JPH07289272A - Production of oils and fats highly containing docosahexaenoic acid - Google Patents
Production of oils and fats highly containing docosahexaenoic acidInfo
- Publication number
- JPH07289272A JPH07289272A JP6081654A JP8165494A JPH07289272A JP H07289272 A JPH07289272 A JP H07289272A JP 6081654 A JP6081654 A JP 6081654A JP 8165494 A JP8165494 A JP 8165494A JP H07289272 A JPH07289272 A JP H07289272A
- Authority
- JP
- Japan
- Prior art keywords
- oils
- oil
- docosahexaenoic acid
- fat
- fats
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
- 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 title claims abstract description 85
- 235000020669 docosahexaenoic acid Nutrition 0.000 title claims abstract description 69
- 229940090949 docosahexaenoic acid Drugs 0.000 title claims abstract description 16
- 238000004519 manufacturing process Methods 0.000 title claims abstract description 10
- 235000014593 oils and fats Nutrition 0.000 title abstract description 8
- VTYYLEPIZMXCLO-UHFFFAOYSA-L Calcium carbonate Chemical compound [Ca+2].[O-]C([O-])=O VTYYLEPIZMXCLO-UHFFFAOYSA-L 0.000 claims abstract description 20
- 241000235036 Debaryomyces hansenii Species 0.000 claims abstract description 20
- 238000012258 culturing Methods 0.000 claims abstract description 12
- BWHMMNNQKKPAPP-UHFFFAOYSA-L potassium carbonate Chemical compound [K+].[K+].[O-]C([O-])=O BWHMMNNQKKPAPP-UHFFFAOYSA-L 0.000 claims abstract description 11
- 229910000019 calcium carbonate Inorganic materials 0.000 claims abstract description 10
- 229910052799 carbon Inorganic materials 0.000 claims abstract description 10
- -1 glycerol ester Chemical class 0.000 claims abstract description 10
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 claims abstract description 9
- ZLNQQNXFFQJAID-UHFFFAOYSA-L magnesium carbonate Chemical compound [Mg+2].[O-]C([O-])=O ZLNQQNXFFQJAID-UHFFFAOYSA-L 0.000 claims abstract description 7
- AXCZMVOFGPJBDE-UHFFFAOYSA-L calcium dihydroxide Chemical compound [OH-].[OH-].[Ca+2] AXCZMVOFGPJBDE-UHFFFAOYSA-L 0.000 claims abstract description 6
- 239000001095 magnesium carbonate Substances 0.000 claims abstract description 6
- 229910000021 magnesium carbonate Inorganic materials 0.000 claims abstract description 6
- 239000000920 calcium hydroxide Substances 0.000 claims abstract description 5
- 229910001861 calcium hydroxide Inorganic materials 0.000 claims abstract description 5
- 229910000027 potassium carbonate Inorganic materials 0.000 claims abstract description 5
- 239000000654 additive Substances 0.000 claims abstract description 3
- 239000003921 oil Substances 0.000 claims description 64
- 239000003925 fat Substances 0.000 claims description 59
- 235000019197 fats Nutrition 0.000 claims description 59
- 238000000034 method Methods 0.000 abstract description 17
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerol Natural products OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 abstract description 11
- 239000001963 growth medium Substances 0.000 abstract 4
- 235000019198 oils Nutrition 0.000 description 58
- 239000000203 mixture Substances 0.000 description 14
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 description 12
- VLKZOEOYAKHREP-UHFFFAOYSA-N n-Hexane Chemical compound CCCCCC VLKZOEOYAKHREP-UHFFFAOYSA-N 0.000 description 9
- 229940041514 candida albicans extract Drugs 0.000 description 8
- 244000005700 microbiome Species 0.000 description 8
- 235000020777 polyunsaturated fatty acids Nutrition 0.000 description 8
- 239000012138 yeast extract Substances 0.000 description 8
- 230000000052 comparative effect Effects 0.000 description 7
- 235000014113 dietary fatty acids Nutrition 0.000 description 7
- 229930195729 fatty acid Natural products 0.000 description 7
- 239000000194 fatty acid Substances 0.000 description 7
- 235000021323 fish oil Nutrition 0.000 description 7
- 241000894006 Bacteria Species 0.000 description 6
- 150000004665 fatty acids Chemical group 0.000 description 6
- 239000007788 liquid Substances 0.000 description 6
- 229910052943 magnesium sulfate Inorganic materials 0.000 description 6
- 235000019341 magnesium sulphate Nutrition 0.000 description 6
- 238000011084 recovery Methods 0.000 description 6
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 5
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 description 5
- 229910052921 ammonium sulfate Inorganic materials 0.000 description 5
- 235000011130 ammonium sulphate Nutrition 0.000 description 5
- 238000000605 extraction Methods 0.000 description 5
- 235000011187 glycerol Nutrition 0.000 description 5
- IPCSVZSSVZVIGE-UHFFFAOYSA-N hexadecanoic acid Chemical compound CCCCCCCCCCCCCCCC(O)=O IPCSVZSSVZVIGE-UHFFFAOYSA-N 0.000 description 4
- 239000008363 phosphate buffer Substances 0.000 description 4
- 241000251468 Actinopterygii Species 0.000 description 3
- 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 3
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 3
- 230000015572 biosynthetic process Effects 0.000 description 3
- 230000008859 change Effects 0.000 description 3
- 235000019688 fish Nutrition 0.000 description 3
- 239000008103 glucose Substances 0.000 description 3
- 235000001727 glucose Nutrition 0.000 description 3
- 239000002689 soil Substances 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- 241000222120 Candida <Saccharomycetales> Species 0.000 description 2
- 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 2
- 102000004882 Lipase Human genes 0.000 description 2
- 108090001060 Lipase Proteins 0.000 description 2
- 239000004367 Lipase Substances 0.000 description 2
- 235000021314 Palmitic acid Nutrition 0.000 description 2
- 229920002472 Starch Polymers 0.000 description 2
- 235000021355 Stearic acid Nutrition 0.000 description 2
- XSQUKJJJFZCRTK-UHFFFAOYSA-N Urea Chemical compound NC(N)=O XSQUKJJJFZCRTK-UHFFFAOYSA-N 0.000 description 2
- 230000034303 cell budding Effects 0.000 description 2
- 239000012141 concentrate Substances 0.000 description 2
- 238000007796 conventional method Methods 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 125000005456 glyceride group Chemical group 0.000 description 2
- 229910017053 inorganic salt Inorganic materials 0.000 description 2
- 235000019421 lipase Nutrition 0.000 description 2
