JP2020174570A - Method for producing polyunsaturated fatty acid and medium chain fatty acid-containing triglyceride - Google Patents
Method for producing polyunsaturated fatty acid and medium chain fatty acid-containing triglyceride Download PDFInfo
- Publication number
- JP2020174570A JP2020174570A JP2019078911A JP2019078911A JP2020174570A JP 2020174570 A JP2020174570 A JP 2020174570A JP 2019078911 A JP2019078911 A JP 2019078911A JP 2019078911 A JP2019078911 A JP 2019078911A JP 2020174570 A JP2020174570 A JP 2020174570A
- Authority
- JP
- Japan
- Prior art keywords
- fatty acid
- medium
- highly unsaturated
- chain fatty
- 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
- 150000004667 medium chain fatty acids Chemical class 0.000 title claims abstract description 139
- UFTFJSFQGQCHQW-UHFFFAOYSA-N triformin Chemical compound O=COCC(OC=O)COC=O UFTFJSFQGQCHQW-UHFFFAOYSA-N 0.000 title claims abstract description 79
- 238000004519 manufacturing process Methods 0.000 title claims abstract description 34
- 235000020777 polyunsaturated fatty acids Nutrition 0.000 title claims abstract description 19
- 108090001060 Lipase Proteins 0.000 claims abstract description 70
- 239000004367 Lipase Substances 0.000 claims abstract description 70
- 102000004882 Lipase Human genes 0.000 claims abstract description 70
- 235000019421 lipase Nutrition 0.000 claims abstract description 70
- LDVVTQMJQSCDMK-UHFFFAOYSA-N 1,3-dihydroxypropan-2-yl formate Chemical compound OCC(CO)OC=O LDVVTQMJQSCDMK-UHFFFAOYSA-N 0.000 claims abstract description 60
- 235000021122 unsaturated fatty acids Nutrition 0.000 claims description 139
- 150000004670 unsaturated fatty acids Chemical class 0.000 claims description 139
- 238000000034 method Methods 0.000 abstract description 16
- 238000006243 chemical reaction Methods 0.000 description 88
- 239000000243 solution Substances 0.000 description 49
- 239000003921 oil Substances 0.000 description 47
- 235000019198 oils Nutrition 0.000 description 47
- 239000002253 acid Substances 0.000 description 23
- 239000003925 fat Substances 0.000 description 22
- 235000019197 fats Nutrition 0.000 description 22
- 238000004458 analytical method Methods 0.000 description 21
- MBMBGCFOFBJSGT-KUBAVDMBSA-N docosahexaenoic acid Natural products 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 description 19
- 150000004665 fatty acids Chemical class 0.000 description 19
- 108090000790 Enzymes Proteins 0.000 description 18
- 102000004190 Enzymes Human genes 0.000 description 18
- 235000020669 docosahexaenoic acid Nutrition 0.000 description 18
- 235000020673 eicosapentaenoic acid Nutrition 0.000 description 17
- 239000000203 mixture Substances 0.000 description 17
- 235000014113 dietary fatty acids Nutrition 0.000 description 16
- 229930195729 fatty acid Natural products 0.000 description 16
- 239000000194 fatty acid Substances 0.000 description 16
- 238000005886 esterification reaction Methods 0.000 description 13
- 150000002632 lipids Chemical class 0.000 description 13
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 12
- KWYUFKZDYYNOTN-UHFFFAOYSA-M Potassium hydroxide Chemical compound [OH-].[K+] KWYUFKZDYYNOTN-UHFFFAOYSA-M 0.000 description 12
- 150000003626 triacylglycerols Chemical class 0.000 description 12
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 description 10
- OGBUMNBNEWYMNJ-UHFFFAOYSA-N batilol Chemical class CCCCCCCCCCCCCCCCCCOCC(O)CO OGBUMNBNEWYMNJ-UHFFFAOYSA-N 0.000 description 10
- 230000035484 reaction time Effects 0.000 description 10
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 10
- 230000000052 comparative effect Effects 0.000 description 8
- 150000002148 esters Chemical class 0.000 description 8
- WWZKQHOCKIZLMA-UHFFFAOYSA-N octanoic acid Chemical compound CCCCCCCC(O)=O WWZKQHOCKIZLMA-UHFFFAOYSA-N 0.000 description 8
- 238000005809 transesterification reaction Methods 0.000 description 8
- 108010093096 Immobilized Enzymes Proteins 0.000 description 7
- 238000011156 evaluation Methods 0.000 description 7
- 238000005259 measurement Methods 0.000 description 7
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 6
- 241001125048 Sardina Species 0.000 description 6
- GHVNFZFCNZKVNT-UHFFFAOYSA-N decanoic acid Chemical compound CCCCCCCCCC(O)=O GHVNFZFCNZKVNT-UHFFFAOYSA-N 0.000 description 6
- 235000019512 sardine Nutrition 0.000 description 6
- 108010048733 Lipozyme Proteins 0.000 description 5
- 125000004432 carbon atom Chemical group C* 0.000 description 5
- 239000000758 substrate Substances 0.000 description 5
- 239000005635 Caprylic acid (CAS 124-07-2) Substances 0.000 description 4
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 4
- 235000002597 Solanum melongena Nutrition 0.000 description 4
- QTBSBXVTEAMEQO-UHFFFAOYSA-N acetic acid Substances CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 4
- 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 4
- 239000006227 byproduct Substances 0.000 description 4
- 150000001875 compounds Chemical class 0.000 description 4
- 230000032050 esterification Effects 0.000 description 4
- 235000021588 free fatty acids Nutrition 0.000 description 4
- FCCDDURTIIUXBY-UHFFFAOYSA-N lipoamide Chemical compound NC(=O)CCCCC1CCSS1 FCCDDURTIIUXBY-UHFFFAOYSA-N 0.000 description 4
- 125000002496 methyl group Chemical group [H]C([H])([H])* 0.000 description 4
- 239000011259 mixed solution Substances 0.000 description 4
- 229960002446 octanoic acid Drugs 0.000 description 4
- 238000000746 purification Methods 0.000 description 4
- 239000005632 Capric acid (CAS 334-48-5) Substances 0.000 description 3
- 241000222175 Diutina rugosa Species 0.000 description 3
- 241000589516 Pseudomonas Species 0.000 description 3
- 239000000969 carrier Substances 0.000 description 3
- -1 cellite Substances 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- 125000001495 ethyl group Chemical group [H]C([H])([H])C([H])([H])* 0.000 description 3
- 235000013305 food Nutrition 0.000 description 3
- 238000004817 gas chromatography Methods 0.000 description 3
- 238000006460 hydrolysis reaction Methods 0.000 description 3
- 239000000463 material Substances 0.000 description 3
- VLKZOEOYAKHREP-UHFFFAOYSA-N n-Hexane Chemical compound CCCCCC VLKZOEOYAKHREP-UHFFFAOYSA-N 0.000 description 3
- 239000002245 particle Substances 0.000 description 3
- 230000001766 physiological effect Effects 0.000 description 3
- 238000012746 preparative thin layer chromatography Methods 0.000 description 3
- 239000000047 product Substances 0.000 description 3
- 230000002441 reversible effect Effects 0.000 description 3
- 239000002904 solvent Substances 0.000 description 3
- 238000010561 standard procedure Methods 0.000 description 3
- 238000007655 standard test method Methods 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- 238000004809 thin layer chromatography Methods 0.000 description 3
- YUFFSWGQGVEMMI-JLNKQSITSA-N (7Z,10Z,13Z,16Z,19Z)-docosapentaenoic acid Chemical compound CC\C=C/C\C=C/C\C=C/C\C=C/C\C=C/CCCCCC(O)=O YUFFSWGQGVEMMI-JLNKQSITSA-N 0.000 description 2
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 description 2
- 241000228212 Aspergillus Species 0.000 description 2
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- VTYYLEPIZMXCLO-UHFFFAOYSA-L Calcium carbonate Chemical compound [Ca+2].[O-]C([O-])=O VTYYLEPIZMXCLO-UHFFFAOYSA-L 0.000 description 2
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 2
- 241000195493 Cryptophyta Species 0.000 description 2
- 241000235403 Rhizomucor miehei Species 0.000 description 2
- 238000010521 absorption reaction Methods 0.000 description 2
- 235000021342 arachidonic acid Nutrition 0.000 description 2
- 229940114079 arachidonic acid Drugs 0.000 description 2
- 239000008280 blood Substances 0.000 description 2
- 210000004369 blood Anatomy 0.000 description 2
- 239000000919 ceramic Substances 0.000 description 2
- 239000000470 constituent Substances 0.000 description 2
- 230000029087 digestion Effects 0.000 description 2
- 239000003814 drug Substances 0.000 description 2
- 235000021323 fish oil Nutrition 0.000 description 2
- 229940013317 fish oils Drugs 0.000 description 2
- IPCSVZSSVZVIGE-UHFFFAOYSA-N hexadecanoic acid Chemical compound CCCCCCCCCCCCCCCC(O)=O IPCSVZSSVZVIGE-UHFFFAOYSA-N 0.000 description 2
- FUZZWVXGSFPDMH-UHFFFAOYSA-N hexanoic acid Chemical compound CCCCCC(O)=O FUZZWVXGSFPDMH-UHFFFAOYSA-N 0.000 description 2
- 125000002887 hydroxy group Chemical group [H]O* 0.000 description 2
- 239000003456 ion exchange resin Substances 0.000 description 2
- 229920003303 ion-exchange polymer Polymers 0.000 description 2
- 239000012528 membrane Substances 0.000 description 2
- BDAGIHXWWSANSR-UHFFFAOYSA-N methanoic acid Natural products OC=O BDAGIHXWWSANSR-UHFFFAOYSA-N 0.000 description 2
- 238000000199 molecular distillation Methods 0.000 description 2
- 239000002994 raw material Substances 0.000 description 2
- 239000011347 resin Substances 0.000 description 2
- 229920005989 resin Polymers 0.000 description 2
- 238000001179 sorption measurement Methods 0.000 description 2
- 239000006228 supernatant Substances 0.000 description 2
- 238000010998 test method Methods 0.000 description 2
- WRIDQFICGBMAFQ-UHFFFAOYSA-N (E)-8-Octadecenoic acid Natural products CCCCCCCCCC=CCCCCCCC(O)=O WRIDQFICGBMAFQ-UHFFFAOYSA-N 0.000 description 1
- NWUYHJFMYQTDRP-UHFFFAOYSA-N 1,2-bis(ethenyl)benzene;1-ethenyl-2-ethylbenzene;styrene Chemical compound C=CC1=CC=CC=C1.CCC1=CC=CC=C1C=C.C=CC1=CC=CC=C1C=C NWUYHJFMYQTDRP-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
- OSWFIVFLDKOXQC-UHFFFAOYSA-N 4-(3-methoxyphenyl)aniline Chemical compound COC1=CC=CC(C=2C=CC(N)=CC=2)=C1 OSWFIVFLDKOXQC-UHFFFAOYSA-N 0.000 description 1
- QSBYPNXLFMSGKH-UHFFFAOYSA-N 9-Heptadecensaeure Natural products CCCCCCCC=CCCCCCCCC(O)=O QSBYPNXLFMSGKH-UHFFFAOYSA-N 0.000 description 1
- 241000590020 Achromobacter Species 0.000 description 1
- 241000932047 Achromobacter sp. Species 0.000 description 1
- 241000588810 Alcaligenes sp. Species 0.000 description 1
- 240000006439 Aspergillus oryzae Species 0.000 description 1
- 235000002247 Aspergillus oryzae Nutrition 0.000 description 1
- 241000894006 Bacteria Species 0.000 description 1
- 241001474374 Blennius Species 0.000 description 1
- 241000273930 Brevoortia tyrannus Species 0.000 description 1
- 241001508395 Burkholderia sp. Species 0.000 description 1
- 241000222120 Candida <Saccharomycetales> Species 0.000 description 1
- 241000238366 Cephalopoda Species 0.000 description 1
- 241000199913 Crypthecodinium Species 0.000 description 1
- 235000021294 Docosapentaenoic acid Nutrition 0.000 description 1
- 241000195619 Euglena gracilis Species 0.000 description 1
- 241000233866 Fungi Species 0.000 description 1
- OPGOLNDOMSBSCW-CLNHMMGSSA-N Fursultiamine hydrochloride Chemical compound Cl.C1CCOC1CSSC(\CCO)=C(/C)N(C=O)CC1=CN=C(C)N=C1N OPGOLNDOMSBSCW-CLNHMMGSSA-N 0.000 description 1
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-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
- 241001465754 Metazoa Species 0.000 description 1
- 241001661345 Moesziomyces antarcticus Species 0.000 description 1
- 241000907999 Mortierella alpina Species 0.000 description 1
- 241000498617 Mucor javanicus Species 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
- 235000021314 Palmitic acid Nutrition 0.000 description 1
- ISWSIDIOOBJBQZ-UHFFFAOYSA-N Phenol Chemical compound OC1=CC=CC=C1 ISWSIDIOOBJBQZ-UHFFFAOYSA-N 0.000 description 1
- 241000589774 Pseudomonas sp. Species 0.000 description 1
- 240000005384 Rhizopus oryzae Species 0.000 description 1
- 235000013752 Rhizopus oryzae Nutrition 0.000 description 1
- 241000269851 Sarda sarda Species 0.000 description 1
- 235000021355 Stearic acid Nutrition 0.000 description 1
- 241000282898 Sus scrofa Species 0.000 description 1
