microbial powder to fat extraction using an organic solvent or the like, desolventization, acetone fractionation, and collection of acetone-insoluble substances. As the organic solvent, an ordinary solvent that dissolves phospholipids can be used alone or in combination, and examples thereof include chloroform-methanol system, n-hexane-ethanol-water system, chloroform-hexane system, chloroform-ether system. , Chloroform-ethanol system and the like.
また、分別に用いられる溶剤は、通常、脂質の 分別に用いられる有機溶剤を用いることができる。 As the solvent used for separation, an organic solvent usually used for separation of lipids can be used.
例えば、アセトン、酢酸、クロロホルム、エーテ ルなどが挙げられる。また、溶剤にMgCl2，カル シウムなどを添加し、その共存下、分別を行って もよく、必要に応じて加熱、冷却等を行っても良 い、さらに、得られたすべての画分をカラムクロ マトグラフィーで再分画し、リン脂質の画分を集 めると高純度のリン脂質が高収率で得られる。更 に、通常の方法で精製することにより、単離する ことが出来る。カラムクロマトグラフィーとして は、例えば、疏水性クロマトグラフィー、シリカ 系クロマトグラフィー、トヨパール（商品名 東 ソー（株）製）等のポリマーゲル、シリカゲルに オクタデシル基を結合させたものなどを用いるこ とができる。Examples include acetone, acetic acid, chloroform, ether and the like. It is also possible to add MgCl 2 , calcium, etc. to the solvent and perform fractionation in the coexistence thereof, and to heat and cool as necessary, and further to obtain all the obtained fractions. High-purity phospholipids can be obtained in high yield by re-fractionating by column chromatography and collecting the phospholipid fractions. Further, it can be isolated by purification by a usual method. Examples of column chromatography that can be used include hydrophobic chromatography, silica-based chromatography, polymer gels such as Toyopearl (trade name, manufactured by Tosoh Corporation), and silica gel having an octadecyl group bonded thereto. ..
このようにして得られるリン脂質は、ＴＬＣ （薄層クロマトグラフィー）でその組成を分析し、 さらに常法によりケン化分解、メチルエステル化 してＧＬＣ（ガスクロマトグラフィー）で分析し て構成脂肪酸組成を求めることができる。 The composition of the thus obtained phospholipids is analyzed by TLC (thin layer chromatography), further saponified by a conventional method, methyl esterified and analyzed by GLC (gas chromatography) to determine the constituent fatty acid composition. Can be asked.
上述の方法により得られるリン脂質の組成物は、 ホスファチジルエタノールアミン、リゾホスファ チジルエタノールアミン、ホスファチジルグリセ ロール、カルジオリピンなどであり、またその構 成脂肪酸はドコサヘキサエン酸、エイコサペンチ エン酸、アラキドン酸、リノール酸、オレイン酸、 パルミトレイン、パルチミン酸、ミリスチン酸な どである。 The composition of the phospholipid obtained by the above method is phosphatidylethanolamine, lysophosphatidylethanolamine, phosphatidylglycerol, cardiolipin, etc., and its constituent fatty acids are docosahexaenoic acid, eicosapentaenoic acid, arachidonic acid, linoleic acid. , Oleic acid, palmitolein, palmitic acid, myristic acid.
実施例 １ エシェリシア・コリ（Escherichia coli） ＩＦＯ−３３０１を酵母エキス０．５％、ペプト ン１％、マグロ眼窩脂肪より抽出して得られた脂 肪酸カリウム塩からなる油脂（ドコサヘキサエン 酸３０％、エイコサペンタエン酸８．４％、含有） ０．１％、グルコース０．５％を含有し、pH７に 調整した培地中で１０ｌ好気培養し、細胞を遠心 分離して集め、洗浄、乾燥して微生物粉末を得た。Example 1 Escherichia coli IFO-3301 0.5% yeast extract, 1% peptone, oil and fat composed of potassium fatty acid salt obtained by extracting from tuna orbital fat (docosahexaenoic acid 30%, Eicosapentaenoic acid 8.4%, containing) 0.1% agar, 0.5% glucose 0.5%, aerobically cultivated 10 l in a medium adjusted to pH 7, collect cells by centrifugation, wash and dry A microbial powder was obtained.