- 125000002496 methyl group Chemical group [H]C([H])([H])* 0.000 description 2
- WQEPLUUGTLDZJY-UHFFFAOYSA-N n-Pentadecanoic acid Natural products CCCCCCCCCCCCCCC(O)=O WQEPLUUGTLDZJY-UHFFFAOYSA-N 0.000 description 2
- QIQXTHQIDYTFRH-UHFFFAOYSA-N octadecanoic acid Chemical compound CCCCCCCCCCCCCCCCCC(O)=O QIQXTHQIDYTFRH-UHFFFAOYSA-N 0.000 description 2
- OQCDKBAXFALNLD-UHFFFAOYSA-N octadecanoic acid Natural products CCCCCCCC(C)CCCCCCCCC(O)=O OQCDKBAXFALNLD-UHFFFAOYSA-N 0.000 description 2
- 238000000746 purification Methods 0.000 description 2
- 239000002994 raw material Substances 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 150000003839 salts Chemical class 0.000 description 2
- 235000003441 saturated fatty acids Nutrition 0.000 description 2
- 150000004671 saturated fatty acids Chemical class 0.000 description 2
- 238000012216 screening Methods 0.000 description 2
- 239000008107 starch Substances 0.000 description 2
- 235000019698 starch Nutrition 0.000 description 2
- 239000008117 stearic acid Substances 0.000 description 2
- 239000006228 supernatant Substances 0.000 description 2
- 239000008399 tap water Substances 0.000 description 2
- 235000020679 tap water Nutrition 0.000 description 2
- 238000012360 testing method Methods 0.000 description 2
- UFTFJSFQGQCHQW-UHFFFAOYSA-N triformin Chemical compound O=COCC(OC=O)COC=O UFTFJSFQGQCHQW-UHFFFAOYSA-N 0.000 description 2
- 235000021122 unsaturated fatty acids Nutrition 0.000 description 2
- 150000004670 unsaturated fatty acids Chemical class 0.000 description 2
- 235000013343 vitamin Nutrition 0.000 description 2
- 239000011782 vitamin Substances 0.000 description 2
- 229940088594 vitamin Drugs 0.000 description 2
- 229930003231 vitamin Natural products 0.000 description 2
- 150000003722 vitamin derivatives Chemical class 0.000 description 2
- HDTRYLNUVZCQOY-UHFFFAOYSA-N α-D-glucopyranosyl-α-D-glucopyranoside Natural products OC1C(O)C(O)C(CO)OC1OC1C(O)C(O)C(O)C(CO)O1 HDTRYLNUVZCQOY-UHFFFAOYSA-N 0.000 description 1
- CDVZCUKHEYPEQS-SZBOBFKXSA-N (2R,3R,4R)-2,3,4,5-tetrahydroxypentanal (2R,3S,4S)-2,3,4,5-tetrahydroxypentanal Chemical compound OC[C@H](O)[C@H](O)[C@@H](O)C=O.OC[C@@H](O)[C@@H](O)[C@@H](O)C=O CDVZCUKHEYPEQS-SZBOBFKXSA-N 0.000 description 1
- WRIDQFICGBMAFQ-UHFFFAOYSA-N (E)-8-Octadecenoic acid Natural products CCCCCCCCCC=CCCCCCCC(O)=O WRIDQFICGBMAFQ-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
- LQJBNNIYVWPHFW-UHFFFAOYSA-N 20:1omega9c fatty acid Natural products CCCCCCCCCCC=CCCCCCCCC(O)=O LQJBNNIYVWPHFW-UHFFFAOYSA-N 0.000 description 1
- QSBYPNXLFMSGKH-UHFFFAOYSA-N 9-Heptadecensaeure Natural products CCCCCCCC=CCCCCCCCC(O)=O QSBYPNXLFMSGKH-UHFFFAOYSA-N 0.000 description 1
- 229920001817 Agar Polymers 0.000 description 1
- KRKNYBCHXYNGOX-UHFFFAOYSA-K Citrate Chemical compound [O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O KRKNYBCHXYNGOX-UHFFFAOYSA-K 0.000 description 1
- 241000555825 Clupeidae Species 0.000 description 1
- 241000238424 Crustacea Species 0.000 description 1
- 241000195493 Cryptophyta Species 0.000 description 1
- GUBGYTABKSRVRQ-CUHNMECISA-N D-Cellobiose Chemical compound 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-CUHNMECISA-N 0.000 description 1
- SHZGCJCMOBCMKK-UHFFFAOYSA-N D-mannomethylose Natural products CC1OC(O)C(O)C(O)C1O SHZGCJCMOBCMKK-UHFFFAOYSA-N 0.000 description 1
- SRBFZHDQGSBBOR-IOVATXLUSA-N D-xylopyranose Chemical compound O[C@@H]1COC(O)[C@H](O)[C@H]1O SRBFZHDQGSBBOR-IOVATXLUSA-N 0.000 description 1
- PNNNRSAQSRJVSB-UHFFFAOYSA-N L-rhamnose Natural products CC(O)C(O)C(O)C(O)C=O PNNNRSAQSRJVSB-UHFFFAOYSA-N 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
- 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
- 241001465754 Metazoa Species 0.000 description 1
- 229910002651 NO3 Inorganic materials 0.000 description 1
- NHNBFGGVMKEFGY-UHFFFAOYSA-N Nitrate Chemical compound [O-][N+]([O-])=O NHNBFGGVMKEFGY-UHFFFAOYSA-N 0.000 description 1
- 239000005642 Oleic acid Substances 0.000 description 1
- ZQPPMHVWECSIRJ-UHFFFAOYSA-N Oleic acid Natural products CCCCCCCCC=CCCCCCCCC(O)=O ZQPPMHVWECSIRJ-UHFFFAOYSA-N 0.000 description 1
- 229910019142 PO4 Inorganic materials 0.000 description 1
- 241000269851 Sarda sarda Species 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
- HDTRYLNUVZCQOY-WSWWMNSNSA-N Trehalose Natural products O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@H](O)[C@@H](O)[C@@H](O)[C@@H](CO)O1 HDTRYLNUVZCQOY-WSWWMNSNSA-N 0.000 description 1
- 238000009825 accumulation Methods 0.000 description 1
- 238000005273 aeration Methods 0.000 description 1
- 239000008272 agar Substances 0.000 description 1
- PNNNRSAQSRJVSB-BXKVDMCESA-N aldehydo-L-rhamnose Chemical compound C[C@H](O)[C@H](O)[C@@H](O)[C@@H](O)C=O PNNNRSAQSRJVSB-BXKVDMCESA-N 0.000 description 1
- HDTRYLNUVZCQOY-LIZSDCNHSA-N alpha,alpha-trehalose Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 HDTRYLNUVZCQOY-LIZSDCNHSA-N 0.000 description 1
- 150000003863 ammonium salts Chemical class 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 230000001580 bacterial effect Effects 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- SRBFZHDQGSBBOR-UHFFFAOYSA-N beta-D-Pyranose-Lyxose Natural products OC1COC(O)C(O)C1O SRBFZHDQGSBBOR-UHFFFAOYSA-N 0.000 description 1
- 159000000007 calcium salts Chemical class 0.000 description 1
- 239000004202 carbamide Substances 0.000 description 1
- 150000001720 carbohydrates Chemical class 0.000 description 1
- 150000001721 carbon Chemical group 0.000 description 1
- 125000004432 carbon atom Chemical group C* 0.000 description 1
- 239000003054 catalyst Substances 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 238000004587 chromatography analysis Methods 0.000 description 1
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 description 1
- 150000001879 copper Chemical class 0.000 description 1
- 238000002425 crystallisation Methods 0.000 description 1
- 239000012228 culture supernatant Substances 0.000 description 1
- 230000002950 deficient Effects 0.000 description 1
- 239000012954 diazonium Substances 0.000 description 1
- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000010701 ester synthesis reaction Methods 0.000 description 1
- 229940013317 fish oils Drugs 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 238000005194 fractionation Methods 0.000 description 1