- 241000223258 Thermomyces lanuginosus Species 0.000 description 1
- 241001261506 Undaria pinnatifida Species 0.000 description 1
- JAZBEHYOTPTENJ-JLNKQSITSA-N all-cis-5,8,11,14,17-icosapentaenoic acid Chemical compound CC\C=C/C\C=C/C\C=C/C\C=C/C\C=C/CCCC(O)=O JAZBEHYOTPTENJ-JLNKQSITSA-N 0.000 description 1
- DTOSIQBPPRVQHS-PDBXOOCHSA-N alpha-linolenic acid Chemical compound CC\C=C/C\C=C/C\C=C/CCCCCCCC(O)=O DTOSIQBPPRVQHS-PDBXOOCHSA-N 0.000 description 1
- 235000020661 alpha-linolenic acid Nutrition 0.000 description 1
- 239000010775 animal oil Substances 0.000 description 1
- 239000011324 bead Substances 0.000 description 1
- 229910000019 calcium carbonate Inorganic materials 0.000 description 1
- 238000004364 calculation method Methods 0.000 description 1
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 description 1
- 150000001732 carboxylic acid derivatives Chemical class 0.000 description 1
- 239000012159 carrier gas Substances 0.000 description 1
- 239000003054 catalyst Substances 0.000 description 1
- 239000003026 cod liver oil Substances 0.000 description 1
- 235000012716 cod liver oil Nutrition 0.000 description 1
- 230000003920 cognitive function Effects 0.000 description 1
- 239000002131 composite material Substances 0.000 description 1
- 239000002537 cosmetic Substances 0.000 description 1
- 238000004132 cross linking Methods 0.000 description 1
- 238000000354 decomposition reaction Methods 0.000 description 1
- 230000006866 deterioration Effects 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 238000002845 discoloration Methods 0.000 description 1
- 229940090949 docosahexaenoic acid Drugs 0.000 description 1
- 229960005135 eicosapentaenoic acid Drugs 0.000 description 1
- JAZBEHYOTPTENJ-UHFFFAOYSA-N eicosapentaenoic acid Natural products CCC=CCC=CCC=CCC=CCC=CCCCC(O)=O JAZBEHYOTPTENJ-UHFFFAOYSA-N 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 238000001704 evaporation Methods 0.000 description 1
- 235000019253 formic acid Nutrition 0.000 description 1
- 239000011521 glass Substances 0.000 description 1
- 125000005456 glyceride group Chemical group 0.000 description 1
- 239000001307 helium Substances 0.000 description 1
- 229910052734 helium Inorganic materials 0.000 description 1
- SWQJXJOGLNCZEY-UHFFFAOYSA-N helium atom Chemical compound [He] SWQJXJOGLNCZEY-UHFFFAOYSA-N 0.000 description 1
- 239000001257 hydrogen Substances 0.000 description 1
- 229910052739 hydrogen Inorganic materials 0.000 description 1
- 230000007062 hydrolysis Effects 0.000 description 1
- 230000002209 hydrophobic effect Effects 0.000 description 1
- 230000002779 inactivation Effects 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 150000002500 ions 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
- 229960004488 linolenic acid Drugs 0.000 description 1
- KQQKGWQCNNTQJW-UHFFFAOYSA-N linolenic acid Natural products CC=CCCC=CCC=CCCCCCCCC(O)=O KQQKGWQCNNTQJW-UHFFFAOYSA-N 0.000 description 1
- 235000021056 liquid food Nutrition 0.000 description 1
- 238000012423 maintenance Methods 0.000 description 1
- 238000000691 measurement method Methods 0.000 description 1
- JBXYCUKPDAAYAS-UHFFFAOYSA-N methanol;trifluoroborane Chemical compound OC.FB(F)F JBXYCUKPDAAYAS-UHFFFAOYSA-N 0.000 description 1
- 230000000813 microbial effect Effects 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 239000012046 mixed solvent Substances 0.000 description 1
- WQEPLUUGTLDZJY-UHFFFAOYSA-N n-Pentadecanoic acid Natural products CCCCCCCCCCCCCCC(O)=O WQEPLUUGTLDZJY-UHFFFAOYSA-N 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- QIQXTHQIDYTFRH-UHFFFAOYSA-N octadecanoic acid Chemical compound CCCCCCCCCCCCCCCCCC(O)=O QIQXTHQIDYTFRH-UHFFFAOYSA-N 0.000 description 1
- OQCDKBAXFALNLD-UHFFFAOYSA-N octadecanoic acid Natural products CCCCCCCC(C)CCCCCCCCC(O)=O OQCDKBAXFALNLD-UHFFFAOYSA-N 0.000 description 1
- ZQPPMHVWECSIRJ-KTKRTIGZSA-N oleic acid Chemical compound CCCCCCCC\C=C/CCCCCCCC(O)=O ZQPPMHVWECSIRJ-KTKRTIGZSA-N 0.000 description 1
- 235000021313 oleic acid Nutrition 0.000 description 1
- 235000008390 olive oil Nutrition 0.000 description 1
- 239000004006 olive oil Substances 0.000 description 1
- 210000000496 pancreas Anatomy 0.000 description 1
- KJFMBFZCATUALV-UHFFFAOYSA-N phenolphthalein Chemical compound C1=CC(O)=CC=C1C1(C=2C=CC(O)=CC=2)C2=CC=CC=C2C(=O)O1 KJFMBFZCATUALV-UHFFFAOYSA-N 0.000 description 1
- 230000000704 physical effect Effects 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 229940119224 salmon oil Drugs 0.000 description 1
- 238000005070 sampling Methods 0.000 description 1
- 235000003441 saturated fatty acids Nutrition 0.000 description 1
- 150000004671 saturated fatty acids Chemical class 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 239000008117 stearic acid Substances 0.000 description 1
- JIWBIWFOSCKQMA-UHFFFAOYSA-N stearidonic acid Natural products CCC=CCC=CCC=CCC=CCCCCC(O)=O JIWBIWFOSCKQMA-UHFFFAOYSA-N 0.000 description 1
- 239000013589 supplement Substances 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 238000004448 titration Methods 0.000 description 1
- 125000005457 triglyceride group Chemical group 0.000 description 1
- 235000015112 vegetable and seed oil Nutrition 0.000 description 1
- 239000008158 vegetable oil Substances 0.000 description 1
Abstract
Description
本発明は、高度不飽和脂肪酸を含有する、モノグリセリド及びジグリセリドを、中鎖脂肪酸存在下でリパーゼ処理する高度不飽和脂肪酸及び中鎖脂肪酸含有トリグリセリドの製造方法であって、前記モノグリセリド及びジグリセリドの質量比が２０：８０〜８５：１５であることを特徴とする、高度不飽和脂肪酸及び中鎖脂肪酸含有トリグリセリドの製造方法に関する。 The present invention is a method for producing a highly unsaturated fatty acid and a medium-chain fatty acid-containing triglyceride in which a monoglyceride and a diglyceride containing a highly unsaturated fatty acid are treated with lipase in the presence of a medium-chain fatty acid, and the mass ratio of the monoglyceride and the diglyceride. The present invention relates to a method for producing a highly unsaturated fatty acid and a medium chain fatty acid-containing triglyceride, which comprises 20:80 to 85:15.
高度不飽和脂肪酸及びその誘導体は、血中脂肪の低減等の多くの生理活性を有し、古くから医薬品、化粧品、食品等の原料として使用されており、その中には、種々の生理活性を有するものが知られており、国内外において医薬品や特定保健用食品、機能性表示食品、サプリメント等としての需要が高い。
しかしながら、高度不飽和脂肪酸を構成脂肪酸とする天然の油脂は、通常パルミチン酸やステアリン酸、オレイン酸といったその他の脂肪酸も含むトリグリセリドを構成しており、天然の油脂を摂取することで上記のような効果を得るためには多量の摂取量が必要となる。
Highly unsaturated fatty acids and their derivatives have many physiological activities such as reduction of blood fat, and have been used as raw materials for pharmaceuticals, cosmetics, foods, etc. for a long time, and various physiological activities are contained in them. It is known to have, and is in high demand in Japan and overseas as pharmaceuticals, foods for specified health uses, foods with functional claims, supplements, etc.
However, natural fats and oils containing highly unsaturated fatty acids as constituent fatty acids usually constitute triglycerides including other fatty acids such as palmitic acid, stearic acid, and oleic acid, and by ingesting natural fats and oils, as described above. A large amount of intake is required to obtain the effect.
そこで、高度不飽和脂肪酸を構成脂肪酸とする天然の油脂にエチルエステル化処理を施し、脂肪酸種の物理的特性の違いを利用して高度不飽和脂肪酸のエチルエステル化物を高純度化する方法が開発されている。しかしながら、トリグリセリドのようなアシルグリセロールと比べてエチルエステル化物は体内への吸収性が悪いことが知られている。
そのため、体内への吸収性の優れた高度不飽和脂肪酸含有トリグリセリドの製造方法が求められている。
Therefore, a method has been developed in which natural fats and oils containing highly unsaturated fatty acids as constituent fatty acids are subjected to ethyl esterification treatment, and the ethyl esterified products of highly unsaturated fatty acids are purified by utilizing the difference in physical properties of fatty acid species. Has been done. However, it is known that ethyl esterified products are less absorbable into the body than acylglycerols such as triglycerides.
Therefore, there is a demand for a method for producing a highly unsaturated fatty acid-containing triglyceride having excellent absorbability into the body.
この課題の解決策の1つに、中鎖脂肪酸の利用がある。
中鎖脂肪酸は消化吸収に優れていることが古くから知られており、流動食の脂質源などとして利用されている。
このため、中鎖脂肪酸を組み込んだ高度不飽和脂肪酸含有トリグリセリド含有組成物を得ることを目的とした製造方法の試みが行われている。
例えば、中鎖脂肪酸を高度不飽和脂肪酸含有トリグリセリドと混合した原料油脂に対し、リパーゼを作用させてエステル交換反応を行う製造方法が提案されている（特許文献１：特開平８−２１４８９１）。
One of the solutions to this problem is the use of medium-chain fatty acids.
Medium-chain fatty acids have long been known to be excellent in digestion and absorption, and are used as a lipid source for liquid foods.
For this reason, attempts have been made on a production method for the purpose of obtaining a polyunsaturated fatty acid-containing triglyceride-containing composition incorporating a medium-chain fatty acid.
For example, a production method has been proposed in which a transesterification reaction is carried out by allowing lipase to act on a raw material fat or oil in which a medium-chain fatty acid is mixed with a triglyceride containing a highly unsaturated fatty acid (Patent Document 1: Japanese Patent Application Laid-Open No. 8-214891).
しかしながら、エステル交換反応が可逆的な反応のため、エステル交換反応を効率良く進めるために大量の基質（特許文献１では遊離中鎖脂肪酸）を使用する必要があり、例えば生産設備に大型のものが必要になる、反応副産物や未反応の遊離脂肪酸の量が多く精製や廃棄にコストがかかる、などといった産業的な課題がある。 However, since the transesterification reaction is a reversible reaction, it is necessary to use a large amount of substrate (free medium-chain fatty acid in Patent Document 1) in order to efficiently proceed with the transesterification reaction. There are industrial issues such as the need for a large amount of reaction by-products and unreacted free fatty acids, which makes purification and disposal costly.
また、高度不飽和脂肪酸含有モノグリセリドにリパーゼを作用させて中鎖脂肪酸を導入するエステル化反応を利用する方法が提案されているが（特許文献２：特開２００２−２７９９５号）、エステル化反応の可逆反応である加水分解反応が起きるために反応が進みづらいという課題がある。
これを解決するためには、エステル化反応の副産物である水を高温減圧条件で蒸発させながら反応を進めることが考えられる。
しかし高温条件下ではリパーゼが不活化してしまう、高度不飽和脂肪酸に熱履歴が生じて品質が損なわれる、といった別の課題が生じる。
そのため、比較的低温な条件でエステル化反応を進める場合には、特許文献２のように、エステル交換反応と同様に大量の基質が必要となる。
Further, a method using an esterification reaction in which a lipase is allowed to act on a highly unsaturated fatty acid-containing monoglyceride to introduce a medium-chain fatty acid has been proposed (Patent Document 2: Japanese Patent Application Laid-Open No. 2002-27995), but the esterification reaction There is a problem that the reaction is difficult to proceed because the hydrolysis reaction, which is a reversible reaction, occurs.
In order to solve this, it is conceivable to proceed with the reaction while evaporating water, which is a by-product of the esterification reaction, under high temperature and reduced pressure conditions.
However, there are other problems such as inactivation of lipase under high temperature conditions and deterioration of quality due to thermal history of polyunsaturated fatty acids.
Therefore, when the esterification reaction is carried out under relatively low temperature conditions, a large amount of substrate is required as in the transesterification reaction as in Patent Document 2.
そこで、本発明の目的は、高度不飽和脂肪酸を含有する、モノグリセリド及びジグリセリドから効率的に高度不飽和脂肪酸及び中鎖脂肪酸含有トリグリセリドを製造できる方法を提供するものである。 Therefore, an object of the present invention is to provide a method capable of efficiently producing highly unsaturated fatty acid and medium chain fatty acid-containing triglyceride from monoglyceride and diglyceride containing highly unsaturated fatty acid.
すなわち、本発明は、
（１）高度不飽和脂肪酸を含有する、モノグリセリド及びジグリセリドを、中鎖脂肪酸存在下でリパーゼ処理する高度不飽和脂肪酸及び中鎖脂肪酸含有トリグリセリドの製造方法であって、
前記モノグリセリド及びジグリセリドの質量比が２０：８０〜８５：１５であることを特徴とする、
高度不飽和脂肪酸及び中鎖脂肪酸含有トリグリセリドの製造方法、
（２）前記中鎖脂肪酸が前記モノグリセリド及びジグリセリドの合計１００質量部に対し１０質量部以上１００質量部以下であることを特徴とする、
（１）記載の高度不飽和脂肪酸及び中鎖脂肪酸含有トリグリセリドの製造方法、
である。
That is, the present invention
(1) A method for producing a highly unsaturated fatty acid and a medium-chain fatty acid-containing triglyceride, in which a monoglyceride and a diglyceride containing a highly unsaturated fatty acid are lipase-treated in the presence of a medium-chain fatty acid.
