JPS6291188A - Production of highly unsaturated fatty acid glyceride - Google Patents
Production of highly unsaturated fatty acid glycerideInfo
- Publication number
- JPS6291188A JPS6291188A JP60232040A JP23204085A JPS6291188A JP S6291188 A JPS6291188 A JP S6291188A JP 60232040 A JP60232040 A JP 60232040A JP 23204085 A JP23204085 A JP 23204085A JP S6291188 A JPS6291188 A JP S6291188A
- Authority
- JP
- Japan
- Prior art keywords
- pufa
- fatty acid
- reaction
- lipase
- 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.)
- Granted
Links
- 235000021122 unsaturated fatty acids Nutrition 0.000 title claims abstract description 10
- 238000004519 manufacturing process Methods 0.000 title claims abstract description 9
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 claims abstract description 30
- 108090001060 Lipase Proteins 0.000 claims abstract description 21
- 239000004367 Lipase Substances 0.000 claims abstract description 21
- 102000004882 Lipase Human genes 0.000 claims abstract description 21
- 235000019421 lipase Nutrition 0.000 claims abstract description 21
- -1 alcohol ester Chemical class 0.000 claims abstract description 11
- 150000004670 unsaturated fatty acids Chemical class 0.000 claims abstract description 6
- 235000011187 glycerol Nutrition 0.000 claims description 12
- 235000014113 dietary fatty acids Nutrition 0.000 claims description 7
- 239000000194 fatty acid Substances 0.000 claims description 7
- 229930195729 fatty acid Natural products 0.000 claims description 7
- 150000004665 fatty acids Chemical class 0.000 claims description 7
- 125000004432 carbon atom Chemical group C* 0.000 claims description 3
- 238000006243 chemical reaction Methods 0.000 abstract description 22
- 125000005456 glyceride group Chemical group 0.000 abstract description 21
- 238000000034 method Methods 0.000 abstract description 15
- 239000002904 solvent Substances 0.000 abstract description 13
- 102000004190 Enzymes Human genes 0.000 abstract description 9
- 108090000790 Enzymes Proteins 0.000 abstract description 9
- 230000000694 effects Effects 0.000 abstract description 6
- 238000005194 fractionation Methods 0.000 abstract description 5
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 abstract description 4
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 abstract description 3
- 241000228143 Penicillium Species 0.000 abstract description 2
- 241000589516 Pseudomonas Species 0.000 abstract description 2
- AHANXAKGNAKFSK-PDBXOOCHSA-N all-cis-icosa-11,14,17-trienoic acid Chemical compound CC\C=C/C\C=C/C\C=C/CCCCCCCCCC(O)=O AHANXAKGNAKFSK-PDBXOOCHSA-N 0.000 abstract description 2
- PRHHYVQTPBEDFE-UHFFFAOYSA-N eicosatrienoic acid Natural products CCCCCC=CCC=CCCCCC=CCCCC(O)=O PRHHYVQTPBEDFE-UHFFFAOYSA-N 0.000 abstract description 2
- 230000003139 buffering effect Effects 0.000 abstract 1
- 229910052799 carbon Inorganic materials 0.000 abstract 1
- 150000001875 compounds Chemical class 0.000 abstract 1
- 235000020777 polyunsaturated fatty acids Nutrition 0.000 description 44
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 11
- 239000000203 mixture Substances 0.000 description 11
- 108010072641 thermostable lipase Proteins 0.000 description 11
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 description 9
- VLKZOEOYAKHREP-UHFFFAOYSA-N n-Hexane Chemical compound CCCCCC VLKZOEOYAKHREP-UHFFFAOYSA-N 0.000 description 7
- 235000020660 omega-3 fatty acid Nutrition 0.000 description 7
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 6
- 239000000047 product Substances 0.000 description 6
- 238000007796 conventional method Methods 0.000 description 5
- 244000005700 microbiome Species 0.000 description 5
- 239000002994 raw material Substances 0.000 description 5
- UFTFJSFQGQCHQW-UHFFFAOYSA-N triformin Chemical compound O=COCC(OC=O)COC=O UFTFJSFQGQCHQW-UHFFFAOYSA-N 0.000 description 5
- 241001465754 Metazoa Species 0.000 description 4