- 235000021588 free fatty acids Nutrition 0.000 description 1
- 229930182830 galactose Natural products 0.000 description 1
- 159000000014 iron salts Chemical class 0.000 description 1
- QXJSBBXBKPUZAA-UHFFFAOYSA-N isooleic acid Natural products CCCCCCCC=CCCCCCCCCC(O)=O QXJSBBXBKPUZAA-UHFFFAOYSA-N 0.000 description 1
- 235000020778 linoleic acid Nutrition 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
- 230000037356 lipid metabolism Effects 0.000 description 1
- 159000000003 magnesium salts Chemical class 0.000 description 1
- 235000010355 mannitol Nutrition 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 230000000813 microbial effect Effects 0.000 description 1
- 239000011785 micronutrient Substances 0.000 description 1
- 235000013369 micronutrients Nutrition 0.000 description 1
- 235000013379 molasses Nutrition 0.000 description 1
- 230000000877 morphologic effect Effects 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- QJGQUHMNIGDVPM-UHFFFAOYSA-N nitrogen group Chemical group [N] QJGQUHMNIGDVPM-UHFFFAOYSA-N 0.000 description 1
- ZQPPMHVWECSIRJ-KTKRTIGZSA-N oleic acid Chemical compound CCCCCCCC\C=C/CCCCCCCC(O)=O ZQPPMHVWECSIRJ-KTKRTIGZSA-N 0.000 description 1
- 235000021317 phosphate Nutrition 0.000 description 1
- 150000003013 phosphoric acid derivatives Chemical class 0.000 description 1
- 230000001766 physiological effect Effects 0.000 description 1
- 239000008057 potassium phosphate buffer Substances 0.000 description 1
- 159000000001 potassium salts Chemical class 0.000 description 1
- 238000001556 precipitation Methods 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 235000019512 sardine Nutrition 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 230000002269 spontaneous effect Effects 0.000 description 1
- 238000003756 stirring Methods 0.000 description 1
- KDYFGRWQOYBRFD-UHFFFAOYSA-L succinate(2-) Chemical compound [O-]C(=O)CCC([O-])=O KDYFGRWQOYBRFD-UHFFFAOYSA-L 0.000 description 1
- 239000005720 sucrose Substances 0.000 description 1
Landscapes
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Fats And Perfumes (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
Description
【0001】[0001]
【産業上の利用分野】この発明は、食品または医薬品等
として有用なドコサヘキサエン酸高含有油脂の製造方法
に関し、さらに詳しくは微生物によるドコサヘキサエン
酸グリセリンエステルの含有量を高めたドコサヘキサエ
ン酸高含有油脂の製造方法に関する。BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a method for producing fats and oils having a high content of docosahexaenoic acid, which are useful as foods or pharmaceuticals. Regarding the method.
【0002】[0002]
【従来の技術】ドコサヘキサエン酸(以下、DHAと略
記する)は、長鎖高度不飽和脂肪酸(以下PUFAと略
記する)の一種であり、多くの生理活性機能を持つこと
が知られたものであって、近年、濃縮や高度精製技術に
関する研究が活発に進められている。Docosahexaenoic acid (hereinafter abbreviated as DHA) is a kind of long-chain highly unsaturated fatty acid (hereinafter abbreviated as PUFA), and is known to have many physiologically active functions. In recent years, research on concentration and advanced purification technology has been actively promoted.
【0003】従来のPUFAまたはPUFA含有油脂の
濃縮方法としては、魚から抽出した油脂を低温分別結晶
法、尿素付加法、クロマト法、リパーゼ処理法等により
分離濃縮する方法が知られている。As a conventional method for concentrating PUFA or PUFA-containing fats and oils, a method is known in which fats and oils extracted from fish are separated and concentrated by a low temperature fractionation crystallization method, a urea addition method, a chromatography method, a lipase treatment method and the like.
【0004】[0004]
【発明が解決しようとする課題】しかし、上記した従来
のPUFAまたはPUFA含有油脂の濃縮方法では、煩
雑な操作を必要としたり、操作の煩雑性に伴う高コスト
の問題点があり、さらには、PUFAをグリセリンエス
テルの形で得る場合に、濃縮後、さらに別工程を加えな
ければならないという問題点もある。However, the above-mentioned conventional method for concentrating PUFA or PUFA-containing fats and oils involves complicated operations, and there is a problem of high cost due to the complexity of operations. When PUFA is obtained in the form of glycerin ester, there is also a problem that another step must be added after concentration.
【0005】また、微生物を用いたPUFA含有油脂の
製造方法としては、トルロプシス属に属する酵母を用い
たPUFAグリセリドの製造方法(特開平2−1698
8)が知られているが、この酵母はDHAのみ特異的に
資化能を欠如しているとはいい難いので、DHAの濃縮
効率の点で充分に改良された方法とはいえない。As a method for producing a PUFA-containing fat or oil using a microorganism, a method for producing a PUFA glyceride using yeast belonging to the genus Torulopsis (Japanese Patent Laid-Open No. 2-1698).
8) is known, but it is difficult to say that this yeast lacks the assimilation ability specifically only for DHA, and thus it cannot be said that it is a sufficiently improved method in terms of DHA concentration efficiency.
【0006】なお、魚から抽出、精製される以外の方法
として、微生物により生産されるDHAを含む油脂を抽
出する方法もあるが、培養単位容量当りのDHAの収率
はまだまだ低く、実用化にはコスト面での問題がある。As a method other than extraction and purification from fish, there is a method of extracting fats and oils containing DHA produced by microorganisms, but the yield of DHA per unit volume of culture is still low, and it is practically used. Has a cost problem.
【0007】そこで、この発明の課題は、上記した問題
点を解決して、DHAをグリセリンエステルの形で高濃
度に含有する油脂を、簡便な手法で効率よく製造できる
ようにすることである。[0007] Therefore, an object of the present invention is to solve the above-mentioned problems and to efficiently produce a fat or oil containing DHA in a high concentration in the form of glycerin ester by a simple method.
【0008】[0008]
【課題を解決するための手段】本願の発明者は、濃縮さ
れたDHA含有油脂を効率よく得る方法について検討を
重ねた結果、DHAを含有する油脂を炭素源とする培地
において、特定の微生物、即ちキャンディダ・ファマタ
US−238を培養することにより、油脂中のDHA
含有量を飛躍的に効率よく高められることを見いだし、
この発明を完成するに至った。Means for Solving the Problems As a result of extensive studies on a method for efficiently obtaining a concentrated DHA-containing fat or oil, the inventor of the present application has shown that in a medium containing a DHA-containing fat or oil as a carbon source, a specific microorganism, That is, by culturing Candida famata US-238,
We found that the content can be increased dramatically and efficiently,
The present invention has been completed.
【0009】すなわち、前記の課題を解決するため、こ
の発明においては、キャンディダ・ファマタ US−2
38(FERM P−13974)をドコサヘキサエン
酸含有油脂を炭素源とする培地に培養し、この培地中の
ドコサヘキサエン酸グリセリンエステルの濃度を高めて
油脂分を回収したのである。In other words, in order to solve the above-mentioned problems, according to the present invention, Candida Famata US-2 is used.
No. 38 (FERM P-13974) was cultured in a medium containing a docosahexaenoic acid-containing oil or fat as a carbon source, and the concentration of docosahexaenoic acid glycerin ester in this medium was increased to recover the oil or fat.