The mass ratio of the monoglyceride to the diglyceride is 20:80 to 85:15.
Method for producing highly unsaturated fatty acid and medium chain fatty acid-containing triglyceride,
(2) The medium-chain fatty acid is 10 parts by mass or more and 100 parts by mass or less with respect to 100 parts by mass in total of the monoglyceride and diglyceride.
(1) Method for producing highly unsaturated fatty acid and medium chain fatty acid-containing triglyceride,
Is.
本発明は、高度不飽和脂肪酸を含有する、モノグリセリド及びジグリセリドから効率的に高度不飽和脂肪酸及び中鎖脂肪酸含有トリグリセリドを製造できる方法を提供するものである。 The present invention provides a method capable of efficiently producing highly unsaturated fatty acid and medium chain fatty acid-containing triglyceride from monoglyceride and diglyceride containing highly unsaturated fatty acid.
以下、本発明を詳細に説明する。
なお、本発明において、「％」は「質量％」を、「部」は「質量部」を意味する。
Hereinafter, the present invention will be described in detail.
In the present invention, "%" means "% by mass" and "part" means "part by mass".
＜本発明の特徴＞
本発明の製造方法は、高度不飽和脂肪酸を含有する、モノグリセリド及びジグリセリドを、中鎖脂肪酸存在下でリパーゼ処理する高度不飽和脂肪酸及び中鎖脂肪酸含有トリグリセリドの製造方法であって、
前記モノグリセリド及びジグリセリドの質量比が２０：８０〜８５：１５である、高度不飽和脂肪酸及び中鎖脂肪酸含有トリグリセリドの製造方法に特徴を有する。
<Features of the present invention>
The production method of the present invention is a method for producing a highly unsaturated fatty acid and a medium chain fatty acid-containing triglyceride in which monoglycerides and diglycerides containing highly unsaturated fatty acids are lipase-treated in the presence of medium-chain fatty acids.
It is characterized by a method for producing a highly unsaturated fatty acid and a medium-chain fatty acid-containing triglyceride having a mass ratio of monoglyceride and diglyceride of 20:80 to 85:15.
＜高度不飽和脂肪酸を含有する、モノグリセリド及びジグリセリド＞
本発明の製造方法において、高度不飽和脂肪酸を含有する、モノグリセリド及びジグリセリドとは、モノグリセリドを構成する脂肪酸残基が高度不飽和脂肪酸である化合物及び、ジグリセリドを構成する脂肪酸残基の一部又は全部が高度不飽和脂肪酸である化合物をいうが、高度不飽和脂肪酸を含有する、モノグリセリド及びジグリセリド以外のその他の成分を含んでもよい。
また高度不飽和脂肪酸を含有する、モノグリセリド及びジグリセリドに加え、高度不飽和脂肪酸を含有しない、モノグリセリド及びジグリセリド等を含んでも良い。
なお、高度不飽和脂肪酸を含有する、モノグリセリド及びジグリセリドに含まれるDHAの含有量としては、高度不飽和脂肪酸を含有する、モノグリセリド及びジグリセリド中に１０％以上４０％以下であればよい。
また、高度不飽和脂肪酸を含有する、モノグリセリド及びジグリセリドに含まれるEPAの含有量としては、高度不飽和脂肪酸を含有する、モノグリセリド及びジグリセリド中に５％以上３５％以下であればよい。
なお、効率的に高度不飽和脂肪酸及び中鎖脂肪酸含有トリグリセリドを製造しにくくなるため、その他の成分の割合は、高度不飽和脂肪酸を含有する、モノグリセリド及びジグリセリドの合計１００質量部に対し５０質量部以下であるとよく、さらに３０質量部以下であるとよく、特に２０質量部以下であるとよい。本発明において、高度不飽和脂肪酸とは、炭素数が１８以上、かつ分子内に二重結合を２個以上有した不飽和脂肪酸を意味し、例えば、ドコサヘキサエン酸（C22:6、DHA）、エイコサペンタエン酸（C20:5、EPA）、アラキドン酸（C20:4、AA）、ドコサペンタエン酸（C22:5、DPA）、ステアリドン酸（C18:4）、リノレン酸（C18:3）、リノール酸（C18:2）等が挙げられる。
また、高度不飽和脂肪酸を含有する、モノグリセリド及びジグリセリドに含まれる高度不飽和脂肪酸としては、血中脂肪の低減等の多くの生理活性を有することからDHA又はEPAがよく、さらに認知機能の維持等の作用もあることからDHAが良い。
なお、高度不飽和脂肪酸及び中鎖脂肪酸含有トリグリセリドに含まれるDHAの含有量としては、高度不飽和脂肪酸及び中鎖脂肪酸含有トリグリセリド中に１０％以上４０％以下であればよい。
また、高度不飽和脂肪酸及び中鎖脂肪酸含有トリグリセリドに含まれるEPAの含有量としては、高度不飽和脂肪酸及び中鎖脂肪酸含有トリグリセリド中に５％以上３５％以下であればよい。
<Monoglycerides and diglycerides containing highly unsaturated fatty acids>
In the production method of the present invention, monoglycerides and diglycerides containing highly unsaturated fatty acids are compounds in which the fatty acid residues constituting the monoglycerides are highly unsaturated fatty acids, and some or all of the fatty acid residues constituting the diglycerides. Is a highly unsaturated fatty acid, but may contain other components other than monoglycerides and diglycerides, which contain highly unsaturated fatty acids.
Further, in addition to monoglyceride and diglyceride containing highly unsaturated fatty acid, monoglyceride and diglyceride not containing highly unsaturated fatty acid may be contained.
The content of DHA contained in the monoglyceride and diglyceride containing highly unsaturated fatty acid may be 10% or more and 40% or less in the monoglyceride and diglyceride containing highly unsaturated fatty acid.
The content of EPA contained in the monoglyceride and diglyceride containing highly unsaturated fatty acid may be 5% or more and 35% or less in the monoglyceride and diglyceride containing highly unsaturated fatty acid.
Since it becomes difficult to efficiently produce highly unsaturated fatty acid and medium chain fatty acid-containing triglyceride, the ratio of other components is 50 parts by mass with respect to 100 parts by mass of the total of monoglyceride and diglyceride containing highly unsaturated fatty acid. It is preferably less than or equal to, more preferably 30 parts by mass or less, and particularly preferably 20 parts by mass or less. In the present invention, the highly unsaturated fatty acid means an unsaturated fatty acid having 18 or more carbon atoms and having two or more double bonds in the molecule, for example, docosahexaenoic acid (C22: 6, DHA), d. Eicosapentaenoic acid (C20: 5, EPA), arachidonic acid (C20: 4, AA), docosapentaenoic acid (C22: 5, DPA), stearidonic acid (C18: 4), linolenic acid (C18: 3), linoleic acid (C18: 2) and the like.
Further, as the highly unsaturated fatty acid contained in monoglyceride and diglyceride containing highly unsaturated fatty acid, DHA or EPA is preferable because it has many physiological activities such as reduction of blood fat, and further maintenance of cognitive function, etc. DHA is good because it also has the effect of.
The content of DHA contained in the highly unsaturated fatty acid and the medium-chain fatty acid-containing triglyceride may be 10% or more and 40% or less in the highly unsaturated fatty acid and the medium-chain fatty acid-containing triglyceride.
The content of EPA contained in the highly unsaturated fatty acid and the medium-chain fatty acid-containing triglyceride may be 5% or more and 35% or less in the highly unsaturated fatty acid and the medium-chain fatty acid-containing triglyceride.
＜高度不飽和脂肪酸を含有する、モノグリセリド及びジグリセリドの種類＞
高度不飽和脂肪酸を含有する、モノグリセリド及びジグリセリドとしては、例えば、魚油、魚油以外の動物油、植物油、藻類、微生物が生産する油、又はこれらの混合油脂等の由来の分解物が挙げられるが、高度不飽和脂肪酸を含有する、モノグリセリド及びジグリセリドが多く取れることから、魚油がよく、中でもDHAを豊富に含むことからイワシ油又はメンヘーデン油がよい。
<Types of monoglycerides and diglycerides containing highly unsaturated fatty acids>
Examples of monoglycerides and diglycerides containing highly unsaturated fatty acids include fish oils, animal oils other than fish oils, vegetable oils, algae, oils produced by microorganisms, and decomposition products derived from mixed fats and oils thereof. Fish oil is preferable because it contains a large amount of monoglyceride and diglyceride containing unsaturated fatty acids, and sardine oil or menheden oil is particularly preferable because it is rich in DHA.
魚油としては、イワシ油、マグロ油、カツオ油、タラ肝油、サケ油、イカ油、又はメンヘーデン油等が挙げられる。
藻類、微生物由来油としては、Ｍｏｒｔｉｅｒｅｌｌａ ａｌｐｉｎｅ、Ｅｕｇｌｅｎａ ｇｒａｃｉｌｉｓ等によるアラキドン酸含有油、クロメ、アラメ、ワカメ、ヒジキ、ハバノリ、ヒバマタの一種によるＥＰＡ含有油、Ｃｒｙｐｔｈｅｃｏｄｉｎｉｕｍ ｃｏｈｎｉｉ、Ｖｉｂｒｉｏ ｍａｒｉｎｕｓ、Ｔｈｒａｕｓｔｏｃｈｙｔｒｉｕｍ ａｕｒｅｕｍ、Ｓｈｅｗａｎｅｌｌａ属細菌等によるＤＨＡ含有油等が挙げられる。
Examples of fish oil include sardine oil, tuna oil, bonito oil, cod liver oil, salmon oil, squid oil, menhaden oil and the like.
Examples of algae and microbial-derived oils include arachidonic acid-containing oils from Mortierella alpine, Euglena gracilis, etc., EPA-containing oils from kurome, arame, wakame seaweed, hijiki, habanori, and hibamata, Crypthecodinium cohni, Vivi DHA-containing oil and the like due to the above.
＜高度不飽和脂肪酸を含有する、モノグリセリド及びジグリセリドの比率＞
本発明の製造方法に用いる高度不飽和脂肪酸を含有する、モノグリセリド及びジグリセリドの比率は、効率的に高度不飽和脂肪酸及び中鎖脂肪酸含有トリグリセリドを製造できることから、前記モノグリセリド及びジグリセリドの質量比が２０：８０〜８５：１５であるとよい。
<Ratio of monoglyceride and diglyceride containing highly unsaturated fatty acids>
The ratio of monoglyceride to diglyceride containing highly unsaturated fatty acid used in the production method of the present invention is such that the mass ratio of monoglyceride to diglyceride is 20: because highly unsaturated fatty acid and medium-chain fatty acid-containing triglyceride can be efficiently produced. It is preferably 80 to 85:15.
＜高度不飽和脂肪酸及び中鎖脂肪酸含有トリグリセリド＞
本発明において、高度不飽和脂肪酸及び中鎖脂肪酸含有トリグリセリドとは、トリグリセリドを構成する脂肪酸残基の一部又は全部が高度不飽和脂肪酸及び中鎖脂肪酸である化合物をいう。
<Triglycerides containing polyunsaturated fatty acids and medium-chain fatty acids>
In the present invention, the highly unsaturated fatty acid and the medium chain fatty acid-containing triglyceride refer to a compound in which a part or all of the fatty acid residues constituting the triglyceride are the highly unsaturated fatty acid and the medium chain fatty acid.
＜リパーゼ＞
本発明の製造方法に用いるリパーゼは、１，３位−特異的であっても、非特異的であってもよい。
ここで、１，３位特異リパーゼとしては、トリグリセリドの１，３位にのみ特異的に作用する酵素、又は２位よりも１，３位に優先的に作用する酵素である。１，３位に中鎖脂肪酸、２位に高度不飽和脂肪酸が結合したトリグリセリドが吸収性に優れていることから、１，３位特異リパーゼを用いて、高度不飽和脂肪酸を含有する、モノグリセリド及びジグリセリドと、中鎖脂肪酸をエステル結合させると良い。
<Lipase>
The lipase used in the production method of the present invention may be 1,3-position-specific or non-specific.
Here, the 1,3-position specific lipase is an enzyme that specifically acts only on the 1,3-position of triglyceride, or an enzyme that preferentially acts on the 1,3-position over the 2-position. Since triglycerides in which medium-chain fatty acids are bound to the 1st and 3rd positions and highly unsaturated fatty acids are bound to the 2nd position are excellent in absorbability, monoglycerides and monoglycerides containing highly unsaturated fatty acids are used by using the 1,3 position specific lipase. It is preferable to ester-bond the diglyceride and the medium-chain fatty acid.
リパーゼの性状は、粗精製、部分精製、精製のいずれでもよい。また、遊離型でも固定化酵素でもよいが、再利用可能である点、反応後の処置が簡便である点で、固定化酵素を用いるとよい。 The properties of lipase may be crude purification, partial purification, or purification. Further, although it may be a free form or an immobilized enzyme, it is preferable to use an immobilized enzyme because it can be reused and the treatment after the reaction is simple.
＜担体＞
固定化酵素は、酵素を担体、フィルム、又は膜に固定化されたものであってもよい。担体としては、イオン交換樹脂、多孔性樹脂、セラミックス、炭酸カルシウム、セライト、ガラスビーズ、活性炭等の有機担体、無機担体、有機無機複合担体が挙げられる。耐久性やリパーゼとの親和性に優れていることから、担体は、イオン交換樹脂、多孔性樹脂、セラミックスを用いるとよい。
<Carrier>
The immobilized enzyme may be an enzyme immobilized on a carrier, a film, or a membrane. Examples of the carrier include organic carriers such as ion exchange resins, porous resins, ceramics, calcium carbonate, cellite, glass beads, and activated carbon, inorganic carriers, and organic-inorganic composite carriers. Since it is excellent in durability and affinity with lipase, it is preferable to use an ion exchange resin, a porous resin, or ceramics as the carrier.