- 150000002148 esters Chemical class 0.000 description 4
- LDVVTQMJQSCDMK-UHFFFAOYSA-N 1,3-dihydroxypropan-2-yl formate Chemical compound OCC(CO)OC=O LDVVTQMJQSCDMK-UHFFFAOYSA-N 0.000 description 3
- ZWEHNKRNPOVVGH-UHFFFAOYSA-N 2-Butanone Chemical compound CCC(C)=O ZWEHNKRNPOVVGH-UHFFFAOYSA-N 0.000 description 3
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 description 3
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 3
- 241000196324 Embryophyta Species 0.000 description 3
- IMNFDUFMRHMDMM-UHFFFAOYSA-N N-Heptane Chemical compound CCCCCCC IMNFDUFMRHMDMM-UHFFFAOYSA-N 0.000 description 3
- 239000002253 acid Substances 0.000 description 3
- 229910001873 dinitrogen Inorganic materials 0.000 description 3
- 125000004494 ethyl ester group Chemical group 0.000 description 3
- 239000003925 fat Substances 0.000 description 3
- 235000019197 fats Nutrition 0.000 description 3
- 238000006317 isomerization reaction Methods 0.000 description 3
- 239000007788 liquid Substances 0.000 description 3
- 239000003921 oil Substances 0.000 description 3
- 235000019198 oils Nutrition 0.000 description 3
- 239000000243 solution Substances 0.000 description 3
- 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 2
- 241000195649 Chlorella <Chlorellales> Species 0.000 description 2
- 102000004157 Hydrolases Human genes 0.000 description 2
- 108090000604 Hydrolases Proteins 0.000 description 2
- 108010093096 Immobilized Enzymes Proteins 0.000 description 2
- KFZMGEQAYNKOFK-UHFFFAOYSA-N Isopropanol Chemical compound CC(C)O KFZMGEQAYNKOFK-UHFFFAOYSA-N 0.000 description 2
- 241000235395 Mucor Species 0.000 description 2
- AMQJEAYHLZJPGS-UHFFFAOYSA-N N-Pentanol Chemical compound CCCCCO AMQJEAYHLZJPGS-UHFFFAOYSA-N 0.000 description 2
- 239000004372 Polyvinyl alcohol Substances 0.000 description 2
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 2
- 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 description 2
- YZXBAPSDXZZRGB-DOFZRALJSA-N arachidonic acid Chemical compound CCCCC\C=C/C\C=C/C\C=C/C\C=C/CCCC(O)=O YZXBAPSDXZZRGB-DOFZRALJSA-N 0.000 description 2
- 238000007664 blowing Methods 0.000 description 2
- 239000000679 carrageenan Substances 0.000 description 2
- 235000010418 carrageenan Nutrition 0.000 description 2
- 229920001525 carrageenan Polymers 0.000 description 2
- 229940113118 carrageenan Drugs 0.000 description 2
- 238000005119 centrifugation Methods 0.000 description 2
- 239000007795 chemical reaction product Substances 0.000 description 2
- 239000000470 constituent Substances 0.000 description 2
- 229940079593 drug Drugs 0.000 description 2
- 239000003814 drug Substances 0.000 description 2
- 230000002255 enzymatic effect Effects 0.000 description 2
- 238000001914 filtration Methods 0.000 description 2
- 235000021323 fish oil Nutrition 0.000 description 2
- 239000011521 glass Substances 0.000 description 2
- 239000003292 glue Substances 0.000 description 2
- ZSIAUFGUXNUGDI-UHFFFAOYSA-N hexan-1-ol Chemical compound CCCCCCO ZSIAUFGUXNUGDI-UHFFFAOYSA-N 0.000 description 2
- 239000003456 ion exchange resin Substances 0.000 description 2
- 229920003303 ion-exchange polymer Polymers 0.000 description 2
- 210000000056 organ Anatomy 0.000 description 2
- 229920002451 polyvinyl alcohol Polymers 0.000 description 2
- 238000010898 silica gel chromatography Methods 0.000 description 2
- 241000894007 species Species 0.000 description 2
- 238000003756 stirring Methods 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 238000004809 thin layer chromatography Methods 0.000 description 2
- UHVMMEOXYDMDKI-JKYCWFKZSA-L zinc;1-(5-cyanopyridin-2-yl)-3-[(1s,2s)-2-(6-fluoro-2-hydroxy-3-propanoylphenyl)cyclopropyl]urea;diacetate Chemical compound [Zn+2].CC([O-])=O.CC([O-])=O.CCC(=O)C1=CC=C(F)C([C@H]2[C@H](C2)NC(=O)NC=2N=CC(=CC=2)C#N)=C1O UHVMMEOXYDMDKI-JKYCWFKZSA-L 0.000 description 2