【0010】また、前記手段において、キャンディダ・
ファマタ US−238(FERMP−13974)を
培養する培地が、ドコサヘキサエン酸を5重量%以上含
有する油脂を炭素源とする培地を採用することができ
る。Further, in the above-mentioned means,
As a medium for culturing Famata US-238 (FERMP-13974), a medium containing an oil or fat containing 5% by weight or more of docosahexaenoic acid as a carbon source can be adopted.
【0011】または、上記したドコサヘキサエン酸高含
有油脂の製造方法において、培地に炭酸カルシウム、水
酸化カルシウム、炭酸カリウムおよび炭酸マグネシウム
からなる群から選ばれる一種以上の添加剤を0.1〜1
容量(w/V)%添加する手段を採用することもでき
る。Alternatively, in the above-mentioned method for producing fats and oils having a high content of docosahexaenoic acid, 0.1 to 1 of one or more additives selected from the group consisting of calcium carbonate, calcium hydroxide, potassium carbonate and magnesium carbonate is added to the medium.
It is also possible to adopt a means of adding the volume (w / V)%.
【0012】以下に、その詳細を述べる。なお、この発
明で用いる%は、特にことわりのない限り、容量(v/
W)%で示した。この発明に用いるキャンディダ・ファ
マタ US−238(Candidafamata U
S−238)は、本願の発明者が日本各地で採取された
土壌から微生物を分離して、その油脂に対する資化能を
調べた結果、本願発明の目的を最も効率よく達成できる
ものとして採用された菌株であり、これは広島県因島の
土壌から分離し、次のスクリーニング方法により単離し
たものである。The details will be described below. The% used in the present invention is the capacity (v /
W)%. Candida famata U-238 (Candida famata U used for this invention)
S-238) was adopted by the inventor of the present application as the one that most efficiently achieves the object of the present invention as a result of separating microorganisms from soils collected in various parts of Japan and examining their assimilation ability for fats and oils. This strain was isolated from the soil of Innoshima Island, Hiroshima Prefecture, and was isolated by the following screening method.
【0013】[キャンディダ・ファマタ US−238
のスクリーニング方法]土壌の分離サンプルを下記組成
の培地Aに添加し、30℃で2〜3日培養した後、微生
物の増殖が見られるものについて、同じ培地で集積培養
を行ない、油脂資化能の強い菌を集積させた。そして、
培養液を遠心分離処理することによって上層にある油脂
の減少量を確認し、最も油脂資化能の強い菌を含む培養
液を同様な組成で構成した平板培地上に塗布し、培養
後、コロニーを単離し分取して目的とする菌株を得た。[Candida Famata US-238
Screening method] The soil separation sample was added to the medium A having the following composition, and the mixture was cultured at 30 ° C. for 2 to 3 days. Then, for those in which the growth of microorganisms was observed, the accumulation culture was performed in the same medium to obtain the oil and fat assimilation ability. Accumulated strong bacteria. And
Confirm the reduction amount of oil and fat in the upper layer by centrifuging the culture solution, apply the culture solution containing the bacteria with the highest oil and fat assimilation ability on a plate medium composed of the same composition, after culturing, colonies Was isolated and fractionated to obtain the target strain.
【0014】培地Aの組成:(pH8.0) 魚油 1 ml グルコース 3.5 mg 硝酸アンモニウム 35 mg リン酸水素二カリウム 14 mg 硫酸マグネシウム 7 mg 酵母エキス 7 mg 水道水 7 ml 前記得られた菌株の菌学的性状は以下の如くである。Composition of medium A: (pH 8.0) Fish oil 1 ml Glucose 3.5 mg Ammonium nitrate 35 mg Dipotassium hydrogen phosphate 14 mg Magnesium sulfate 7 mg Yeast extract 7 mg Tap water 7 ml Bacteria of the strain obtained above The scientific properties are as follows.
【0015】(1) 形態的性質 形態 卵型又は球形 細胞は1個又は数個連な
っている 大きさ (1〜6)×(2〜6)μ 出芽 多極出芽 偽菌糸 コーンミール培地で未発達な偽菌糸の形
成を認める 子嚢胞子 形成を認めず (2) 培養的性質(YM液体培地) 沈澱及び皮膜の形成を認める (3) 巨大コロニーの観察(YM寒天培地) 成育 良好 ***状態 台状 表面の状態 平滑 色調 乳白色 (4) 生理学的性質 生育pH 3〜9 最適pH 5〜7 生育温度 約42℃まで 最適温度 20〜35 硝酸塩の資化性 − ビタミン欠培地での生育 − ジアゾニウムブルーBの呈色反応 − [発酵性] グルコース − ガラクトース − シュークロース − マルトース − ラクトース − ラフィノース − イノシトール − [資化性] グルコース + ガラクトース + シュークロース + マルトース + セロビオース + トレハロース + ラクトース − ラフィノース + メリチトース + スターチ + D−キシロース + L−アラビノース − D−リボース + L−ラムノース + エリスリトール − リビトール + D−マンニトール + コハク酸塩 + クエン酸塩 + イノシトール − 以上の結果から、分離された菌株は、キャンディダ・フ
ァマタ(Candida famata)であると同定
され、この株をキャンディダ・ファマタ US−238
(Candida famata US−238)と命
名した。(1) Morphological properties Morphology Oval or globular cells are one or several in series. Size (1-6) × (2-6) μ Budding multipolar budding Pseudohyphae Not developed in cornmeal medium Spontaneous mycelium formation is observed No ascospore formation is observed (2) Cultural properties (YM liquid medium) Precipitation and film formation are observed (3) Giant colony observation (YM agar medium) Good growth Upstanding trapezoid Surface condition Smooth tone Milky white (4) Physiological properties Growth pH 3-9 Optimum pH 5-7 Growth temperature Up to about 42 ℃ Optimum temperature 20-35 Nitrate assimilation-Growth in vitamin-deficient medium-Diazonium blue B Color reaction- [fermentability] glucose-galactose-sucrose-maltose-lactose-raffinose-inositol- [assimilation] glucose + galactose + sucrose + Maltose + cellobiose + trehalose + lactose-raffinose + melitithose + starch + D-xylose + L-arabinose-D-ribose + L-rhamnose + erythritol-ribitol + D-mannitol + succinate + citrate- + citrate. From the results, the isolated strain was identified as Candida famata, and this strain was designated as Candida famata US-238.
(Candida famata US-238).
【0016】この菌株は、工業技術院生命工学工業技術
研究所に「微生物受託番号 生命研菌寄第13974号
(FERM P−13974)」として寄託されてい
る。This strain has been deposited at the Institute of Biotechnology, Institute of Biotechnology, Institute of Industrial Science and Technology as "Microbial Accession No. Life Research Institute No. 13974 (FERM P-13974)".
【0017】この発明におけるDHAは、炭素数22で
6個の二重結合を持っており、メチル基側から数えて3
番目の炭素原子から二重結合が始まる脂肪酸である。こ
のようなDHAを含有する油脂としては、カツオ、マグ
ロ、イワシ等の魚類の油(魚油)や、甲殻類、海産動物
類、藻類等の油脂を挙げることができる。The DHA used in the present invention has 6 double bonds with 22 carbon atoms, and has 3 double bonds counted from the methyl group side.