微生物の乾燥粉末１００ｇをクロロホルム：メタ ノール＝２：１混合溶剤で抽出し、８ｇの抽出脂 質を得た。この脱溶剤後の総脂質にアセトン400 ｍｌを加え、冷却しながら撹拌を行い、糖脂質、 キノンを分別し濾過によりアセトン不溶分を回収 した。得られたアセトン不溶分をクロロホルムに 溶解し、次にこの溶液をシリカゲルカラムクロマ トグラフィーにかけ、クロロホルム、クロロホル ム：メタノール＝２：１、クロロホルム：メタノ ール＝１：１およびメタノールで順次展開した。100 g of dried microorganism powder was extracted with a mixed solvent of chloroform: methanol = 2: 1 to obtain 8 g of extracted fat. Acetone 400 ml was added to the total lipids after the solvent removal, and the mixture was stirred while cooling, the glycolipids and quinone were separated, and the acetone insoluble content was collected by filtration. The obtained acetone-insoluble matter was dissolved in chloroform, and this solution was then subjected to silica gel column chromatography, and developed sequentially with chloroform, chloroform: methanol = 2: 1, chloroform: methanol = 1: 1 and methanol. ..
得られた各フラクションをＴＬＣでチェックし、 目的とするリン脂質のフラクションをいくつか見 出し、それらを回収し、脱溶剤したところ約７ｇ のリン脂質が得られた。得られたリン脂質の組成 はＴＬＣ分析の結果、ホスファチジルグリセロー ル（７０％）およびホスファチジルエタノールア ミン（２０％）が主成分であり、またこれらの脂 質の構成高度不飽和脂肪酸はドコサヘキサエン酸 １０．５％、エイコサペンタエン酸4.８％であっ た。Each of the obtained fractions was checked by TLC to find some fractions of the desired phospholipid, which were collected and desolvated to obtain about 7 g of phospholipid. As a result of TLC analysis, the composition of the obtained phospholipids was phosphatidylglycerol (70%) and phosphatidylethanolamine (20%) as main components, and the highly unsaturated fatty acids constituting these fats were docosahexaenoic acid. It was 10.5% and eicosapentaenoic acid 4.8%.
実施例 ２ セラチア・マルセッセンス（Serratia marcescens ）ＩＦＯ−３０５４を上記実施例１と 同様の方法で培養して得た該微生物菌体を実施例 １と同方法にて抽出したところ、リン脂質を得た。Was extracted in Example 2 Serratia marcescens (Serratia marcescens) IFO-3054 of the above Example 1 and the same method in culture obtained was microorganism cells Example 1 and the same methods, to obtain a phospholipid ..
該リン脂質の構成高度不飽和脂肪酸は、ドコサヘ キサエン酸２．５％、エイコサペンタエン酸０％ であった。The constituent polyunsaturated fatty acids of the phospholipid were docosahexaenoic acid 2.5% and eicosapentaenoic acid 0%.
実施例 ３ バチルス・セレウス（Bacillus cereus） ＩＦＯ−３００１を上記実施例１と同様の方法で 培養して得た該微生物菌体を実施例１と同方法に て抽出したところ、リン脂質を得た。該リン脂質 の構成高度不飽和脂肪酸は、ドコサヘキサエン酸 ８．９％、エイコサペンタエン酸１２．０％であ った。Example 3 Bacillus cereus IFO-3001 was cultured in the same manner as in Example 1 above, and the obtained microbial cells were extracted in the same manner as in Example 1 to obtain phospholipids. .. The constituent polyunsaturated fatty acids of the phospholipid were 8.9% docosahexaenoic acid and 12.0% eicosapentaenoic acid.
実施例 ４ バチルス・サブチリス（Bacillus subtilis ＩＡＭ−１０２６を上記実施例１と同様の方法で 培養して得た該微生物菌体を実施例１と同方法に て抽出したところ、リン脂質を得た。該リン脂質 の構成高度不飽和脂肪酸は、ドコサヘキサエン酸 ２２．４％、エイコサペンタエン酸１４．９％で あった。Example 4 Bacillus subtilis IAM-1026 was cultured in the same manner as in Example 1 above, and the obtained microbial cells were extracted in the same manner as in Example 1 to obtain a phospholipid. The constituent polyunsaturated fatty acids of the phospholipid were docosahexaenoic acid (22.4%) and eicosapentaenoic acid (14.9%).