- WBBQTNCISCKUMU-PDBXOOCHSA-N (13Z,16Z,19Z)-docosatrienoic acid Chemical compound CC\C=C/C\C=C/C\C=C/CCCCCCCCCCCC(O)=O WBBQTNCISCKUMU-PDBXOOCHSA-N 0.000 description 1
- FPRKGXIOSIUDSE-SYACGTDESA-N (2z,4z,6z,8z)-docosa-2,4,6,8-tetraenoic acid Chemical compound CCCCCCCCCCCCC\C=C/C=C\C=C/C=C\C(O)=O FPRKGXIOSIUDSE-SYACGTDESA-N 0.000 description 1
- FHVDTGUDJYJELY-UHFFFAOYSA-N 6-{[2-carboxy-4,5-dihydroxy-6-(phosphanyloxy)oxan-3-yl]oxy}-4,5-dihydroxy-3-phosphanyloxane-2-carboxylic acid Chemical compound O1C(C(O)=O)C(P)C(O)C(O)C1OC1C(C(O)=O)OC(OP)C(O)C1O FHVDTGUDJYJELY-UHFFFAOYSA-N 0.000 description 1
- 241000251468 Actinopterygii Species 0.000 description 1
- 206010003210 Arteriosclerosis Diseases 0.000 description 1
- 241000894006 Bacteria Species 0.000 description 1
- 241001474374 Blennius Species 0.000 description 1
- 241000283690 Bos taurus Species 0.000 description 1
- 241000222120 Candida <Saccharomycetales> Species 0.000 description 1
- 241000238366 Cephalopoda Species 0.000 description 1
- 241000195628 Chlorophyta Species 0.000 description 1
- 241000555825 Clupeidae Species 0.000 description 1
- 241000195493 Cryptophyta Species 0.000 description 1
- 241000065675 Cyclops Species 0.000 description 1
- 241000238557 Decapoda Species 0.000 description 1
- 229920002307 Dextran Polymers 0.000 description 1
- 235000021292 Docosatetraenoic acid Nutrition 0.000 description 1
- 241001373560 Humicola sp. Species 0.000 description 1
- 239000004793 Polystyrene Substances 0.000 description 1
- 101000968491 Pseudomonas sp. (strain 109) Triacylglycerol lipase Proteins 0.000 description 1
- 241000235528 Rhizopus microsporus var. chinensis Species 0.000 description 1
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 1
- 229920002472 Starch Polymers 0.000 description 1
- 241001206171 Stomoxys omega Species 0.000 description 1
- 241000282887 Suidae Species 0.000 description 1
- 238000005377 adsorption chromatography Methods 0.000 description 1
- 230000002411 adverse Effects 0.000 description 1
- 239000003905 agrochemical Substances 0.000 description 1
- 150000001298 alcohols Chemical class 0.000 description 1
- 229940072056 alginate Drugs 0.000 description 1
- 235000010443 alginic acid Nutrition 0.000 description 1
- 229920000615 alginic acid Polymers 0.000 description 1
- 229910052784 alkaline earth metal Inorganic materials 0.000 description 1
- 150000001342 alkaline earth metals Chemical class 0.000 description 1
- PNEYBMLMFCGWSK-UHFFFAOYSA-N aluminium oxide Inorganic materials [O-2].[O-2].[O-2].[Al+3].[Al+3] PNEYBMLMFCGWSK-UHFFFAOYSA-N 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 239000003146 anticoagulant agent Substances 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
- 235000021342 arachidonic acid Nutrition 0.000 description 1
- 229940114079 arachidonic acid Drugs 0.000 description 1
- 208000011775 arteriosclerosis disease Diseases 0.000 description 1
- OGBUMNBNEWYMNJ-UHFFFAOYSA-N batilol Chemical class CCCCCCCCCCCCCCCCCCOCC(O)CO OGBUMNBNEWYMNJ-UHFFFAOYSA-N 0.000 description 1
- 230000004531 blood pressure lowering effect Effects 0.000 description 1
- 239000000872 buffer Substances 0.000 description 1
- 239000007853 buffer solution Substances 0.000 description 1
- AXCZMVOFGPJBDE-UHFFFAOYSA-L calcium dihydroxide Chemical compound [OH-].[OH-].[Ca+2] AXCZMVOFGPJBDE-UHFFFAOYSA-L 0.000 description 1
- 239000000920 calcium hydroxide Substances 0.000 description 1
- 229910001861 calcium hydroxide Inorganic materials 0.000 description 1
- 239000000969 carrier Substances 0.000 description 1
- 239000003054 catalyst Substances 0.000 description 1
- 230000003197 catalytic effect Effects 0.000 description 1
- 239000001913 cellulose Substances 0.000 description 1
- 229920002678 cellulose Polymers 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 238000004587 chromatography analysis Methods 0.000 description 1
- 239000012141 concentrate Substances 0.000 description 1
- 238000001816 cooling Methods 0.000 description 1