固定化の方法としては、包括法、架橋法、物理的吸着法、イオン吸着法、共有結合法、疎水結合法等が挙げられる。 Examples of the immobilization method include a comprehensive method, a cross-linking method, a physical adsorption method, an ion adsorption method, a covalent bond method, and a hydrophobic bond method.
＜粒子径＞
固定化酵素が粒子状である場合、粒子状の担体に酵素を固定化することができる。この場合、担体の粒子径が０．０１ｍｍ以上３ｍｍ以下であればよく、さらに０．０５ｍｍ以上１．５ｍｍ以下であればよい。
<Particle diameter>
When the immobilized enzyme is in the form of particles, the enzyme can be immobilized on the particulate carrier. In this case, the particle size of the carrier may be 0.01 mm or more and 3 mm or less, and further 0.05 mm or more and 1.5 mm or less.
＜リパーゼの具体例＞
本発明において、リパーゼとしては、例えば、リゾムコール属（Ｒｈｉｚｏｍｕｃｏｒ ｍｉｅｈｅｉ）、ムコール属(Ｍｕｃｏｒ ｍｉｅｈｅｉ，Ｍｕｃｏｒ ｊａｖａｎｉｃｕｓ)、アスペルギルス属(Ａｓｐｅｒｇｉｌｌｕｓ ｏｒｙｚａｅ，Ａｓｐｅｒｇｉｌｌｕｓ ｎｉｇｅｒ)、リゾプス属(Ｒｈｉｚｏｐｕｓ ｓｐ.)、ペニシリウム属(Ｐｅｎｉｃｉｌｌｉｕｍ ｒｏｑｕｅｆｏｒｔｉ，Ｐｅｎｉｃｉｌｌｉｕｍ ｃａｍｅｍｂｅｒｔｉ)、サーモマイセス属(Ｔｈｅｒｍｏｍｙｃｅｓ ｌａｎｕｇｉｎｏｓｕｓ)等に属する糸状菌、キャンディダ属（Ｃａｎｄｉｄａ ａｎｔａｒｃｔｉｃａ，Ｃａｎｄｉｄａ ｒｕｇｏｓａ，Ｃａｎｄｉｄａ ｃｙｌｉｎｄｒａｃｅａ）、ピヒア（Ｐｉｃｈｉａ）等に属する酵母、シュードモナス属(Ｐｓｅｕｄｏｍｏｎａｓ ｓｐ.)、アクロモバクター属(Ａｃｈｒｏｍｏｂａｃｔｅｒ ｓｐ.)、ブルクホルデリア属(Ｂｕｒｋｈｏｌｄｅｒｉａ ｓｐ．)、アルカリゲネス属（Ａｌｃａｌｉｇｅｎｅｓ ｓｐ．）、シュードザイマ属(Ｐｓｅｕｄｏｚｙｍａ ｓｐ．)等に属する細菌、豚膵臓等の動物に由来するリパーゼが挙げられる。例えば、Ｒｈｉｚｏｐｕｓ ｏｒｙｚａｅのリパーゼ（リパーゼＤＦ：天野エンザイム社製）、Ｃａｎｄｉｄａ ｒｕｇｏｓａ（リパーゼＯＦ：名糖産業社製）およびＰｓｅｕｄｏｍａｎａｓ属のリパーゼ（リパーゼＰＳ、リパーゼＡＫ：天野製薬社製）が挙げられ、リパーゼの固定化酵素としては、Ｒｈｉｚｏｍｕｃｏｒ ｍｉｅｈｅｉのリパーゼ（リポザイムＩＭ６０：ノボザイムズ社製、リポザイムＲＭＩＭ：ノボザイムズ社製、Ｎｏｖｏｚｙｍ （登録商標）４００８６：ノボザイムズ社製）、Ｐｓｅｕｄｏｚｙｍａ ａｎｔａｒｃｔｉｃａのリパーゼ（ノボザイム４３５：ノボザイムズ社製）が挙げられる。
<Specific example of lipase>
In the present invention, as the lipase, for example, Pseudomonas genus (Rhizomucor miehei), Achromobacter genus (Mucor miehei, Mucor javanicus), Aspergillus genus (Aspergillus oryzae, Aspergillus lyspirus) , Pencillium camemberti), filamentous fungi belonging to the genus Thermomyces lanuginosus, etc., genus Candida (Candida antarctica, Candida rugosa, Candida rugosa, genus Pseudomonas, belonging to the genus Pseudomonas Examples include bacteria belonging to the genus Achromobacter sp., The genus Burkholderia sp., The genus Alcaligenes sp., The genus Pseudomonas sp., And lipases derived from animals such as pig pancreas. .. For example, lipase of Rhizopus oryzae (lipase DF: manufactured by Amano Enzyme), Candida rugosa (lipase OF: manufactured by Meizu Sangyo Co., Ltd.) and lipase of the genus Pseudomanas (lipase PS, lipase AK: manufactured by Amano Pharmaceutical Co., Ltd.) can be mentioned. Lipase (Lipozyme IM60: manufactured by Novozymes, Lipozyme RMIM: manufactured by Novozymes, Novozyme (registered trademark) 40086: manufactured by Novozymes), Pseudozymase (Novozymes), Lipase (Made by Novozymes), Lipase (Made by Novozymes) Can be mentioned.
＜リパーゼの使用量＞
リパーゼの使用量は、反応条件によって適宜決定すればよく、特に制限されるものではない。例えば、遊離型の酵素を用いる場合は、一般的には反応液１ｇ当たり１単位(Ｕ)以上１０，０００Ｕ以下添加することができ、さらに５Ｕ以上１，０００Ｕ以下添加することができる。
<Amount of lipase used>
The amount of lipase used may be appropriately determined depending on the reaction conditions and is not particularly limited. For example, when a free enzyme is used, generally, 1 unit (U) or more and 10,000 U or less can be added per 1 g of the reaction solution, and 5 U or more and 1,000 U or less can be added.
ここで、酵素活性の１Ｕとは、リパーゼの場合はオリーブ油の加水分解において１分間に１μｍｏｌの脂肪酸を遊離する酵素量を指す。 Here, 1U of enzyme activity refers to the amount of enzyme that liberates 1 μmol of fatty acid per minute in the hydrolysis of olive oil in the case of lipase.
また、リパーゼの固定化酵素を用いる場合は、上記Ｕにあわせ適宜その使用量を調整すればよいが、工場等において都度のＵによるリパーゼ使用量の計算等の工数の増加を避けるため反応液の質量に対して、担体の質量を含む質量として０．１質量％以上２００質量％以下になるように、さらに１質量％以上３５質量％以下、特に１質量％以上２０質量％以下になるように添加することができる。 When a lipase-immobilized enzyme is used, the amount used may be adjusted as appropriate according to the above U, but in order to avoid an increase in the number of steps such as calculation of the amount of lipase used by U each time in a factory or the like, the reaction solution is used. With respect to the mass, the mass including the mass of the carrier should be 0.1% by mass or more and 200% by mass or less, and further 1% by mass or more and 35% by mass or less, particularly 1% by mass or more and 20% by mass or less. Can be added.
＜酸価＞
本発明において、酸価とは、試料から抽出した油分の酸価をいう（公益社団法人日本油化学会制定の基準油脂分析試験法（日本油化学会規格試験法委員会編、基準油脂分析試験法（Ｓｔａｎｄａｒｄ ｍｅｔｈｏｄｓ ｆｏｒ ｔｈｅ ａｎａｌｙｓｉｓ ｏｆ ｆａｔｓ，ｏｉｌｓ ａｎｄ ｒｅｌａｔｅｄ ｍａｔｅｒｉａｌｓ）：日本油化学会制定、２０１３年版、１．５ 抽出油の酸価））。
<Acid value>
In the present invention, the acid value refers to the acid value of the oil extracted from the sample (standard oil and fat analysis test method established by the Japan Oil Chemicals Society (edited by the Japan Oil Chemicals Society Standard Test Method Committee, standard oil and fat analysis test). Law (Standard methods for the analysis of fats, oils and related materials): Established by the Japan Oil Chemists' Society, 2013 edition, 1.5 Acid value of extracted oil).
＜酸価の測定方法＞
本発明において、酸価の値は、公益社団法人日本油化学会制定の基準油脂分析試験法（日本油化学会規格試験法委員会編、基準油脂分析試験法（Ｓｔａｎｄａｒｄ ｍｅｔｈｏｄｓ ｆｏｒ ｔｈｅ ａｎａｌｙｓｉｓ ｏｆ ｆａｔｓ，ｏｉｌｓ ａｎｄ ｒｅｌａｔｅｄ ｍａｔｅｒｉａｌｓ）：日本油化学会制定、２０１３年版、１．５ 抽出油の酸価）により測定することができる。
<Measurement method of acid value>
In the present invention, the acid value is determined by the standard oil and fat analysis test method established by the Japan Oil Chemicals Society (edited by the Japan Oil Chemists' Society Standard Test Method Committee, Standard methods for the analysis of fats, oils and retarded materials): Established by the Japan Oil Chemists' Society, 2013 edition, 1.5 Acid value of extracted oil).
具体的には、まず、試料（リパーゼの固定化酵素を除いた反応液）をその推定酸価に対応する採取量に準じて三角フラスコに正しく測り取り、エタノール／ジエチルエーテル＝１／１（ｖ／ｖ）の混合溶媒１００ｍＬを加え、試料を完全に溶解させた。次いで、０．１ｍｏｌ／Ｌのエタノール性水酸化カリウムで滴定し、指示薬として加えたフェノールフタレイン溶液の変色が３０秒間以上続いた点を終点とする。酸価は以下の式（３）により算出された。
酸価＝５．６１１×Ａ×Ｆ／Ｂ ・・・・（３）
Ａ：０．１ｍｏｌ／Ｌのエタノール性水酸化カリウムの使用量（ｍＬ）
Ｂ：試料採取量（ｇ）
Ｆ：エタノール性水酸化カリウムのファクター
Specifically, first, the sample (reaction solution excluding the lipase-immobilized enzyme) was correctly measured in an Erlenmeyer flask according to the collected amount corresponding to the estimated acid value, and ethanol / diethyl ether = 1/1 (v). 100 mL of the mixed solvent of / v) was added to completely dissolve the sample. Next, titration with 0.1 mol / L ethanolic potassium hydroxide is performed, and the point at which the discoloration of the phenolphthalein solution added as an indicator continues for 30 seconds or longer is defined as the end point. The acid value was calculated by the following formula (3).
Acid value = 5.611 x A x F / B ... (3)
A: Amount of 0.1 mol / L ethanolic potassium hydroxide used (mL)
B: Sampling amount (g)
F: Factor of ethanolic potassium hydroxide
＜エステル交換反応＞
エステル交換反応とは通常、エステルにアルコールや酸、または他のエステルを作用させて、主鎖部分が入れ替わる反応を経て反応前とは別のエステルを生じる反応を指すが、本発明においてエステル交換反応とは、高度不飽和脂肪酸を含有する、モノグリセリド及びジグリセリドのエステルに対して基質となる遊離脂肪酸とリパーゼを作用させて、主鎖部分を入れ替える反応を指し、これにより高度不飽和脂肪酸及び中鎖脂肪酸含有トリグリセリドが製造される。
<Transesterification reaction>
The transesterification reaction usually refers to a reaction in which an alcohol, an acid, or another ester is allowed to act on an ester to exchange the main chain portion to produce an ester different from that before the reaction. In the present invention, the transesterification reaction is performed. Refers to a reaction in which a free fatty acid as a substrate and a lipase are allowed to act on an ester of a monoglyceride and a diglyceride containing a highly unsaturated fatty acid to replace the main chain portion, whereby the highly unsaturated fatty acid and the medium chain fatty acid are replaced. The contained triglyceride is produced.
＜エステル化反応＞
エステル化反応とは通常、カルボキシル基を持つカルボン酸類の化合物とヒドロキシル基を持つアルコール類またはフェノール類の化合物を触媒存在下で作用させて、エステルを生じる反応を指すが、本発明においてエステル化反応とは、高度不飽和脂肪酸を含有する、モノグリセリド及びジグリセリド中のヒドロキシル基に対して基質となる遊離脂肪酸とリパーゼを作用させて、新たにエステルを生じる反応を指し、これにより高度不飽和脂肪酸及び中鎖脂肪酸含有トリグリセリドが製造される。
本発明のエステル化反応は、副産物として水が生じる。反応系への水の共存は、エステル化反応の逆反応である加水分解反応を促進するため、可能な限り反応系内への水の混入を防ぐことや、副産物として生じた水を除きながらエステル化反応を進めるのが望ましい。
<Esterification reaction>
The esterification reaction usually refers to a reaction in which a compound of a carboxylic acid having a carboxyl group and a compound of an alcohol or a phenol having a hydroxyl group are allowed to act in the presence of a catalyst to produce an ester. In the present invention, the esterification reaction is performed. Refers to a reaction in which a free fatty acid as a substrate and a lipase are allowed to act on a hydroxyl group in a monoglyceride and a diglyceride containing a highly unsaturated fatty acid to form a new ester, whereby the highly unsaturated fatty acid and the medium are produced. Chain fatty acid-containing triglycerides are produced.
The esterification reaction of the present invention produces water as a by-product. Since the coexistence of water in the reaction system promotes the hydrolysis reaction, which is the reverse reaction of the esterification reaction, it is possible to prevent water from being mixed into the reaction system as much as possible, and ester while removing water generated as a by-product. It is desirable to proceed with the chemical reaction.