It is a fatty acid in which the double bond begins at the th carbon atom. Examples of such oils and fats containing DHA include oils (fish oils) of fish such as bonito, tuna and sardines, and oils and fats of crustaceans, marine animals, algae and the like.
【0018】この発明における培養は、DHAを含有す
る油脂を炭素源として含む培地に前記の酵母を接種し、
好気的に培養することにより行なわれるが、その場合に
用いる炭素源としては、DHA含有油脂のほか、グルコ
ース、デンプン、廃糖蜜等の糖類を併用することができ
る。In the culture according to the present invention, the above yeast is inoculated into a medium containing a fat and oil containing DHA as a carbon source,
It is carried out by aerobically culturing, and as the carbon source used in that case, saccharides such as glucose, starch, molasses and the like can be used in addition to the DHA-containing oil and fat.
【0019】また、窒素源としてはアンモニウム塩、そ
の他の窒素含有物質を用いることができ、無機塩類とし
ては、マグネシウム塩、リン酸塩、カリウム塩、カルシ
ウム塩、鉄塩、銅塩等を用いることができる。また、ビ
タミン源としては、酵母エキスが用いられる他、成長促
進物質を添加することも好ましい。Ammonium salts and other nitrogen-containing substances can be used as the nitrogen source, and magnesium salts, phosphates, potassium salts, calcium salts, iron salts, copper salts, etc. can be used as the inorganic salts. You can In addition to yeast extract, it is also preferable to add a growth promoting substance as a vitamin source.
【0020】この発明に用いる培地の炭素源は、DHA
を5重量%以上含有する油脂を用いることが好ましい。
この発明では、DHA以外の成分を特定の酵母で資化す
ることによって油脂中のDHAを濃縮するのであるか
ら、DHA含有量が5重量%未満の油脂では、製造効率
が悪くなるからである。The carbon source of the medium used in this invention is DHA.
It is preferable to use fats and oils containing 5% by weight or more.
This is because, in the present invention, the components other than DHA are assimilated by the specific yeast to concentrate the DHA in the fats and oils, and thus the fats and oils having a DHA content of less than 5% by weight deteriorate the production efficiency.
【0021】培地における初発のDHA含有油脂濃度
は、500g/1以下で行なうが、より好ましい濃度範
囲は10〜300g/1である。初発量に応じて培養時
間が長くなり、300g/1を越えると実用性が低くな
る。The initial concentration of DHA-containing fats and oils in the medium is 500 g / 1 or less, and a more preferable concentration range is 10 to 300 g / 1. Cultivation time becomes longer depending on the initial amount, and when it exceeds 300 g / 1, the practicality becomes low.
【0022】培養温度は20〜40℃、好ましくは25
〜35℃で、pHは3〜9、好ましくは5〜7にて通常
3〜10日行えばよい。The culture temperature is 20 to 40 ° C., preferably 25.
The pH is 3 to 9, preferably 5 to 7 at ˜35 ° C., and usually 3 to 10 days.
【0023】また無機塩類は、微生物の微量栄養素とし
て、通常は0.01〜0.5%の範囲で用いられること
が多いが、この発明においては、培養途中例えば培養2
〜5日後に炭酸カルシウム、水酸化カルシウム、炭酸カ
リウム、炭酸マグネシウムの中から選ばれる1種、また
はそれらの混合物を0.1〜1%添加することによりさ
らにDHA含量を高めることが可能である。すなわち、
この発明に用いる微生物は、通常の培養時において特に
オレイン酸、リノール酸等不飽和脂肪酸の資化能に優れ
ている。Inorganic salts are often used in the range of 0.01 to 0.5% as a micronutrient for microorganisms, but in the present invention, during the culture, for example, culture 2 is used.
After 5 days, the DHA content can be further increased by adding 0.1 to 1% of one selected from calcium carbonate, calcium hydroxide, potassium carbonate, magnesium carbonate, or a mixture thereof. That is,
The microorganism used in the present invention is particularly excellent in assimilation ability of unsaturated fatty acids such as oleic acid and linoleic acid during ordinary culture.
【0024】しかし、上記無機塩を添加することにより
パルミチン酸、ステアリン酸等の飽和脂肪酸の資化能が
増し、結果的にDHA含量がさらに高まることになる。
上記無機塩添加による微生物の資化能の変化の理由につ
いては明らかではないが、脂質代謝機構に何らかの変化
を与えているものと考えられる。但し、添加量が1.0
%をこえると濃縮を阻害する傾向が現れる。However, the addition of the above-mentioned inorganic salt increases the assimilation ability of saturated fatty acids such as palmitic acid and stearic acid, resulting in a further increase in the DHA content.
Although the reason for the change in the assimilation ability of the microorganism due to the addition of the inorganic salt is not clear, it is considered that it causes some change in the lipid metabolism mechanism. However, the addition amount is 1.0
If it exceeds%, the concentration tends to be inhibited.
【0025】DHA含有油脂の回収方法としては、培養
終了後、菌体を遠心分離等によって除き、得られた培養
上清からヘキサン等の油脂抽出溶剤を用いて、DHA含
有油脂を抽出回収すればよく、濃縮されたDHA含有油
脂を得ることができる。As a method for recovering the DHA-containing oil / fat, after the culture is completed, the cells are removed by centrifugation or the like, and the DHA-containing oil / fat is extracted and recovered from the obtained culture supernatant using an oil / fat extraction solvent such as hexane. Well, concentrated DHA-containing fats and oils can be obtained.
【0026】なお、得られたDHA高含有性油脂中のD
HAは、各種グリセリン脂肪酸エステルの混合物、特に
トリグリセリドとジグリセリドの混合物として得られ
る。必要ならばこのグリセリド混合物を適当な触媒、例
えばリパーゼによりエステル合成反応を行えばDHA高
含有性のトリグリセリドが得られる。D in the obtained DHA-rich oil and fat
HA is obtained as a mixture of various glycerin fatty acid esters, particularly a mixture of triglyceride and diglyceride. If necessary, this glyceride mixture is subjected to an ester synthesis reaction with a suitable catalyst such as lipase to obtain a triglyceride having a high DHA content.
【0027】[0027]
【実施例】次に実施例および比較例によりこの発明をさ
らに詳しく説明する。 〔実施例1〕硫酸アンモニウム0.5%、硫酸マグネシ
ウム0.1%、酵母エキス0.1%を含み、0.25M
リン酸緩衝液でpH6に調製された液体培地100ml
を坂口フラスコに入れ、さらにDHA含有油脂として魚
油を5.0g添加し、キャンディダ・ファマタ US−
238を接種し、30℃で3日間振とう培養した後、培
養液を遠心分離して菌体を除いた上清をヘキサンで抽出
し油分を回収した。得られた油脂の脂肪酸組成を表1に
示した。EXAMPLES The present invention will be described in more detail with reference to Examples and Comparative Examples. [Example 1] 0.25 M containing ammonium sulfate 0.5%, magnesium sulfate 0.1%, yeast extract 0.1%
100 ml of liquid medium adjusted to pH 6 with phosphate buffer
Was added to a Sakaguchi flask and 5.0 g of fish oil was added as a DHA-containing oil and fat. Candida Famata US-
After inoculating 238 with shaking and culturing at 30 ° C. for 3 days with shaking, the culture solution was centrifuged to remove the cells, and the supernatant was extracted with hexane to recover an oil component. The fatty acid composition of the obtained fats and oils is shown in Table 1.