- 239000002537 cosmetic Substances 0.000 description 1
- 210000004748 cultured cell Anatomy 0.000 description 1
- 238000000354 decomposition reaction Methods 0.000 description 1
- 238000002845 discoloration Methods 0.000 description 1
- 235000020669 docosahexaenoic acid Nutrition 0.000 description 1
- 229940090949 docosahexaenoic acid Drugs 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 238000006911 enzymatic reaction Methods 0.000 description 1
- 235000004626 essential fatty acids Nutrition 0.000 description 1
- 230000032050 esterification Effects 0.000 description 1
- 238000005886 esterification reaction Methods 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 235000019688 fish Nutrition 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 229930182830 galactose Natural products 0.000 description 1
- 238000004817 gas chromatography Methods 0.000 description 1
- 238000010438 heat treatment Methods 0.000 description 1
- 238000004128 high performance liquid chromatography Methods 0.000 description 1
- 230000007062 hydrolysis Effects 0.000 description 1
- 238000006460 hydrolysis reaction Methods 0.000 description 1
- 239000011261 inert gas Substances 0.000 description 1
- PZLXYMQOCNYUIO-UHFFFAOYSA-N lithium;hydrochloride Chemical compound [Li].Cl PZLXYMQOCNYUIO-UHFFFAOYSA-N 0.000 description 1
- 239000011259 mixed solution Substances 0.000 description 1
- 235000014593 oils and fats Nutrition 0.000 description 1
- 229940012843 omega-3 fatty acid Drugs 0.000 description 1
- 235000020665 omega-6 fatty acid Nutrition 0.000 description 1
- 229940033080 omega-6 fatty acid Drugs 0.000 description 1
- 230000001590 oxidative effect Effects 0.000 description 1
- 239000008363 phosphate buffer Substances 0.000 description 1
- 230000001766 physiological effect Effects 0.000 description 1
- 229920002401 polyacrylamide Polymers 0.000 description 1
- 238000006116 polymerization reaction Methods 0.000 description 1
- 229920002223 polystyrene Polymers 0.000 description 1
- 229910052573 porcelain Inorganic materials 0.000 description 1
- 239000002244 precipitate Substances 0.000 description 1
- 239000000376 reactant Substances 0.000 description 1
- 239000011541 reaction mixture Substances 0.000 description 1
- 230000009257 reactivity Effects 0.000 description 1
- 238000004064 recycling Methods 0.000 description 1
- 239000011347 resin Substances 0.000 description 1
- 229920005989 resin Polymers 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- 238000007127 saponification reaction Methods 0.000 description 1
- 235000019512 sardine Nutrition 0.000 description 1
- 229920006395 saturated elastomer Polymers 0.000 description 1
- 239000013535 sea water Substances 0.000 description 1
- 235000014102 seafood Nutrition 0.000 description 1
- 230000028327 secretion Effects 0.000 description 1
- 235000015170 shellfish Nutrition 0.000 description 1
- 238000007086 side reaction Methods 0.000 description 1
- 230000002269 spontaneous effect Effects 0.000 description 1
- 229910001220 stainless steel Inorganic materials 0.000 description 1
- 239000010935 stainless steel Substances 0.000 description 1
- 239000008107 starch Substances 0.000 description 1
- 235000019698 starch Nutrition 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- RZHACVKGHNMWOP-ZWZRQGCWSA-N tetracosatetraenoic acid n-6 Chemical compound CCCCCCCCCCCCCCC\C=C\C=C\C=C\C=C\C(O)=O RZHACVKGHNMWOP-ZWZRQGCWSA-N 0.000 description 1
- 230000002537 thrombolytic effect Effects 0.000 description 1
- 150000003626 triacylglycerols Chemical class 0.000 description 1
Landscapes
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
Description
【発明の詳細な説明】
fat産業上の利用分野
本発明は、耐熱性リパーゼを用いて高度不飽和脂肪酸グ
リセリドを製造する方法に関するものである。DETAILED DESCRIPTION OF THE INVENTION FIELD OF INDUSTRIAL APPLICATION OF FAT The present invention relates to a method for producing highly unsaturated fatty acid glycerides using a thermostable lipase.