＜リパーゼ処理＞
本発明において、リパーゼ処理とは、例えば中鎖脂肪酸（特に遊離中鎖脂肪酸）のような基質と高度不飽和脂肪酸を含有する、モノグリセリド及びジグリセリドを、リパーゼと接触させることで、エステル交換反応やエステル化反応を進める処理のことを指す。
<Lipase treatment>
In the present invention, the lipase treatment is a transesterification reaction or ester by bringing a monoglyceride and a diglyceride containing a substrate such as a medium-chain fatty acid (particularly a free medium-chain fatty acid) and a highly unsaturated fatty acid into contact with lipase. Refers to the process of advancing the conversion reaction.
＜反応温度及び反応時間＞
本発明において、リパーゼ処理における反応液の温度は、用いるリパーゼの種類により適宜決定すればよいが、具体的には、例えば、２０℃以上６０℃以下とすることができ、さらに３０℃以上５０℃以下とすることができる。
<Reaction temperature and reaction time>
In the present invention, the temperature of the reaction solution in the lipase treatment may be appropriately determined depending on the type of lipase used, but specifically, for example, it can be 20 ° C. or higher and 60 ° C. or lower, and further 30 ° C. or higher and 50 ° C. It can be as follows.
また、反応時間は２時間以上とすることができ、さらに４時間以上とすることができ、さらに１６時間以上とすることができる。
反応時間の上限については特に制限はないが、効率的に反応させる観点から、４８時間以下であるとよく、さらに３６時間以下であるとよく、さらに２４時間以下であるとよく、特に２０時間以下であるとよい。
In addition, the reaction time can be 2 hours or more, further 4 hours or more, and further 16 hours or more.
The upper limit of the reaction time is not particularly limited, but from the viewpoint of efficient reaction, it is preferably 48 hours or less, 36 hours or less, 24 hours or less, and particularly 20 hours or less. It is good to be.
＜分子蒸留＞
本発明の製造方法において、グリセリドとそれ以外の成分に分離することができることからリパーゼ処理で得られた高度不飽和脂肪酸及び中鎖脂肪酸含有トリグリセリドを分子蒸留してもよい。
<Molecular distillation>
In the production method of the present invention, triglycerides containing polyunsaturated fatty acids and medium-chain fatty acids obtained by lipase treatment may be molecularly distilled because they can be separated into glycerides and other components.
分子蒸留に用いる装置は、一般的に市販されているものを使用して行うことができる。具体的には、例えば、遠心式分子蒸留機、ショートパス蒸留機、流下膜式蒸留機等を用いることができ、特に、ショートパス蒸留機を用いると良い。 The apparatus used for molecular distillation can be carried out by using a generally commercially available device. Specifically, for example, a centrifugal molecular distiller, a short pass distiller, a downflow membrane distiller, or the like can be used, and in particular, a short pass distiller is preferably used.
＜中鎖脂肪酸＞
本発明において、中鎖脂肪酸とは、構成脂肪酸が炭素数６〜１２、好ましくは８〜１２の中鎖脂肪酸である。
中鎖脂肪酸としては、具体的には、カプロン酸、カプリル酸、カプリン酸等が挙げられるが、中でも特に消化吸収に優れていることから構成脂肪酸の炭素数が８又は１０の中鎖脂肪酸がよく、特に炭素数が８の中鎖脂肪酸がよい。
また、中鎖脂肪酸の形態としては、遊離中鎖脂肪酸の形態の他、中鎖脂肪酸の低級アルコールエステルの形態等挙げられるが、高度不飽和脂肪酸を含有する、モノグリセリド及びジグリセリドから効率的に高度不飽和脂肪酸及び中鎖脂肪酸含有トリグリセリドを製造し易いことから、中鎖脂肪酸の形態としては遊離脂肪酸の形態であるとよい。
<Medium chain fatty acid>
In the present invention, the medium-chain fatty acid is a medium-chain fatty acid having 6 to 12 carbon atoms, preferably 8 to 12 carbon atoms.
Specific examples of the medium-chain fatty acid include caproic acid, caprylic acid, and capric acid. Among them, medium-chain fatty acids having 8 or 10 carbon atoms are often used because they are particularly excellent in digestion and absorption. In particular, medium-chain fatty acids having 8 carbon atoms are preferable.
Examples of the form of the medium-chain fatty acid include the form of a free medium-chain fatty acid and the form of a lower alcohol ester of the medium-chain fatty acid. However, the monoglyceride and the diglyceride containing the highly unsaturated fatty acid are efficiently highly unsaturated. Since it is easy to produce saturated fatty acids and triglycerides containing medium-chain fatty acids, the form of medium-chain fatty acids is preferably the form of free fatty acids.
＜高度不飽和脂肪酸を含有する、モノグリセリド及びジグリセリドに対する中鎖脂肪酸の量＞
本発明において、高度不飽和脂肪酸を含有する、モノグリセリド及びジグリセリドに対する中鎖脂肪酸の量は、前記中鎖脂肪酸が前記モノグリセリド及びジグリセリドの合計１００質量部に対し１０質量部以上１００質量部以下であるとよく、２０質量部以上８０質量部以下であればよく、特に２０質量部以上７０質量部以下であればよい。
<Amount of medium-chain fatty acids relative to monoglycerides and diglycerides containing highly unsaturated fatty acids>
In the present invention, the amount of the medium-chain fatty acid with respect to the monoglyceride and the diglyceride containing the highly unsaturated fatty acid is 10 parts by mass or more and 100 parts by mass or less based on 100 parts by mass of the total of the monoglyceride and the diglyceride. It may be 20 parts by mass or more and 80 parts by mass or less, and particularly 20 parts by mass or more and 70 parts by mass or less.
次に、本発明を実施例及び比較例に基づき、さらに説明する。
なお、本発明はこれに限定するものではない。
Next, the present invention will be further described based on Examples and Comparative Examples.
The present invention is not limited to this.
［実施例１］
高度不飽和脂肪酸を含有するモノグリセリド６．９４ｇ、高度不飽和脂肪酸を含有するジグリセリド１．５３ｇ、その他の成分を１．５３ｇ含む（前記モノグリセリド及びジグリセリドの質量比８２：１８。その他の成分の割合は、高度不飽和脂肪酸を含有する、モノグリセリド及びジグリセリドの合計１００質量部に対して１８質量部。）高度不飽和脂肪酸濃縮油脂（イワシ油由来）１０．０ｇと、遊離中鎖脂肪酸（カプリル酸；Ｃ８)３．５g をナスフラスコ(容量１００ｍL)に入れ、３０℃の恒温水槽上で回転させて均一化し、反応液を調製した。
なお、高度不飽和脂肪酸を含有する、モノグリセリド及びジグリセリドに含まれるDHA及びEPAの含有量は、それぞれ１９．６％、１７．２％であった。
また用いた中鎖脂肪酸は前記モノグリセリド及びジグリセリド１００質量部に対し４１質量部であった。
次に、上記反応液にリパーゼの固定化酵素（商品：Ｎｏｖｏｚｙｍ ４００８６（ノボザイムズ社製） ）を１．３５g添加し、リパーゼ処理を開始し、反応中は３０℃の恒温水槽上で回転させ、エバポレーター（真空度８０ｍｍHg）で反応系中を減圧しながら反応を進行させた。
反応開始から２１時間の時点での反応液を試料として採取した。
試料の採取は、反応系中のリパーゼの固定化酵素を除くため、ナスフラスコの回転を停止し、上澄み液のみを採取して試料とした。
得られた試料について、試料に対する酸価測定、脂質組成分析を行い、リパーゼ処理前後の高度不飽和脂肪酸及び中鎖脂肪酸含有トリグリセリドの増加量は、反応液（高度不飽和脂肪酸濃縮油脂及び遊離中鎖脂肪酸の計１３．５ｇ）に対し４３％（５．８g）であった。また、反応時間２１時間の試料に関して、脂肪酸組成分析を行い、高度不飽和脂肪酸及び中鎖脂肪酸含有トリグリセリド中に含まれる中鎖脂肪酸DHA及びEPAの含有量(質量%)を測定し、高度不飽和脂肪酸及び中鎖脂肪酸含有トリグリセリドに含まれる中鎖脂肪酸、DHA及びEPAの含有量は、それぞれ１９．７％、１７．３％、１５．３％であった。
なお、反応液の質量に対して、添加したリパーゼの固定化酵素は、担体の質量を含む質量として１０質量％であった。
[Example 1]
It contains 6.94 g of monoglyceride containing highly unsaturated fatty acid, 1.53 g of diglyceride containing highly unsaturated fatty acid, and 1.53 g of other components (mass ratio of monoglyceride and diglyceride 82:18. The ratio of other components is , 18 parts by mass with respect to 100 parts by mass of monoglyceride and diglyceride containing highly unsaturated fatty acids.) 10.0 g of highly unsaturated fatty acid concentrated fats and oils (derived from sardine oil) and free medium-chain fatty acids (capric acid; C8) ) 3.5 g was placed in a eggplant flask (capacity 100 mL) and rotated on a constant temperature water bath at 30 ° C. to homogenize, and a reaction solution was prepared.
The contents of DHA and EPA contained in monoglyceride and diglyceride containing highly unsaturated fatty acids were 19.6% and 17.2%, respectively.
The medium-chain fatty acid used was 41 parts by mass with respect to 100 parts by mass of the monoglyceride and diglyceride.
Next, 1.35 g of a lipase-immobilized enzyme (commodity: Novozyme 40086 (manufactured by Novozymes)) was added to the above reaction solution to start the lipase treatment, which was rotated on a constant temperature water bath at 30 ° C. during the reaction and evaporated. The reaction was carried out while reducing the pressure in the reaction system at (vacuum degree 80 mmHg).
The reaction solution at 21 hours from the start of the reaction was collected as a sample.
In order to remove the lipase-immobilized enzyme in the reaction system, the rotation of the eggplant flask was stopped and only the supernatant was collected as a sample.
For the obtained sample, acid value measurement and lipid composition analysis were performed on the sample, and the amount of increase in highly unsaturated fatty acid and medium-chain fatty acid-containing triglyceride before and after lipase treatment was determined by the reaction solution (highly unsaturated fatty acid concentrated fat and oil and free middle chain). It was 43% (5.8 g) with respect to 13.5 g of fatty acids in total. In addition, fatty acid composition analysis was performed on a sample with a reaction time of 21 hours, and the content (mass%) of medium-chain fatty acids DHA and EPA contained in highly unsaturated fatty acids and triglycerides containing medium-chain fatty acids was measured, and highly unsaturated fatty acids were measured. The contents of medium-chain fatty acid, DHA and EPA contained in the fatty acid and medium-chain fatty acid-containing triglyceride were 19.7%, 17.3% and 15.3%, respectively.
The amount of the lipase-immobilized enzyme added was 10% by mass including the mass of the carrier with respect to the mass of the reaction solution.
＜酸価測定＞
本発明における酸価の測定方法は ［００２８］に記載の方法で実施した。
酸価測定の結果と遊離中鎖脂肪酸の分子量から、試料中の遊離中鎖脂肪酸の含量(質量％)を算出した。
例えば、遊離中鎖脂肪酸にカプリル酸(C8FFA)を用いた場合、下記の式により算出できる。
［C8FFA含量(質量%)］＝［酸価］×144.21(C8FFAの分子量)÷56.11(KOHの分子量)÷10
<Acid value measurement>
The method for measuring the acid value in the present invention was carried out by the method described in [0028].
The content (mass%) of the free medium-chain fatty acid in the sample was calculated from the result of the acid value measurement and the molecular weight of the free medium-chain fatty acid.
For example, when caprylic acid (C8FFA) is used as the free medium-chain fatty acid, it can be calculated by the following formula.
[C8FFA content (mass%)] = [acid value] x 144.21 (molecular weight of C8FFA) ÷ 56.11 (molecular weight of KOH) ÷ 10
＜脂質組成分析(TLC-FID)＞
試料の１%(w/v) クロロホルム:メタノール混液（２：１、ｖ/ｖ）を調製して、薄層クロマトグラフィー検出装置（装置名：イアトロスキャン（登録商標） ＭＫ−６ｓ；LSIメディエンス社製)を用いたTLC-FID分析に供した。
展開はヘキサン：ジエチルエーテル：ギ酸（９５：５：０．１、ｖ/ｖ/ｖ）を溶媒に用いて行った。
当該脂質組成分析により、反応液中に存在する脂質組成を分析することができる。
また、酸価測定から得られた遊離中鎖脂肪酸の含量と組み合わせることで、脂質成分の種類毎(より具体的には、トリグリセリド、ジグリセリド、モノグリセリドや遊離中鎖脂肪酸)の含量を算出した。
<Lipid composition analysis (TLC-FID)>
Prepare a 1% (w / v) chloroform: methanol mixed solution (2: 1, v / v) of the sample and prepare a thin layer chromatography detector (device name: Iatroscan® MK-6s; LSI Medience). It was subjected to TLC-FID analysis using (manufactured by the company).
Development was carried out using hexane: diethyl ether: formic acid (95: 5: 0.1, v / v / v) as a solvent.
By the lipid composition analysis, the lipid composition present in the reaction solution can be analyzed.
In addition, the content of each type of lipid component (more specifically, triglyceride, diglyceride, monoglyceride, and free medium-chain fatty acid) was calculated by combining with the content of free medium-chain fatty acid obtained from the acid value measurement.