実施例 ５ スタフィロコッカス・オーレウス （Staphylococcus aureus）ＡＴＣＣ−12600 を上記実施例１と同様の方法で培養して得た該微 生物菌体を実施例１と同方向にて抽出したところ、 リン脂質を得た。該リン脂質の構成高度不飽和脂 肪酸は、ドコサヘキサエン酸３．８％、エイコサ ペンタエン酸２．４％であった。Example 5 Staphylococcus aureus ATCC-12600 was cultured in the same manner as in Example 1 above, and the obtained microorganism cells were extracted in the same direction as in Example 1. Got The constituent highly unsaturated fatty acids of the phospholipids were docosahexaenoic acid 3.8% and eicosapentaenoic acid 2.4%.
実施例 ６ シュードモナス・アエルギノーサ（Pseudomonas aeruginosa ）ＩＦＯ−３９１８を上記実施例１と 同様の方法で培養して得た該微生物菌体を実施例 １と同方法にて抽出したところ、リン脂質を得た。Example 6 The microbial cells obtained by culturing Pseudomonas aeruginosa IFO-3918 in the same manner as in Example 1 above were extracted by the same method as in Example 1 to obtain phospholipids. ..
該リン脂質の構成高度不飽和脂肪酸は、ドコサヘ キサエン酸６．９％、エイコサペンタエン酸1.８ ％であった。The constituent polyunsaturated fatty acids of the phospholipid were 6.9% docosahexaenoic acid and 1.8% eicosapentaenoic acid.
実施例 ７ 新種海洋細菌微工研菌寄第１１６７１号を上記 実施例１と同様の方法で培養して得た該微生物菌 体を実施例１と同方法にて抽出したところ、リン 脂質を得た。該リン脂質の構成高度不飽和脂肪酸 は、ドコサヘキサエン酸２０．４％、エイコサペ ンタエン酸１８．４％であった。Example 7 A microbial cell obtained by culturing New Marine Bacterial Microbiology Research Institute No. 11671 in the same manner as in Example 1 above was extracted in the same manner as in Example 1 to obtain a phospholipid. It was The constituent highly unsaturated fatty acids of the phospholipid were docosahexaenoic acid 20.4% and eicosapentaenoic acid 18.4%.
実施例 ８ 海洋微生物（Pseudomonas putrefaciens SCRC−2878(FERMBP−1623)）を酵母エキス０．５ ％、ペプトン１％、イワシより抽出した油成分 ０．１％（トリグリセライドを主成分とし、 ＤＨＡを１０％含有）、乳化剤（トコフェロール １５％、Ｄ−ソルビット４５％、グリセリン脂肪 酸エステル３％を含有）０．００３％を含有し、 ｐＨ７に調整した海水培地中で１０ｌ好気培養し、 細胞を遠心分離して集め、洗浄、乾燥して海洋微 生物粉末を得た。海洋微生物の乾燥粉末１００ｇ をクロロホルム：メタノール＝２：１混合溶剤で 抽出し、８ｇの抽出脂質を得た。この脱溶剤後の 総脂質にアセトン４００ｍｌを加え、冷却しなが ら撹拌を行い、糖脂質、キノンを分別をし濾過に よりアセトン不溶分を回収した。得られたアセト ン不溶分をクロロホルムに溶解し、次にこの溶液 をシリカゲルカラムクロマトグラフィーにかけ、 クロロホルム、クロロホルム：メタノール＝２： １、クロロホルム：メタノール＝１：１およびメ タノールで順次展開した。得られた各フラクショ ンをＴＬＣでチェックし、目的とするリン脂質の フラクションをいくつか見出し、それらを回収し、 脱溶剤したところ約７ｇのリン脂質組成物が得ら れた。得られたリン脂質の組成はＴＬＣ分析の結 果、ホスファチジルグリセロールおよびホスファ チジルエタノールアミンが主成分であり、また構 成脂肪酸はＤＨＡ（１７．５％）、ＥＰＡ （６．８％）、オレイン酸（６．２％）、パルミ チン酸（８．６％）、パルミトレイン酸（８．０ ％）などであった。Example 8 A marine microorganism ( Pseudomonas putrefaciens SCRC-2878 (FERM BP-1623)) yeast extract 0.5%, peptone 1%, oil component 0.1% extracted from sardines (triglyceride as a main component, DHA 10%) Containing), an emulsifier (containing 15% tocopherol, 45% D-sorbit, 3% glycerin fatty acid ester) 0.003%, aerobically culturing 10 l in a seawater medium adjusted to pH 7, and centrifuging the cells. , Collected, washed and dried to obtain marine organism powder. 100 g of dried powder of marine microorganism was extracted with a mixed solvent of chloroform: methanol = 2: 1 to obtain 8 g of extracted lipid. 