【0028】なお、脂肪酸組成分析は、油脂をメチルエ
ステル化し、GCにて測定し、全組成から要部を抜粋し
た。In the fatty acid composition analysis, fats and oils were methyl esterified, measured by GC, and essential parts were extracted from all compositions.
【0029】[0029]
【表1】 [Table 1]
【0030】表1の結果からも明らかなように、原料油
脂のDHA含有量は19.5%であったのに対し、キャ
ンディダ・ファマタ US−238を培養処理後のDH
A含有量は35.2%であった。したがってDHAは約
1.8倍に濃縮されたことになる。なお油脂の回収率は
45%であった。As is clear from the results shown in Table 1, the DHA content of the raw material fat was 19.5%, whereas the DH after the Candida famata US-238 was subjected to the culture treatment.
The A content was 35.2%. Therefore, DHA is concentrated about 1.8 times. The oil and fat recovery rate was 45%.
【0031】〔実施例2〕実施例1において培養開始時
に炭酸カルシウム0.5%を添加して培養を行なったこ
と以外は全く同様にして、培養後、抽出を行ない、油脂
の脂肪酸組成及びDHA含量を調べ、結果を表1中に併
記した。[Example 2] [0031] Extraction was carried out in the same manner as in Example 1 except that 0.5% of calcium carbonate was added at the start of the culture, and then extraction was carried out to obtain the fatty acid composition of fats and oils and DHA. The content was investigated and the results are also shown in Table 1.
【0032】その結果、炭酸カルシウムを添加すること
により、本発明に使用する菌の資化によって残存する脂
肪酸に変化がみられた。すなわち、パルミチン酸、ステ
アリン酸等飽和脂肪酸の資化能が増し、結果的にDHA
含量が高まっていた。As a result, the addition of calcium carbonate caused a change in the residual fatty acid due to the assimilation of the bacterium used in the present invention. That is, the assimilation ability of saturated fatty acids such as palmitic acid and stearic acid is increased, resulting in DHA.
The content was increasing.
【0033】〔実施例3〜7〕硫酸アンモニウム0.5
%、硫酸マグネシウム0.1%、酵母エキス0.1%を
含み、0.25Mリン酸緩衝液でpH6に調製された液
体培地7mlを試験管に入れ、さらにDHA含有油脂と
して魚油(DHA 19.5%)を0.7g添加し、キ
ャンディダ・ファマタ US−238を接種し、30℃
で2日間振とう培養した。[Examples 3 to 7] Ammonium sulfate 0.5
%, Magnesium sulfate 0.1%, yeast extract 0.1%, 7 ml of a liquid medium adjusted to pH 6 with a 0.25 M phosphate buffer was placed in a test tube, and fish oil (DHA 19. (5%) 0.7 g, and inoculated with Candida famata US-238, 30 ° C
The cells were shaken and cultured for 2 days.
【0034】その後、なにも添加せずそのまま培養を継
続したもの(実施例3)、炭酸カルシウム(CaC
O3 )0.5%添加したもの(実施例4)、水酸化カル
シウム(Ca(OH)2 )0.5%添加したもの(実施
例5)、炭酸カリウム(K2 CO3 )0.5%添加した
もの(実施例6)、炭酸マグネシウム0.5%添加した
もの(実施例7)を別途調製し、再び継続して3日間培
養した。これらは培養後、実施例1と全く同様にして油
脂分を回収し、得られた油脂の脂肪酸組成を調べた。こ
の結果を表2に示した。After that, the culture was continued without adding anything (Example 3), calcium carbonate (CaC
O 3) have been added 0.5% (Example 4), calcium hydroxide (Ca (OH) 2) which was added 0.5% (Example 5), potassium carbonate (K 2 CO 3) 0.5 % (Example 6) and 0.5% magnesium carbonate (Example 7) were separately prepared and continuously cultured for 3 days. After culturing, the fats and oils were collected in exactly the same manner as in Example 1, and the fatty acid composition of the obtained fats and oils was examined. The results are shown in Table 2.
【0035】[0035]
【表2】 [Table 2]
【0036】表2の結果からも明らかなように、炭酸カ
ルシウム、水酸化カルシウム、炭酸カリウムまたは炭酸
マグネシウムを培養途中に添加することによって、実施
例1、2に比べてさらにDHA含量が高まり、DHA含
有成分がさらに濃縮された油脂が得られたことがわか
る。As is clear from the results shown in Table 2, the addition of calcium carbonate, calcium hydroxide, potassium carbonate or magnesium carbonate during the culture further increased the DHA content as compared with Examples 1 and 2, and It can be seen that a fat or oil in which the contained components were further concentrated was obtained.
【0037】〔実施例8〜11〕硫酸アンモニウム0.
5%、硫酸マグネシウム0.1%、酵母エキス0.1%
を含み、0.25Mリン酸緩衝液でpH6に調製された
液体培地7mlを試験管に入れ、さらにDHA含有油脂
として魚油(DHA 26.8%)を0.7g添加し、
キャンディダ・ファマタ US−238を接種し、30
℃で3日間振とう培養した。その後、炭酸カルシウム
(CaCO3 をそれぞれ0.1%、0・2%0.5%、
1.0%添加し(それぞれ実施例8、9、10、1
1)、さらに継続して4日間培養した後、実施例1と全
く同様に油脂分を回収し、得られた油脂のDHA含有量
及び油脂の回収率を調べた。この結果を表3に示した。[Examples 8 to 11] Ammonium sulfate 0.
5%, magnesium sulfate 0.1%, yeast extract 0.1%
Into a test tube, 7 ml of a liquid medium containing, and adjusted to pH 6 with a 0.25 M phosphate buffer was added, and 0.7 g of fish oil (DHA 26.8%) as a DHA-containing oil was added.
Inoculated with Candida famata US-238, 30
The cells were shaken at 3 ° C for 3 days. After that, calcium carbonate (CaCO 3 0.1%, 0.2% 0.5%,
1.0% was added (Examples 8, 9, 10, 1 respectively)
1), after further continuous culturing for 4 days, the oil and fat was recovered in exactly the same manner as in Example 1, and the DHA content and the oil and fat recovery rate of the obtained oil and fat were examined. The results are shown in Table 3.
【0038】[0038]
【表3】 [Table 3]
【0039】表3の結果からも明らかなように、炭酸カ
ルシウムを0.1〜1.0%添加することによりDHA
がさらに濃縮され(実施例8〜11)、DHA含有量の
高い油脂が回収されたことがわかる。As is clear from the results shown in Table 3, addition of calcium carbonate in an amount of 0.1 to 1.0% resulted in DHA.
Was further concentrated (Examples 8 to 11), and it was found that fats and oils having a high DHA content were recovered.
【0040】〔比較例1、2〕実施例8においてCaC
O3 を添加しない(比較例1)、または培養3日後にC
aCO3 を2.0%添加した(比較例2)こと以外は実
施例8と全く同様に行い、回収した油脂のDHA含量及
び油脂回収率を調べ、結果を表3中に併記した。その結
果、2.0%では、無添加のものに比べ、DHA含量が
やや低くなっていた。[Comparative Examples 1 and 2] CaC in Example 8
O 3 was not added (Comparative Example 1), or C was added after 3 days of culture.