＜分取薄層クロマトグラフィー(ＴＬＣ)＞
試料を分取TLCにて脂質組成毎に分画後、トリグリセリド画分をかきとり、クロロホルム:メタノール混液（２：１、ｖ/ｖ）で抽出することで、試料中に含有するトリグリセリドを精製した。
具体的には、試料の5%(w/v)クロロホルム:メタノール混液（２：１、ｖ/ｖ）溶解液を調製し、分取用TLCに塗布して、クロロホルム:アセトン:酢酸（９０：１０：０．５、ｖ/ｖ/ｖ）の溶媒で展開後、前述のとおりクロロホルム:メタノール混液（２：１、ｖ/ｖ）でトリグリセリド画分を抽出し、脱溶媒することでトリグリセリド画分を精製した。
<Preparation Thin Layer Chromatography (TLC)>
After fractionating the sample by preparative TLC for each lipid composition, the triglyceride fraction was scraped off and extracted with a chloroform: methanol mixed solution (2: 1, v / v) to purify the triglyceride contained in the sample.
Specifically, a 5% (w / v) chloroform: methanol mixed solution (2: 1, v / v) solution of the sample was prepared, applied to a preparative TLC, and chloroform: acetone: acetic acid (90:). After developing with a solvent of 10: 0.5, v / v / v), the triglyceride fraction is extracted with a chloroform: methanol mixed solution (2: 1, v / v) as described above, and the triglyceride fraction is desolved to remove the solvent. Was purified.
＜脂肪酸組成分析＞
分取ＴＬＣにて精製したトリグリセリド画分をメチルエステル化後、ガスクロマトグラフィー（ＧＣ）分析することにより、トリグリセリド中に含有される脂肪酸組成を分析した。
メチルエステル化は、公益社団法人日本油化学会制定の基準油脂分析試験法(日本油化学会規格試験法委員会編、基準油脂分析試験法(Standard methods for the analysis of fats, oils and related materials):日本油化学会制定、2013年版、2.4.1.2 メチルエステル化(三フッ化ホウ素-メタノール法)）に従い行った。
メチルエステル化後、下記の条件にてGC分析に供した。
<Fatty acid composition analysis>
The triglyceride fraction purified by preparative TLC was methyl esterified and then analyzed by gas chromatography (GC) to analyze the fatty acid composition contained in the triglyceride.
Methyl esterification is a standard method for the analysis of fats, oils and related materials established by the Japan Oil Chemists'Association (edited by the Japan Oil Chemists' Society Standard Test Method Committee). : Established by Japan Oil Chemists' Society, 2013 edition, 2.4.1.2 Methyl esterification (boron trifluoride-methanol method)).
After methyl esterification, it was subjected to GC analysis under the following conditions.
カラム：Ｓｕｐｅｌｃｏ製 Ｏｍｅｇａｗａｘ２５０（Ｌ×Ｉ．Ｄ．３０ｍ×０．２５ｍｍ，ｄｆ:０．２５μｍ）
キャリアーガス：ヘリウム、カラム流量 １ｍＬ／ｍｉｎ
検出器：ＦＩＤ、２５０℃（水素:約３０ｍＬ／分、空気:約３００ｍＬ／分、窒素:約１８ｍＬ／分)
注入口：２５０℃、スプリット比（６０：１）
カラムオーブン温度：５０℃、２分；４℃／分，２２０℃（１５分）
装置：Ａｇｉｌｅｎｔ Ｔｅｃｈｎｏｌｏｇｉｅｓ製 ６８９０Ｎ
昇温条件：オーブン２０５℃、１０分保持 → 昇温２．５℃／分 → ２４０℃、１１分保持
分析時間：３５分
Column: Omegawax250 manufactured by Superco (L × ID 30 m × 0.25 mm, df: 0.25 μm)
Carrier gas: helium, column flow rate 1 mL / min
Detector: FID, 250 ° C (hydrogen: about 30 mL / min, air: about 300 mL / min, nitrogen: about 18 mL / min)
Injection port: 250 ° C, split ratio (60: 1)
Column oven temperature: 50 ° C, 2 minutes; 4 ° C / min, 220 ° C (15 minutes)
Equipment: Agilent Technologies 6890N
Temperature rise conditions: Oven 205 ° C, 10 minutes hold → Temperature rise 2.5 ° C / min → 240 ° C, 11 minutes hold Analysis time: 35 minutes
［実施例２］
実施例１から高度不飽和脂肪酸を含有するモノグリセリドと高度不飽和脂肪酸を含有するジグリセリドの割合が異なる油脂に変更した。
具体的には、高度不飽和脂肪酸を含有するモノグリセリド１．８４ｇ、高度不飽和脂肪酸を含有するジグリセリド６．６１ｇ、その他の成分を１．５５ｇ含む（前記モノグリセリド及びジグリセリドの質量比２２：７８。その他の成分の割合は、高度不飽和脂肪酸を含有する、モノグリセリド及びジグリセリドの合計１００質量部に対して１８質量部。）高度不飽和脂肪酸濃縮油脂（イワシ油由来）１０．０ｇを用いた。
それ以外は、実施例１と同様の方法で実施例２の反応液の調製および反応の進行を行った。
なお、高度不飽和脂肪酸を含有する、モノグリセリド及びジグリセリドに含まれるDHA及びEPAの含有量は、それぞれ２３．２％、１６．４％であった。
また用いた中鎖脂肪酸は前記モノグリセリド及びジグリセリド１００質量部に対し４１質量部であった。
さらに、反応液の質量に対して、添加したリパーゼの固定化酵素は、担体の質量を含む質量として１０質量％であった。
得られた試料について、試料に対する酸価測定、脂質組成分析、脂肪酸組成分析を行い、リパーゼ処理前後の高度不飽和脂肪酸及び中鎖脂肪酸含有トリグリセリドの増加量は、反応液（高度不飽和脂肪酸濃縮油脂及び遊離中鎖脂肪酸の計１３．５ｇ）に対し５１％（６．９g）、高度不飽和脂肪酸及び中鎖脂肪酸含有トリグリセリド中の中鎖脂肪酸の含有量１９．１％を求めた。
また、得られた試料において高度不飽和脂肪酸及び中鎖脂肪酸含有トリグリセリドに含まれるDHA及びEPAの含有量は、それぞれ１９．１％、１２．９％であった。
[Example 2]
From Example 1, the ratio of monoglyceride containing highly unsaturated fatty acid and diglyceride containing highly unsaturated fatty acid was changed to a different fat and oil.
Specifically, it contains 1.84 g of monoglyceride containing highly unsaturated fatty acid, 6.61 g of diglyceride containing highly unsaturated fatty acid, and 1.55 g of other components (mass ratio of monoglyceride to diglyceride 22:78. Others. The ratio of the components of was 18 parts by mass with respect to 100 parts by mass of the total of monoglyceride and diglyceride containing highly unsaturated fatty acids.) 10.0 g of highly unsaturated fatty acid concentrated fats and oils (derived from sardine oil) was used.
Other than that, the reaction solution of Example 2 was prepared and the reaction proceeded in the same manner as in Example 1.
The contents of DHA and EPA contained in monoglyceride and diglyceride containing highly unsaturated fatty acids were 23.2% and 16.4%, respectively.
The medium-chain fatty acid used was 41 parts by mass with respect to 100 parts by mass of the monoglyceride and diglyceride.
Further, the amount of the added lipase immobilized enzyme was 10% by mass including the mass of the carrier with respect to the mass of the reaction solution.
Acid value measurement, lipid composition analysis, and fatty acid composition analysis were performed on the obtained sample, and the amount of increase in highly unsaturated fatty acid and medium-chain fatty acid-containing triglyceride before and after lipase treatment was determined by the reaction solution (highly unsaturated fatty acid concentrated fat and oil). And 51% (6.9 g) of free medium-chain fatty acid (13.5 g in total), and 19.1% of medium-chain fatty acid content in highly unsaturated fatty acid and medium-chain fatty acid-containing triglyceride was determined.
In the obtained sample, the contents of DHA and EPA contained in the polyunsaturated fatty acid and the medium chain fatty acid-containing triglyceride were 19.1% and 12.9%, respectively.
［比較例１］
実施例１から高度不飽和脂肪酸を含有するモノグリセリドと高度不飽和脂肪酸を含有するジグリセリドの割合が異なる油脂に変更した。
具体的には、高度不飽和脂肪酸を含有するモノグリセリド０．５４ｇと高度不飽和脂肪酸を含有するジグリセリド７．９０ｇを含む（前記モノグリセリド及びジグリセリドの質量比６：９４。その他の成分の割合は、高度不飽和脂肪酸を含有する、モノグリセリド及びジグリセリドの合計１００質量部に対して１８質量部。）高度不飽和脂肪酸濃縮油脂（イワシ油由来）１０．０ｇとした。
それ以外は、実施例１と同様の方法で比較例１の反応液の調製および反応の進行を行った。
なお、高度不飽和脂肪酸を含有する、モノグリセリド及びジグリセリドに含まれるDHA及びEPAの含有量は、それぞれ２４．２％、１６．１％であった。
また用いた遊離中鎖脂肪酸は前記モノグリセリド及びジグリセリド１００質量部に対し４１質量部であった。
さらに、反応液の質量に対して、添加したリパーゼの固定化酵素は、担体の質量を含む質量として１０質量％であった。
得られた試料について、試料に対する酸価測定、脂質組成分析、脂肪酸組成分析を行い、リパーゼ処理前後の高度不飽和脂肪酸及び中鎖脂肪酸含有トリグリセリドの増加量は、反応液（高度不飽和脂肪酸濃縮油脂及び遊離中鎖脂肪酸の計１３．５ｇ）に対し４５％（６．１g）、高度不飽和脂肪酸及び中鎖脂肪酸含有トリグリセリド中の中鎖脂肪酸の含有量１６．５％を求めた。
得られた試料において高度不飽和脂肪酸及び中鎖脂肪酸含有トリグリセリドに含まれるDHA及びEPAの含有量は、それぞれ２０．３％、１２．５％であった。
[Comparative Example 1]
From Example 1, the ratio of monoglyceride containing highly unsaturated fatty acid and diglyceride containing highly unsaturated fatty acid was changed to a different fat and oil.
Specifically, it contains 0.54 g of monoglyceride containing highly unsaturated fatty acid and 7.90 g of diglyceride containing highly unsaturated fatty acid (mass ratio of monoglyceride to diglyceride is 6:94. The ratio of other components is high. 18 parts by mass with respect to 100 parts by mass of the total of monoglyceride and diglyceride containing unsaturated fatty acids.) 10.0 g of highly unsaturated fatty acid concentrated fats and oils (derived from sardine oil) was used.
Other than that, the reaction solution of Comparative Example 1 was prepared and the reaction proceeded in the same manner as in Example 1.
The contents of DHA and EPA contained in monoglyceride and diglyceride containing highly unsaturated fatty acids were 24.2% and 16.1%, respectively.
The amount of free medium-chain fatty acid used was 41 parts by mass with respect to 100 parts by mass of the monoglyceride and diglyceride.
Further, the amount of the added lipase immobilized enzyme was 10% by mass including the mass of the carrier with respect to the mass of the reaction solution.
Acid value measurement, lipid composition analysis, and fatty acid composition analysis were performed on the obtained sample, and the amount of increase in highly unsaturated fatty acid and medium-chain fatty acid-containing triglyceride before and after lipase treatment was determined by the reaction solution (highly unsaturated fatty acid concentrated fat and oil). And 45% (6.1 g) of free medium-chain fatty acids (13.5 g in total), and the content of medium-chain fatty acids in highly unsaturated fatty acids and triglycerides containing medium-chain fatty acids was determined to be 16.5%.
In the obtained sample, the contents of DHA and EPA contained in the polyunsaturated fatty acid and the medium chain fatty acid-containing triglyceride were 20.3% and 12.5%, respectively.
実施例１、実施例２および比較例１から得られた結果を表１に示した。
［表１］
[Table 1]
なお、表１において、「反応液に対するリパーゼ処理前後の高度不飽和脂肪酸及び中鎖脂肪酸含有トリグリセリドの増加量(質量％)とその評価」は以下の通りである。
◎：反応液に対するリパーゼ処理前後の高度不飽和脂肪酸及び中鎖脂肪酸含有トリグリセリドの増加量が５０％以上であり、高度不飽和脂肪酸及び中鎖脂肪酸含有トリグリセリドの生産効率に大変優れている。
〇：反応液に対するリパーゼ処理前後の高度不飽和脂肪酸及び中鎖脂肪酸含有トリグリセリドの増加量が４０％以上５０％未満であり、高度不飽和脂肪酸及び中鎖脂肪酸含有トリグリセリドの生産効率に優れている。
×：反応液に対するリパーゼ処理前後の高度不飽和脂肪酸及び中鎖脂肪酸含有トリグリセリドの増加量が４０％未満であり、高度不飽和脂肪酸及び中鎖脂肪酸含有トリグリセリドの生産効率に優れない。
In Table 1, "increase in polyunsaturated fatty acid and medium chain fatty acid-containing triglyceride (mass%) before and after lipase treatment of the reaction solution and its evaluation" are as follows.
⊚: The amount of increase in highly unsaturated fatty acid and medium chain fatty acid-containing triglyceride before and after lipase treatment with respect to the reaction solution is 50% or more, and the production efficiency of highly unsaturated fatty acid and medium chain fatty acid-containing triglyceride is very excellent.
〇: The amount of increase in highly unsaturated fatty acid and medium-chain fatty acid-containing triglyceride before and after lipase treatment with respect to the reaction solution is 40% or more and less than 50%, and the production efficiency of highly unsaturated fatty acid and medium-chain fatty acid-containing triglyceride is excellent.
X: The amount of increase in highly unsaturated fatty acid and medium-chain fatty acid-containing triglyceride before and after lipase treatment with respect to the reaction solution is less than 40%, and the production efficiency of highly unsaturated fatty acid and medium-chain fatty acid-containing triglyceride is not excellent.