400 ml of acetone was added to the total lipids after the solvent removal, and the mixture was stirred while cooling, the glycolipids and quinone were separated, and the acetone insoluble content was collected by filtration. The aceton-insoluble matter thus obtained was dissolved in chloroform, and this solution was then subjected to silica gel column chromatography, followed by sequential development with chloroform, chloroform: methanol = 2: 1, chloroform: methanol = 1: 1, and methanol. Each of the obtained fractions was checked by TLC to find some fractions of the target phospholipid, which were collected and desolvated to obtain about 7 g of the phospholipid composition. As a result of TLC analysis, the composition of the obtained phospholipids consisted mainly of phosphatidylglycerol and phosphatidylethanolamine, and the constituent fatty acids were DHA (17.5%), EPA (6.8%) and olein. Acid (6.2%), palmitic acid (8.6%), palmitoleic acid (8.0%) and the like.
実施例 ９ 海洋微生物（Alteromonas putrefaciens SCRC−2871（FERMBP−1624)）を上記実施例８と同 様の培地中で培養して細胞を得、遠心集菌、洗浄、 乾燥して海洋微生物粉末を得た。該海洋微生物粉 末１１０ｇをｎ−ヘキサン：エタノール：水＝２ ：０．９：０．１混合溶剤で抽出し、１２ｇの総 脂質を得た。該総脂質にアセトン５００ｍｌを加 え、冷却下に撹拌して、糖脂質、キノンを分別し、 濾過してアセトン不溶分を回収した。得られたア セトン不溶分をブタノールに溶解し、展開溶媒と してブタノール、酢酸および水を用い、実施例１ と同様にシリカゲルカラムクロマトグラフィーで 分画した。各フラクションをＴＬＣでチェックし ながら、リン脂質画分を集め、リン脂質組成物 １０．５ｇを得た。該リン脂質組成物はホスファ チジルグリセロールおよびホスファチジルエタノ ールアミンを主成分とし、また構成脂肪酸は ＤＨＡ（２５．４％）、ＥＰＡ（７．６％）、オ レイン酸（２．４％）、パルミチン酸（６．８％）、 パルミトレイン酸（１１．６％）、ミリスチン酸 （２．５％）、などであった。Example 9 Marine microorganisms ( Alteromonas putrefaciens SCRC-2871 (FERM BP-1624)) were cultured in the same medium as in Example 8 to obtain cells, which were collected by centrifugation, washed and dried to obtain marine microorganism powder. It was 110 g of the marine microbial powder powder was extracted with a mixed solvent of n-hexane: ethanol: water = 2: 0.9: 0.1 to obtain 12 g of total lipid. Acetone (500 ml) was added to the total lipid, and the mixture was stirred under cooling to separate glycolipids and quinone, and filtered to collect acetone-insoluble matter. The obtained aceton-insoluble matter was dissolved in butanol and subjected to silica gel column chromatography in the same manner as in Example 1 using butanol, acetic acid and water as developing solvents. While checking each fraction by TLC, the phospholipid fraction was collected to obtain 10.5 g of a phospholipid composition. The phospholipid composition is mainly composed of phosphatidylglycerol and phosphatidylethanolamine, and the constituent fatty acids are DHA (25.4%), EPA (7.6%), oleic acid (2.4%) and palmitin. Acids (6.8%), palmitoleic acid (11.6%), myristic acid (2.5%), etc.