The same procedure as in Example 8 was carried out except that 2.0% of aCO 3 was added (Comparative Example 2), and the DHA content and the oil / fat recovery rate of the recovered oil / fat were examined, and the results are also shown in Table 3. As a result, at 2.0%, the DHA content was slightly lower than that of the non-added one.
【0041】〔実施例12〕硫酸アンモニウム0.5
%、硫酸マグネシウム0.1%、酵母エキス0.1%を
含み、0.25Mリン酸カリウム緩衝液でpH6に調製
された液体培地100mlを坂口フラスコに入れ、さら
にDHA含有魚油(DHA26.8%)を5g添加し、
キャンディダ・ファマタ US−238を接種し、30
℃で3日間振とう培養行い、実施例1と同様に油脂の抽
出操作を行い、DHA含量及び油脂回収率を調べ、この
結果を表4に示した。Example 12 Ammonium sulfate 0.5
%, Magnesium sulfate 0.1%, yeast extract 0.1%, 100 ml of a liquid medium adjusted to pH 6 with a 0.25 M potassium phosphate buffer was placed in a Sakaguchi flask, and DHA-containing fish oil (DHA 26.8% ) Is added,
Inoculated with Candida famata US-238, 30
The culture was carried out with shaking at 3 ° C. for 3 days, and the oil and fat extraction operation was performed in the same manner as in Example 1 to examine the DHA content and the oil and fat recovery rate. The results are shown in Table 4.
【0042】[0042]
【表4】 [Table 4]
【0043】〔比較例3、4〕実施例12においてキャ
ンディダ・ファマタ US−238株以外の株として、
Candida famata(IFO 0664)を
用い(比較例3)、または油脂資化性菌として知られて
いるTorulopsis属の菌としてTorulop
sis bombicola(IFO 10243)を
用いたこと(比較例4)以外は、全く同様に培養、油脂
の回収を行ない、結果を表4中に併記した。Comparative Examples 3 and 4 As strains other than the Candida famata US-238 strain in Example 12,
Candida famata (IFO 0664) was used (Comparative Example 3), or Torulop as a bacterium belonging to the genus Torulopsis known as an oil- and fat-assimilating bacterium.
Except that sis bombicola (IFO 10243) was used (Comparative Example 4), the culture and the oil / fat recovery were performed in exactly the same manner, and the results are also shown in Table 4.
【0044】表4の結果からも明らかなように、所定の
株以外を培養しても、回収された油脂中のDHA含有量
は31%以下であった。As is clear from the results shown in Table 4, the DHA content in the recovered oil was 31% or less even when the strains other than the predetermined strain were cultured.
【0045】〔実施例13〕硫酸アンモニウム0.5
%、硫酸マグネシウム0.1%、酵母エキス0.1%を
含み、0.25Mリン酸緩衝液でpH6に調製された液
体培地100mlを坂口フラスコに入れ、さらにDHA
含有油脂として魚油を5g添加し、キャンディダ・ファ
マタ US−238を接種し、30℃で6日間振とう培
養した。その後、CaO3を0.5%添加し、再び、継
続して3日間培養した後、培養液を遠心分離して菌体を
除いた上清をヘキサンで抽出した油分を回収した。[Example 13] Ammonium sulfate 0.5
%, Magnesium sulfate 0.1%, yeast extract 0.1%, 100 ml of liquid medium adjusted to pH 6 with 0.25M phosphate buffer was added to a Sakaguchi flask, and further DHA
5 g of fish oil was added as a contained oil and fat, and Candida famata US-238 was inoculated, followed by shaking culture at 30 ° C. for 6 days. After that, 0.5% of CaO3 was added, the culture was continued again for 3 days, and the culture solution was centrifuged to remove the bacterial cells, and the supernatant was extracted with hexane to recover an oil component.
【0046】この結果、原料油脂のDHA含有量は1
9.5%であったのに対し、酵母と接触処理後のDHA
含有量は、69.1%であった。したがってDHAは約
3.5倍に濃縮されたことになる。なお、油脂の回収率
は12.0%であった。As a result, the DHA content of the raw material oil is 1
9.5%, whereas DHA after contact treatment with yeast
The content was 69.1%. Therefore, DHA is concentrated about 3.5 times. The oil and fat recovery rate was 12.0%.
【0047】〔実施例14〕次に、ジャーファメンター
を用いて大量培養を行なった。培地の組成及び培養条件
は、下記に示した通りである。Example 14 Next, large-scale culture was carried out using a jar famenter. The composition of the medium and the culture conditions are as shown below.
【0048】[培地] 魚油(DHA 19.5%) 600g (NH4 )2 SO4 30g KH2 PO4 12g MgSO4 ・7H2 O 6g 酵母エキス 6g 水道水 6000ml [培養条件] pH 6.0(1Nの
NaOHで調節) 温度 30℃ 通気量 3 1/分 撹拌速度 300 rpm 培養時間 115 時間 CaCO3 30gの添加 培養開始67時間後 培養後、実施例1と全く同様に油脂を回収した結果、回
収油脂量は、115gであり、DHA含量は、37.1
%に濃縮されていた。[Medium] Fish oil (DHA 19.5%) 600 g (NH 4 ) 2 SO 4 30 g KH 2 PO 4 12 g MgSO 4 .7H 2 O 6 g Yeast extract 6 g Tap water 6000 ml [Cultivation conditions] pH 6.0 ( Adjusted with 1N NaOH) Temperature 30 ° C. Aeration rate 3 1 / min Stirring speed 300 rpm Culture time 115 hours Addition of CaCO 3 30 g 67 hours after the start of culture After the culture, the oil and fat were recovered in exactly the same manner as in Example 1, and recovered. The amount of oil and fat is 115 g, and the DHA content is 37.1.
It was concentrated to%.
【0049】[0049]
【効果】この発明は、以上説明したように、キャンディ
ダ・ファマタの所定菌株をDHA含有油脂用いた培地に
培養するといった簡易な方法により、DHAを含有する
油脂から効率よくDHAグリセリンエステルを濃縮回収
することができる利点がある。[Effect] As described above, the present invention efficiently concentrates and recovers DHA glycerin ester from DHA-containing fats and oils by a simple method such as culturing a predetermined strain of Candida famata in a medium containing DHA-containing fats and oils. There is an advantage that can be done.
【0050】また、培養の際に特定の無機物質を添加す
る手段を採用した発明では、さらにDHA含量を高める
ことができる。そして、この発明で得られるDHAは、
グリセリンエステルの形で存在し、遊離脂肪酸は、少量
しか存在しないため、培養後の油脂の精製が容易であ
り、非常に実用性の高い方法であるといえる。Further, in the invention which adopts a means for adding a specific inorganic substance at the time of culturing, the DHA content can be further increased. The DHA obtained by this invention is
Since it is present in the form of glycerin ester and the free fatty acid is present in a small amount, it can be said that the oil and fat after the culture are easily purified, which is a highly practical method.
Claims (3)
(FERM P−13974)をドコサヘキサエン酸含
有油脂を炭素源とする培地で培養し、この培地中のドコ
サヘキサエン酸グリセリンエステルの濃度を高めて油脂
分を回収することからなるドコサヘキサエン酸高含有油
脂の製造方法。1. Candida Famata US-238
(FERM P-13974) is cultured in a medium containing a docosahexaenoic acid-containing fat and oil as a carbon source, and the concentration of docosahexaenoic acid glycerin ester in the medium is increased to recover the fat and oil. .