表１において、「高度不飽和脂肪酸及び中鎖脂肪酸含有トリグリセリド中の中鎖脂肪酸の含有量(質量％)とその評価」は以下の通りである。
◎：高度不飽和脂肪酸及び中鎖脂肪酸含有トリグリセリド中の中鎖脂肪酸の含有量が１９%以上であり、高度不飽和脂肪酸及び中鎖脂肪酸の含有量が大変優れている。
〇：高度不飽和脂肪酸及び中鎖脂肪酸含有トリグリセリド中の中鎖脂肪酸の含有量が１７%以上１９%未満であり、高度不飽和脂肪酸及び中鎖脂肪酸の含有量が優れている。
△：高度不飽和脂肪酸及び中鎖脂肪酸含有トリグリセリド中の中鎖脂肪酸の含有量が１７%未満であり、高度不飽和脂肪酸及び中鎖脂肪酸の含有量に優れない。
In Table 1, "content (% by mass) of medium-chain fatty acid in polyunsaturated fatty acid and medium-chain fatty acid-containing triglyceride and its evaluation" are as follows.
⊚: Highly unsaturated fatty acid and medium chain fatty acid content The content of medium chain fatty acid in the triglyceride is 19% or more, and the content of highly unsaturated fatty acid and medium chain fatty acid is very excellent.
〇: Highly unsaturated fatty acid and medium chain fatty acid content The content of medium chain fatty acid in the triglyceride is 17% or more and less than 19%, and the content of highly unsaturated fatty acid and medium chain fatty acid is excellent.
Δ: The content of the medium-chain fatty acid in the highly unsaturated fatty acid and the medium-chain fatty acid-containing triglyceride is less than 17%, and the content of the highly unsaturated fatty acid and the medium-chain fatty acid is not excellent.
表１において、比較例１の［反応液に対するリパーゼ処理前後の高度不飽和脂肪酸及び中鎖脂肪酸含有トリグリセリドの増加量(％)］×［高度不飽和脂肪酸及び中鎖脂肪酸含有トリグリセリド中の中鎖脂肪酸の含有量(％)］の値、つまり比較例１のリパーゼ処理前後により増加した高度不飽和脂肪酸及び中鎖脂肪酸含有トリグリセリドの中の中鎖脂肪酸の含有量を１００（基準）とし、実施例１及び実施例２の、［反応液に対するリパーゼ処理前後の高度不飽和脂肪酸及び中鎖脂肪酸含有トリグリセリドの増加量(％)］×［高度不飽和脂肪酸及び中鎖脂肪酸含有トリグリセリド中の中鎖脂肪酸の含有量(％)］を求めたところ、それぞれ１１５、１３１となった。 In Table 1, [Increase in highly unsaturated fatty acid and medium-chain fatty acid-containing triglyceride (%) before and after lipase treatment with respect to the reaction solution] × [Medium-chain fatty acid in highly unsaturated fatty acid and medium-chain fatty acid-containing triglyceride] of Comparative Example 1. Content (%)], that is, the content of medium-chain fatty acids in the highly unsaturated fatty acid and medium-chain fatty acid-containing triglyceride that increased before and after the lipase treatment of Comparative Example 1 was set to 100 (reference), and Example 1 And Example 2, [Increase in highly unsaturated fatty acid and medium-chain fatty acid-containing triglyceride before and after lipase treatment with respect to the reaction solution (%)] × [Containance of medium-chain fatty acid in highly unsaturated fatty acid and medium-chain fatty acid-containing triglyceride] Amount (%)] was calculated and found to be 115 and 131, respectively.
表１において、「総合評価」は以下の通りである。
◎：比較例１に比べ、高度不飽和脂肪酸及び中鎖脂肪酸含有トリグリセリドの生産効率に大変優れており、かつ、高度不飽和脂肪酸及び中鎖脂肪酸の含有量にも大変優れている。
〇：比較例１に比べ、高度不飽和脂肪酸及び中鎖脂肪酸含有トリグリセリドの生産効率に優れており、かつ、高度不飽和脂肪酸及び中鎖脂肪酸の含有量にも優れている。
△：高度不飽和脂肪酸及び中鎖脂肪酸含有トリグリセリドの生産効率が優れていない、若しくは、高度不飽和脂肪酸及び中鎖脂肪酸の含有量に優れない。
In Table 1, the "comprehensive evaluation" is as follows.
⊚: Compared with Comparative Example 1, the production efficiency of highly unsaturated fatty acid and medium chain fatty acid-containing triglyceride is very excellent, and the content of highly unsaturated fatty acid and medium chain fatty acid is also very excellent.
〇: Compared with Comparative Example 1, the production efficiency of polyunsaturated fatty acid and medium chain fatty acid-containing triglyceride is excellent, and the content of polyunsaturated fatty acid and medium chain fatty acid is also excellent.
Δ: The production efficiency of highly unsaturated fatty acid and medium chain fatty acid-containing triglyceride is not excellent, or the content of highly unsaturated fatty acid and medium chain fatty acid is not excellent.
以上のことから、用いる高度不飽和脂肪酸を含有する、モノグリセリド及びジグリセリド
の質量比が２０：８０〜８５：１５であると、高度不飽和脂肪酸及び中鎖脂肪酸含有トリグリセリドの生産効率に優れており、かつ高度不飽和脂肪酸及び中鎖脂肪酸の含有量にも大変優れていることが明らかとなった。
From the above, when the mass ratio of monoglyceride and diglyceride containing highly unsaturated fatty acid to be used is 20:80 to 85:15, the production efficiency of highly unsaturated fatty acid and triglyceride containing medium-chain fatty acid is excellent. Moreover, it was revealed that the contents of highly unsaturated fatty acids and medium-chain fatty acids are also very excellent.
［実施例３］
高度不飽和脂肪酸を含有するモノグリセリド２．３２ｇ、高度不飽和脂肪酸を含有するジグリセリド５．６１ｇ、その他の成分を２．０７ｇ含む（前記モノグリセリド及びジグリセリドの質量比２９：７１。その他の成分の割合は、高度不飽和脂肪酸を含有する、モノグリセリド及びジグリセリドに対して２６質量部。）高度不飽和脂肪酸濃縮油脂（イワシ油由来）１０．０ｇと、遊離中鎖脂肪酸（カプリル酸；Ｃ８)２．０g をナスフラスコ(容量１００ｍL)に入れ、３０℃の恒温水槽上で回転させて均一化し、反応液を調製した。
なお、高度不飽和脂肪酸を含有する、モノグリセリド及びジグリセリドに含まれるDHA及びEPAの含有量は、それぞれ２４．６％、１７．７％であった。
また用いた中鎖脂肪酸は前記モノグリセリド及びジグリセリド１００質量部に対し２５質量部であった。
次に、上記反応液にリパーゼの固定化酵素（商品名：Ｌｉｐｏｚｙｍｅ（登録商標） ＲＭＩＭ（ノボザイムズ社製））を１．２０ｇ添加し、リパーゼ処理を開始した。
反応中は３０℃の恒温水槽上で回転させ、エバポレーター(真空度２０〜３０ｍｍＨｇ)で反応系中を減圧しながら反応を進行させた。
反応開始から０、２、４、６、１６、２４時間の時点で試料(反応液)を少量採取した。試料の採取は、反応系中のリパーゼの固定化酵素を除くため、ナスフラスコの回転を停止し、上澄み液のみを採取して試料とした。得られた試料に対して、実施例１と同様に、酸価測定、脂質組成分析を行い、反応の進行の指標として「反応液に対するリパーゼ処理前後の高度不飽和脂肪酸及び中鎖脂肪酸含有トリグリセリドの増加量(％)の評価」を実施した。
リパーゼ処理の反応時間は２時間以上とすると、反応液に対するリパーゼ処理前後の高度不飽和脂肪酸及び中鎖脂肪酸含有トリグリセリドの増加量が４０％以上５０％未満であり、高度不飽和脂肪酸及び中鎖脂肪酸含有トリグリセリドの生産効率に優れており、さらに４時間以上とすると反応液に対するリパーゼ処理前後の高度不飽和脂肪酸及び中鎖脂肪酸含有トリグリセリドの増加量が５０％以上であり、高度不飽和脂肪酸及び中鎖脂肪酸含有トリグリセリドの生産効率に大変優れていることが明らかとなった。
なお、反応液の質量に対して、添加したリパーゼの固定化酵素は、担体の質量を含む質量として１０質量％であった。
また、２、４、６、１６、２４時間の時点で、高度不飽和脂肪酸及び中鎖脂肪酸含有トリグリセリドに含まれるDHA及びEPAの含有量は、それぞれ１０％以上４０％以下、５％以上３５％以下であった。
[Example 3]
It contains 2.32 g of monoglyceride containing highly unsaturated fatty acid, 5.61 g of diglyceride containing highly unsaturated fatty acid, and 2.07 g of other components (mass ratio of monoglyceride and diglyceride 29:71. The ratio of other components is , 26 parts by mass with respect to monoglyceride and diglyceride containing highly unsaturated fatty acid.) 10.0 g of highly unsaturated fatty acid concentrated fat (derived from sardine oil) and 2.0 g of free medium chain fatty acid (capric acid; C8) It was placed in a eggplant flask (capacity 100 mL) and rotated on a constant temperature water bath at 30 ° C. to homogenize it, and a reaction solution was prepared.
The contents of DHA and EPA contained in monoglyceride and diglyceride containing highly unsaturated fatty acids were 24.6% and 17.7%, respectively.
The medium-chain fatty acid used was 25 parts by mass with respect to 100 parts by mass of the monoglyceride and diglyceride.
Next, 1.20 g of a lipase-immobilized enzyme (trade name: Lipozyme (registered trademark) RMIM (manufactured by Novozymes)) was added to the above reaction solution, and the lipase treatment was started.
During the reaction, the reaction was rotated on a constant temperature water bath at 30 ° C., and the reaction was carried out while reducing the pressure in the reaction system with an evaporator (vacuum degree 20 to 30 mmHg).
A small amount of sample (reaction solution) was collected at 0, 2, 4, 6, 16, and 24 hours from the start of the reaction. In order to remove the lipase-immobilized enzyme in the reaction system, the rotation of the eggplant flask was stopped and only the supernatant was collected as a sample. Acid value measurement and lipid composition analysis were performed on the obtained sample in the same manner as in Example 1, and as an index of the progress of the reaction, "highly unsaturated fatty acids and medium-chain fatty acid-containing triglycerides before and after lipase treatment of the reaction solution" were used. Evaluation of the amount of increase (%) ”was carried out.
When the reaction time of the lipase treatment is 2 hours or more, the amount of increase in highly unsaturated fatty acid and medium chain fatty acid-containing triglyceride before and after the lipase treatment with respect to the reaction solution is 40% or more and less than 50%, and the highly unsaturated fatty acid and medium chain fatty acid. The production efficiency of the contained triglyceride is excellent, and when it is further set to 4 hours or more, the increase amount of the highly unsaturated fatty acid and the medium chain fatty acid-containing triglyceride before and after the lipase treatment with respect to the reaction solution is 50% or more, and the highly unsaturated fatty acid and the medium chain It was revealed that the production efficiency of fatty acid-containing triglyceride is very excellent.
The amount of the lipase-immobilized enzyme added was 10% by mass including the mass of the carrier with respect to the mass of the reaction solution.
At 2, 4, 6, 16 and 24 hours, the contents of DHA and EPA contained in the polyunsaturated fatty acid and medium chain fatty acid-containing triglyceride were 10% or more and 40% or less, and 5% or more and 35%, respectively. It was as follows.
［実施例４］
実施例３から、遊離中鎖脂肪酸（カプリル酸；Ｃ８)の量を３．５ｇ、リパーゼの固定化酵素（商品名：Ｌｉｐｏｚｙｍｅ ＲＭＩＭ（ノボザイムズ社製））の量を１．３５ｇに変更した。それ以外は実施例３と同様の方法で実施例４の反応液の調製および反応の進行を行った。試料の採取を、反応開始から０、２、４、６、１２、１６、２０、２４時間の時点で試料(反応液)を少量採取し、実施例１と同様に、酸価測定、脂質組成分析を行い、反応の進行の指標として「反応液に対するリパーゼ処理前後の高度不飽和脂肪酸及び中鎖脂肪酸含有トリグリセリドの増加量(質量％)の評価」を実施した。
リパーゼ処理の反応時間は２時間以上とすると、反応液に対するリパーゼ処理前後の高度不飽和脂肪酸及び中鎖脂肪酸含有トリグリセリドの増加量が４０％以上５０％未満であり、高度不飽和脂肪酸及び中鎖脂肪酸含有トリグリセリドの生産効率に優れており、さらに４時間以上とすると反応液に対するリパーゼ処理前後の高度不飽和脂肪酸及び中鎖脂肪酸含有トリグリセリドの増加量が５０％以上であり、高度不飽和脂肪酸及び中鎖脂肪酸含有トリグリセリドの生産効率に大変優れていることが明らかとなった。
用いた中鎖脂肪酸は前記モノグリセリド及びジグリセリド１００質量部に対し４１質量部であった。
なお、反応液の質量に対して、添加したリパーゼの固定化酵素は、担体の質量を含む質量として１０質量％であった。
また、２、４、６、１２、１６、２０、２４時間の時点で、高度不飽和脂肪酸及び中鎖脂肪酸含有トリグリセリドに含まれるDHA及びEPAの含有量は、それぞれ１０％以上４０％以下、５％以上３５％以下であった。
[Example 4]
From Example 3, the amount of free medium-chain fatty acid (caprylic acid; C8) was changed to 3.5 g, and the amount of lipase immobilized enzyme (trade name: Lipozyme RMIM (manufactured by Novozymes)) was changed to 1.35 g. Other than that, the reaction solution of Example 4 was prepared and the reaction proceeded in the same manner as in Example 3. A small amount of sample (reaction solution) was collected at 0, 2, 4, 6, 12, 16, 20, and 24 hours from the start of the reaction, and the acid value was measured and the lipid composition was measured in the same manner as in Example 1. The analysis was carried out, and "evaluation of the amount of increase (mass%) of highly unsaturated fatty acid and medium-chain fatty acid-containing triglyceride before and after lipase treatment on the reaction solution" was carried out as an index of the progress of the reaction.