本発明の効果は次のようである。 The effects of the present invention are as follows.
高度不飽和脂肪酸及び／又はその誘導体を含む 油脂を添加した培地中で培養した微生物の脂質か ら高度不飽和脂肪酸リン脂質を効率良く、短時間 で且つ容易に単離、精製することが可能となった。 Highly unsaturated fatty acid phospholipids can be efficiently and easily isolated and purified from lipids of microorganisms cultured in a medium containing fats and oils containing highly unsaturated fatty acids and / or their derivatives. became.
フロントページの続き (51)Int.Cl.5 識別記号 庁内整理番号 ＦＩ 技術表示箇所 (Ｃ１２Ｐ 9/00 Ｃ１２Ｒ 1:085） (Ｃ１２Ｐ 9/00 Ｃ１２Ｒ 1:43） (Ｃ１２Ｐ 9/00 Ｃ１２Ｒ 1:445） (Ｃ１２Ｐ 9/00 Ｃ１２Ｒ 1:125） (Ｃ１２Ｐ 9/00 Ｃ１２Ｒ 1:385） (Ｃ１２Ｐ 9/00 Ｃ１２Ｒ 1:01） (Ｃ１２Ｐ 9/00 Ｃ１２Ｒ 1:38） Continuation of the front page (51) Int.Cl. 5 Identification number Office reference number FI technical display location (C12P 9/00 C12R 1: 085) (C12P 9/00 C12R 1:43) (C12P 9/00 C12R 1: 445) (C12P 9/00 C12R 1: 125) (C12P 9/00 C12R 1: 385) (C12P 9/00 C12R 1:01) (C12P 9/00 C12R 1:38)
Claims (12)
の誘 導体を含む油脂を添加した培地中にて培養し、 次いでこれら培養した微生物から抽出分離する ことを特徴とする高度不飽和脂肪酸リン脂質の 製造法。1. A polyunsaturated fatty acid phospholipid characterized by culturing a microorganism in a medium containing an oil or fat containing a highly unsaturated fatty acid and / or its derivative, and then extracting and separating from the cultivated microorganism. Manufacturing method.
属、セ ラチア（Serratia）属、バチルス（Bacillus） 属、スタフィロコッカス（Staphylococcus）属、 シュードモナス（Pseudomonas）属、アルテロモ ナス（Alteromonas）属又はシーワネラ （Shewanella）属である特許請求の範囲第(1)項 に記載の製造法。2. The microorganism is Escherichia.
A genus, a Serratia genus, a Bacillus genus, a Staphylococcus genus, a Pseudomonas genus, an Alteromonas genus, or a Shewanella genus. Claims (1) The manufacturing method as described in the section above.
る特 許請求の範囲第(1)項の記載の製造法。3. The method according to claim 1, wherein the microorganism is Microbiology Research Institute No. 11671.
ア・ コリ（Eschelichia Coli）ＩＦＯ−３３０１ である特許請求の範囲第(2)項に記載の製造法。4. The method according to claim 2, wherein the microorganism belonging to the genus Escherichia is Eschelichia Coli IFO-3301.
ルセ ッセンス（Serratia marcescens）IFO-3054 である特許請求の範囲第(2)項に記載の製造法。5. The production method according to claim (2), wherein the microorganism of the genus Serratia is Serratia marcescens IFO-3054.
レウ ス（Bacillus cereus）ＩＦＯ−３００１又は バチルス・サブチリス（Bacillus subtilis） ＩＡＭ−１０２６である特許請求の範囲第(2)項 に記載の製造法。6. The method according to claim 2, wherein the Bacillus microorganism is Bacillus cereus IFO-3001 or Bacillus subtilis IAM-1026.
タフ ィロコッカス・オーレウス（Staphylococcus aureus ）ＡＴＣＣ−１２６００である特許請求 の範囲第(2)項に記載の製造法。7. The method according to claim 2, wherein the microorganism belonging to the genus Staphylococcus is Staphylococcus aureus ATCC-12600.