(FERM P−13974)を培養する培地が、ドコ
サヘキサエン酸を5重量%以上含有する油脂を炭素源と
する培地である請求項1記載のドコサヘキサエン酸高含
有油脂の製造方法。2. Candida Famata US-238
The method for producing an oil and fat rich in docosahexaenoic acid according to claim 1, wherein the medium for culturing (FERM P-13974) is an oil and fat containing 5% by weight or more of docosahexaenoic acid as a carbon source.
エン酸高含有油脂の製造方法において、培地に炭酸カル
シウム、水酸化カルシウム、炭酸カリウムおよび炭酸マ
グネシウムからなる群から選ばれる一種以上の添加剤を
0.1〜1容量(w/V)%添加することを特徴とする
ドコサヘキサエン酸高含有油脂の製造方法。3. The method for producing a fat or oil rich in docosahexaenoic acid according to claim 1 or 2, wherein one or more additives selected from the group consisting of calcium carbonate, calcium hydroxide, potassium carbonate and magnesium carbonate are added to the medium. 0.1 to 1 volume (w / V)% is added, The manufacturing method of the fats and oils high in docosahexaenoic acid characterized by the above-mentioned.
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP6081654A JP2566377B2 (en) | 1994-04-20 | 1994-04-20 | Method for producing fats and oils high in docosahexaenoic acid |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP6081654A JP2566377B2 (en) | 1994-04-20 | 1994-04-20 | Method for producing fats and oils high in docosahexaenoic acid |
Publications (2)
Publication Number | Publication Date |
---|---|
JPH07289272A true JPH07289272A (en) | 1995-11-07 |
JP2566377B2 JP2566377B2 (en) | 1996-12-25 |
Family
ID=13752324
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP6081654A Expired - Fee Related JP2566377B2 (en) | 1994-04-20 | 1994-04-20 | Method for producing fats and oils high in docosahexaenoic acid |
Country Status (1)
Country | Link |
---|---|
JP (1) | JP2566377B2 (en) |
Cited By (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2006064317A1 (en) * | 2004-12-14 | 2006-06-22 | Avestha Gengraine Technologies Pvt. Ltd. | Recombinant production docosahexaenoic acid (dha) in yeast |
JP2009120840A (en) * | 1996-05-15 | 2009-06-04 | Dsm Ip Assets Bv | Sterol extraction with polar solvent to obtain low sterol and high triglyceride microbial oil |
JP2009179805A (en) * | 1996-03-28 | 2009-08-13 | Dsm Ip Assets Bv | Preparation of microbial polyunsaturated fatty acid-containing oil from biomass sterilized at a low temperature |
-
1994
- 1994-04-20 JP JP6081654A patent/JP2566377B2/en not_active Expired - Fee Related
Cited By (9)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JP2009179805A (en) * | 1996-03-28 | 2009-08-13 | Dsm Ip Assets Bv | Preparation of microbial polyunsaturated fatty acid-containing oil from biomass sterilized at a low temperature |
JP2011024598A (en) * | 1996-03-28 | 2011-02-10 | Dsm Ip Assets Bv | Preparing of microorganism oil including polyunsaturated fatty acid from biomass pasteurized at low temperature |
JP2011132544A (en) * | 1996-03-28 | 2011-07-07 | Dsm Ip Assets Bv | Preparing of microorganism oil including polyunsaturated fatty acid from biomass pasteurized at low temperature |
JP2011130773A (en) * | 1996-03-28 | 2011-07-07 | Dsm Ip Assets Bv | Preparation of microbial polyunsaturated fatty acid containing oil from pasteurised biomass |
JP2011132545A (en) * | 1996-03-28 | 2011-07-07 | Dsm Ip Assets Bv | Preparing of microorganism oil including polyunsaturated fatty acid from biomass pasteurized at low temperature |
JP2009120840A (en) * | 1996-05-15 | 2009-06-04 | Dsm Ip Assets Bv | Sterol extraction with polar solvent to obtain low sterol and high triglyceride microbial oil |
JP2013028808A (en) * | 1996-05-15 | 2013-02-07 | Dsm Ip Assets Bv | Sterol extraction with polar solvent to give low sterol and high triglyceride microbial oil |
JP2014177633A (en) * | 1996-05-15 | 2014-09-25 | Dsm Ip Assets Bv | Sterol extraction with polar solvent to give low sterol, high triglyceride, microbial oil |
WO2006064317A1 (en) * | 2004-12-14 | 2006-06-22 | Avestha Gengraine Technologies Pvt. Ltd. | Recombinant production docosahexaenoic acid (dha) in yeast |
Also Published As
Publication number | Publication date |
---|---|
JP2566377B2 (en) | 1996-12-25 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
CA1321767C (en) | Process for production of eicosapentaenoic acid | |
US5246841A (en) | Microbial process for production of eicosapentaenoic acid | |
JP3425622B2 (en) | Cultures containing polyunsaturated fatty acids and methods for producing oils and fats containing polyunsaturated fatty acids using Labyrinthula | |
US5093249A (en) | Process for production of dihomo-γ-linolenic acid and inhibitor for unsaturation reaction at Δ5-position of fatty acid | |
US20130096332A1 (en) | Method for increasing the content of docosahexaenoic acid in fat-containing materials or in fats and oils | |
CA2577676A1 (en) | Process for production of microbial fat/oil containing discretional amount of diacylglycerol and said fat/oil | |
EP0535939A1 (en) | Process for production of omega 9 type polyunsaturated fatty acid | |
JPS59118090A (en) | Preparation of wax ester, higher fatty alcohol and higher fatty acid | |
JPS6344891A (en) | Production of arachidonic acid | |
AU743992B2 (en) | Process for producing omega-9 highly unsaturated fatty acid and lipid containing the same | |
JP2566377B2 (en) | Method for producing fats and oils high in docosahexaenoic acid | |
CA2121986A1 (en) | Processes for culturing marine microalgae and producing docosahexaenoic acid using the same | |
JP3123789B2 (en) | Method for producing fats and oils and microorganisms therefor | |
JP3071088B2 (en) | Method for producing fats and oils and microorganisms used therefor | |
JPS61177990A (en) | Production of highly unsaturated fatty acid | |
JPH06245759A (en) | Microorganism having oil and fat productivity and production of oil and fat | |
JP2656446B2 (en) | Manufacturing method of squalene | |
JP4045403B2 (en) | Method for producing hydroxy fatty acid and γ-lactone | |
JP4081794B2 (en) | Novel Labyrinthula microorganism and method for producing 4,7,10,13,16-docosapentaenoic acid using the same | |
JPH03272692A (en) | New highly unsaturated fatty acid and production of same fatty acid or lipid containing same fatty acid | |
JPH11243981A (en) | Production of oil and fat containing arachidonic acid and/or eicosapentaenoic acid | |
JPS58165795A (en) | Production of long-chain dicarboxylic acid from fat or oil | |
JP2848810B2 (en) | Method for producing arachidonic acid-rich fat | |
JPH08294384A (en) | Cultivation of seed of marine microalga | |
JPH0787988A (en) | Culture of marine fine algae |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
LAPS | Cancellation because of no payment of annual fees |