When the reaction time of the lipase treatment is 2 hours or more, the amount of increase in highly unsaturated fatty acid and medium chain fatty acid-containing triglyceride before and after the lipase treatment with respect to the reaction solution is 40% or more and less than 50%, and the highly unsaturated fatty acid and medium chain fatty acid. The production efficiency of the contained triglyceride is excellent, and when it is further set to 4 hours or more, the increase amount of the highly unsaturated fatty acid and the medium chain fatty acid-containing triglyceride before and after the lipase treatment with respect to the reaction solution is 50% or more, and the highly unsaturated fatty acid and the medium chain It was revealed that the production efficiency of fatty acid-containing triglyceride is very excellent.
The medium-chain fatty acid used was 41 parts by mass with respect to 100 parts by mass of the monoglyceride and diglyceride.
The amount of the lipase-immobilized enzyme added was 10% by mass including the mass of the carrier with respect to the mass of the reaction solution.
At 2, 4, 6, 12, 16, 20, and 24 hours, the contents of DHA and EPA contained in the polyunsaturated fatty acid and medium chain fatty acid-containing triglyceride were 10% or more and 40% or less, respectively, 5 It was% or more and 35% or less.
［実施例５］
実施例３から、遊離中鎖脂肪酸(カプリル酸；Ｃ８）の量を５．０ｇ、リパーゼの固定化酵素であるＬｉｐｏｚｙｍｅ ＲＭＩＭ（ノボザイムズ社製）の量を１．５ｇに変更した。それ以外は実施例３，４と同様の方法で実施例５の反応液の調製および反応の進行を行った。試料の採取を、反応開始から０、２、４、６、１２、１６、２０、２４時間の時点で試料(反応液)を少量採取し、実施例１と同様に、酸価測定、脂質組成分析を行い、反応の進行の指標として「反応液に対するリパーゼ処理前後の高度不飽和脂肪酸及び中鎖脂肪酸含有トリグリセリドの増加量(質量％)の評価」を実施した。
リパーゼ処理の反応時間は２時間以上とすると、反応液に対するリパーゼ処理前後の高度不飽和脂肪酸及び中鎖脂肪酸含有トリグリセリドの増加量が４０％以上５０％未満であり、高度不飽和脂肪酸及び中鎖脂肪酸含有トリグリセリドの生産効率に優れており、さらに４時間以上とすると反応液に対するリパーゼ処理前後の高度不飽和脂肪酸及び中鎖脂肪酸含有トリグリセリドの増加量が５０％以上であり、高度不飽和脂肪酸及び中鎖脂肪酸含有トリグリセリドの生産効率に大変優れていることが明らかとなった。
用いた中鎖脂肪酸は前記モノグリセリド及びジグリセリド１００質量部に対し６３質量部であった。
なお、反応液の質量に対して、添加したリパーゼの固定化酵素は、担体の質量を含む質量として１０質量％であり、高度不飽和脂肪酸及び中鎖脂肪酸含有トリグリセリドに含まれるDHA及びEPAの含有量は、それぞれ１０％以上４０％以下、５％以上３５％以下であった。
[Example 5]
From Example 3, the amount of free medium-chain fatty acid (caprylic acid; C8) was changed to 5.0 g, and the amount of lipase-immobilized enzyme Lipozyme RMIM (manufactured by Novozymes) was changed to 1.5 g. Other than that, the reaction solution of Example 5 was prepared and the reaction proceeded in the same manner as in Examples 3 and 4. A small amount of sample (reaction solution) was collected at 0, 2, 4, 6, 12, 16, 20, and 24 hours from the start of the reaction, and the acid value was measured and the lipid composition was measured in the same manner as in Example 1. The analysis was carried out, and "evaluation of the amount of increase (mass%) of highly unsaturated fatty acid and medium-chain fatty acid-containing triglyceride before and after lipase treatment on the reaction solution" was carried out as an index of the progress of the reaction.
When the reaction time of the lipase treatment is 2 hours or more, the amount of increase in highly unsaturated fatty acid and medium chain fatty acid-containing triglyceride before and after the lipase treatment with respect to the reaction solution is 40% or more and less than 50%, and the highly unsaturated fatty acid and medium chain fatty acid. The production efficiency of the contained triglyceride is excellent, and when it is further set to 4 hours or more, the increase amount of the highly unsaturated fatty acid and the medium chain fatty acid-containing triglyceride before and after the lipase treatment with respect to the reaction solution is 50% or more, and the highly unsaturated fatty acid and the medium chain It was revealed that the production efficiency of fatty acid-containing triglyceride is very excellent.
The medium-chain fatty acid used was 63 parts by mass with respect to 100 parts by mass of the monoglyceride and diglyceride.
The added lipase-immobilized enzyme has a mass of 10% by mass including the mass of the carrier with respect to the mass of the reaction solution, and contains DHA and EPA contained in the polyunsaturated fatty acid and the medium-chain fatty acid-containing triglyceride. The amounts were 10% or more and 40% or less, and 5% or more and 35% or less, respectively.
［表３］反応時間毎の反応液に対するリパーゼ処理前後の高度不飽和脂肪酸及び中鎖脂肪酸含有トリグリセリドの増加量(質量％)
以上のことから、リパーゼ処理の反応時間は２時間以上とするとよく、さらに４時間以上とするとよく、さらに１６時間以上とするとよいことが明らかとなった。
また、高度不飽和脂肪酸を含有する、モノグリセリド及びジグリセリドに対する中鎖脂肪酸の量は、前記中鎖脂肪酸が前記モノグリセリド及びジグリセリドの合計１００質量部に対し、１０質量部以上１００質量部以下であるとよく、２０質量部以上８０質量部以下であればよく、特に２０質量部以上７０質量部以下であればよいことが明らかとなった。
From the above, it was clarified that the reaction time of the lipase treatment should be 2 hours or more, further 4 hours or more, and further 16 hours or more.
The amount of medium-chain fatty acid with respect to monoglyceride and diglyceride containing highly unsaturated fatty acid is preferably 10 parts by mass or more and 100 parts by mass or less with respect to 100 parts by mass of the total of the monoglyceride and diglyceride. It has been clarified that the amount may be 20 parts by mass or more and 80 parts by mass or less, and particularly 20 parts by mass or more and 70 parts by mass or less.
［実施例６］
実施例３から、恒温水槽の温度を５０℃に変更し、それ以外は実施例３と同様の方法で実施例６の反応液の調製および反応の進行を行った。試料の採取を、反応開始から０、２、４、６、８、１６、２０、２４時間の時点で試料(反応液)を少量採取し、実施例１と同様に、酸価測定、脂質組成分析を行い、反応の進行の指標として「反応液に対するリパーゼ処理前後の高度不飽和脂肪酸及び中鎖脂肪酸含有トリグリセリドの増加量(質量％)の評価」を実施した。
[Example 6]
From Example 3, the temperature of the constant temperature water tank was changed to 50 ° C., and the reaction solution of Example 6 was prepared and the reaction proceeded in the same manner as in Example 3 except for the above. A small amount of sample (reaction solution) was collected at 0, 2, 4, 6, 8, 16, 20, and 24 hours from the start of the reaction, and the acid value was measured and the lipid composition was measured in the same manner as in Example 1. The analysis was carried out, and "evaluation of the amount of increase (mass%) of highly unsaturated fatty acid and medium-chain fatty acid-containing triglyceride before and after lipase treatment on the reaction solution" was carried out as an index of the progress of the reaction.
［表４］反応時間毎の反応液に対するリパーゼ処理前後の高度不飽和脂肪酸及び中鎖脂肪酸含有トリグリセリドの増加量(質量％)
以上のことから、リパーゼ処理における反応液の温度は、２０℃以上６０℃以下とすることができ、さらに３０℃以上５０℃以下とすることができることが明らかとなった。
From the above, it was clarified that the temperature of the reaction solution in the lipase treatment can be 20 ° C. or higher and 60 ° C. or lower, and further 30 ° C. or higher and 50 ° C. or lower.
Claims (2)
前記モノグリセリド及びジグリセリドの質量比が２０：８０〜８５：１５であることを特徴とする、
高度不飽和脂肪酸及び中鎖脂肪酸含有トリグリセリドの製造方法。 A method for producing a highly unsaturated fatty acid and a medium-chain fatty acid-containing triglyceride, in which a monoglyceride and a diglyceride containing a highly unsaturated fatty acid are lipase-treated in the presence of a medium-chain fatty acid.
The mass ratio of the monoglyceride to the diglyceride is 20:80 to 85:15.
A method for producing a triglyceride containing a polyunsaturated fatty acid and a medium chain fatty acid.
請求項１記載の高度不飽和脂肪酸及び中鎖脂肪酸含有トリグリセリドの製造方法。 The medium-chain fatty acid is 10 parts by mass or more and 100 parts by mass or less with respect to 100 parts by mass in total of the monoglyceride and diglyceride.
The method for producing a polyunsaturated fatty acid and a medium chain fatty acid-containing triglyceride according to claim 1.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP2019078911A JP2020174570A (en) | 2019-04-17 | 2019-04-17 | Method for producing polyunsaturated fatty acid and medium chain fatty acid-containing triglyceride |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP2019078911A JP2020174570A (en) | 2019-04-17 | 2019-04-17 | Method for producing polyunsaturated fatty acid and medium chain fatty acid-containing triglyceride |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| JP2020174570A true JP2020174570A (en) | 2020-10-29 |
Family
ID=72935383
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP2019078911A Pending JP2020174570A (en) | 2019-04-17 | 2019-04-17 | Method for producing polyunsaturated fatty acid and medium chain fatty acid-containing triglyceride |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JP2020174570A (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114480518A (en) * | 2022-02-28 | 2022-05-13 | 江南大学 | Method for preparing medium-long carbon chain triglyceride by enzyme method |
| CN114891568A (en) * | 2022-04-28 | 2022-08-12 | 河南正通食品科技有限公司 | Preparation method of medium-long chain fatty acid edible oil |
| CN114480518B (en) * | 2022-02-28 | 2024-04-30 | 江南大学 | Method for preparing medium-long carbon chain triglyceride by enzyme method |
-
2019
- 2019-04-17 JP JP2019078911A patent/JP2020174570A/en active Pending
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114480518A (en) * | 2022-02-28 | 2022-05-13 | 江南大学 | Method for preparing medium-long carbon chain triglyceride by enzyme method |
| CN114480518B (en) * | 2022-02-28 | 2024-04-30 | 江南大学 | Method for preparing medium-long carbon chain triglyceride by enzyme method |
| CN114891568A (en) * | 2022-04-28 | 2022-08-12 | 河南正通食品科技有限公司 | Preparation method of medium-long chain fatty acid edible oil |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| DK2006389T3 (en) | Process for preparing concentrated polyunsaturated fatty acid oil | |
| US11781161B2 (en) | Enzymatic enrichment of n-3 fatty acids in the form of glycerides | |
| JP4236128B2 (en) | Essential oil composition | |
| CA2803477C (en) | Process for separating polyunsaturated fatty acids from long chain unsaturated or less saturated fatty acids | |
| WO2008133573A1 (en) | A polyunsaturated fatty acid (pufa) enriched marine oil comprising eicosapentaenoic acid (epa) and docosahexaenoic acid (dha), and a process of production thereof | |
| Wang et al. | Synthesis of docosapentaenoic acid-enriched diacylglycerols by enzymatic glycerolysis of Schizochytrium sp. oil | |
| Diao et al. | Preparation of diacylglycerol from lard by enzymatic glycerolysis and its compositional characteristics | |
| He et al. | Rationale behind the near-ideal catalysis of Candida antarctica lipase A (CAL-A) for highly concentrating ω-3 polyunsaturated fatty acids into monoacylglycerols | |
| Valverde et al. | Concentration of eicosapentaenoic acid (EPA) by selective alcoholysis catalyzed by lipases | |
| Kim et al. | Production of stearidonic acid-rich triacylglycerol via a two-step enzymatic esterification | |
| US20080248187A1 (en) | Mixture containing fatty acid glycerides | |
| JP5753963B1 (en) | Process for producing composition containing lower alcohol fatty acid ester and composition containing lower alcohol fatty acid ester | |
| JP6175198B2 (en) | Method for producing DHA-containing glyceride-containing composition | |
| Li et al. | Preparation of highly pure n-3 PUFA-enriched triacylglycerols by two-step enzymatic reactions combined with molecular Distillation | |
| JP2020174570A (en) | Method for producing polyunsaturated fatty acid and medium chain fatty acid-containing triglyceride | |
| Baik et al. | Enrichment of stearidonic acid from echium oil via a two‐step lipase‐catalyzed esterification | |
| JP6715586B2 (en) | Method for producing highly unsaturated fatty acid | |
| Lee et al. | Preparation of high purity docosahexaenoic acid from microalgae oil in a packed bed reactor via two-step lipase-catalysed esterification | |
| US20120052538A1 (en) | Triglycerides with high content of unsaturated fatty acids | |
| Liu et al. | Production of structured triacylglycerols containing palmitic acids at sn-2 position and docosahexaenoic acids at sn-1, 3 positions | |
| WO2016152174A1 (en) | Method of producing lower alcohol ester of fatty acid -containing composition | |
| JP2018085931A (en) | Method of producing composition containing lower alcohol fatty acid esterified product and composition containing lower alcohol fatty acid esterified product | |
| JP2018082676A (en) | Production method of constituent oils and fats |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| A80 | Written request to apply exceptions to lack of novelty of invention |
Free format text: JAPANESE INTERMEDIATE CODE: A80 Effective date: 20190516 |