ドモ ナス・アエルギノーサ（Pseudomonas aerceginosa） ＩＦＯ−３９１８である特許請求 の範囲第(2)項に記載の製造法。8. The method according to claim (2), wherein the Pseudomonas microorganism is Pseudomonas aerceginosa IFO-3918.
モナ ス・ピュートリファシエンス（Pseudomonas putrefaciens）SCRC−2181，SCRC−2201，SCRC −2271，SCRC−2341，SCRC−2451，SCRC−2642， SCRC−2792，SCRC−2878，SCRC−3011又はSCRC −3022である特許請求の範囲第(2)項に記載の製 造法。9. The Pseudomonas putrefaciens SCRC-2181, SCRC-2201, SCRC-2271, SCRC-2341, SCRC-2451, SCRC-2642, SCRC-2792, SCRC. -2878, SCRC-3011 or SCRC-3022, The manufacturing method according to claim (2).
ロモナ ス・ピュートリファシエンス（Alteromonas putrefaciens ）SCRC−2871又はアルテロモナス ・ピュートリファシエンス・サブスピーシズ・ サガミファシエンス（Alteromonas putrefaciens subspecies sagamifaciens） SCRC−1162である特許請求の範囲第(2)項に記載 の製造法。10. A patent in which the microorganism belonging to the genus Alteromonas is Alteromonas putrefaciens SCRC-2871 or Alteromonas putrefaciens subspecies saga mifaciens SCRC-1162. The manufacturing method according to claim (2).
・ピュ ートリファシエンス（Shewanella putrefaciens ）SCRC−2874である特許請求の範 囲第(2)に記載の製造法。11. The production method according to claim (2), wherein the microorganism belonging to the genus Shiwanella is Shewanella putrefaciens SCRC-2874.
チジル グリセロール、ホスファチジルエタノールアミ ン及び／又はカルジオリピンを主成分とするも のである特許請求の範囲第(1)項記載の製造法。12. The method according to claim 1, wherein the polyunsaturated fatty acid phospholipid contains phosphatidyl glycerol, phosphatidyl ethanolamine and / or cardiolipin as a main component.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP32967390A JPH05123176A (en) | 1990-07-06 | 1990-11-30 | Production of highly unsaturated fatty acid phospholipid |
Applications Claiming Priority (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP17729790 | 1990-07-06 | ||
| JP2-177297 | 1990-07-06 | ||
| JP32967390A JPH05123176A (en) | 1990-07-06 | 1990-11-30 | Production of highly unsaturated fatty acid phospholipid |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| JPH05123176A true JPH05123176A (en) | 1993-05-21 |
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ID=26497894
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP32967390A Pending JPH05123176A (en) | 1990-07-06 | 1990-11-30 | Production of highly unsaturated fatty acid phospholipid |
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| Country | Link |
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| JP (1) | JPH05123176A (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1998001536A1 (en) * | 1996-07-03 | 1998-01-15 | Sagami Chemical Research Center | Microorganisms producing docosahexaenoic acid and process for the production of docosahexaenoic acid |
| US6346276B1 (en) | 1997-10-24 | 2002-02-12 | Asahi Kasei Kabushiki Kaisha | Composition containing useful substances originating in fishes and shellfishes and process for the preparation of the substances |
| CN111057667A (en) * | 2019-12-24 | 2020-04-24 | 天津科技大学 | Complex microbial inoculant and preparation method and application thereof |
-
1990
- 1990-11-30 JP JP32967390A patent/JPH05123176A/en active Pending
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1998001536A1 (en) * | 1996-07-03 | 1998-01-15 | Sagami Chemical Research Center | Microorganisms producing docosahexaenoic acid and process for the production of docosahexaenoic acid |
| US6346276B1 (en) | 1997-10-24 | 2002-02-12 | Asahi Kasei Kabushiki Kaisha | Composition containing useful substances originating in fishes and shellfishes and process for the preparation of the substances |
| CN111057667A (en) * | 2019-12-24 | 2020-04-24 | 天津科技大学 | Complex microbial inoculant and preparation method and application thereof |
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