(b)従来の技術
高度不飽和脂肪酸く以下PUFAという)は魚介類、藻
類をはじめとする海産生物、高等動物の臓器、植物、微
生物などに含まれ、その生物種にとって必須脂肪酸であ
ると同時に、近年になり動物や人間の動脈硬化予防に効
果があり、血栓溶解作用、血圧降下作用を有するなどの
生理活性効能が見出され注目をあびている。RE P
U F Aのグリセリドはかかる視点から食品、医薬品
、化粧品。(b) Conventional technology Highly unsaturated fatty acids (hereinafter referred to as PUFA) are found in marine organisms such as fish and shellfish, algae, organs of higher animals, plants, and microorganisms, and are essential fatty acids for the species. In recent years, it has been attracting attention as it has been discovered to be effective in preventing arteriosclerosis in animals and humans, as well as having physiologically active effects such as thrombolytic and blood pressure lowering effects. R.E.P.
From this perspective, UFA glycerides are used in foods, medicines, and cosmetics.
農薬、飼料1診断および分析用試薬などの原料あるいは
基剤として広範囲の分野で有用である。It is useful in a wide range of fields as a raw material or base for agricultural chemicals, feedstuffs, diagnostic and analytical reagents, etc.
該P U F Aのグリセリドはこれまで化学的に合成
、酵素的に変換、あるいは溶剤を用いて調製する方法な
どが試みられてきた。Up to now, attempts have been made to synthesize the glyceride of PUF A chemically, to convert it enzymatically, or to prepare it using a solvent.
すなわち化学的にはグリセリンと該P [J F Aを
酸あるいは塩基触媒共存下に150〜200 ”Cに加
熱し、グリセリドを合成するものであるが、この方法で
はPUFAのトリグリセリドが主生成物になるものの高
温のため該PUFAが重合、異性化、酸化2着色などの
好ましくない副反応を受け、高品質の目的物が得にくい
欠点がある。That is, chemically, glycerin and the P[JFA are heated to 150 to 200''C in the presence of an acid or base catalyst to synthesize glyceride, but in this method, triglyceride of PUFA is the main product. However, due to the high temperature, the PUFA undergoes undesirable side reactions such as polymerization, isomerization, and oxidative discoloration, making it difficult to obtain high-quality target products.
また酵素的に変換する方法としては、いくつかの方法が
試みられているが、例えば特開昭58−165796号
にはリパーゼ(Candida cylindrace
a山来)を用いてもPUFAとグリセリンとのエステル
結合がほとんど加水分解されないことが記載されており
、従ってかかるリパーゼによってはPUFAとグリセリ
ンとの反応もまた起こらないことがわかる。さらに特開
昭59−14793号にもPUFAが従来のリパーゼで
は反応しにくい旨が記載されている。In addition, several methods have been tried for enzymatic conversion, including lipase (Candida cylindrace
It has been described that the ester bond between PUFA and glycerin is hardly hydrolyzed even when using PUFA and glycerin, and therefore it can be seen that the reaction between PUFA and glycerin does not occur depending on such lipase. Furthermore, JP-A-59-14793 also describes that PUFA is difficult to react with conventional lipases.
溶剤を用いる方法としては、特開昭59−71396号
にアセトン、メチルエチルケトン、メタノール。As for a method using a solvent, JP-A-59-71396 discloses acetone, methyl ethyl ketone, and methanol.
エタノールなどの1〜25wt%含水溶剤で魚油を抽出
し、PUFAを含む油脂を濃縮分離する方法が開示され
ているが、かかる溶剤を使用することによるコストアン
プは無視できず、またPUFAO中でも重要なエイコサ
ペンクエン酸(Czo:s・ω−3)含量は30%程度
にとどまっている。A method of extracting fish oil with a 1-25 wt% water-containing solvent such as ethanol and concentrating and separating oils and fats containing PUFA has been disclosed, but the cost increase due to the use of such a solvent cannot be ignored, and it is also important among PUFAO. The eicosapen citric acid (Czo:s.omega.-3) content remains at about 30%.
以上のようにPUFAグリセリドを製造する方法はこれ
までいくつか試みられているが、いずれも満足できるも
のではなかった。As described above, several methods for producing PUFA glycerides have been attempted, but none of them have been satisfactory.
tC1発明が解決しようとする問題点
そこで本発明の目的は、PUFAグリセリドを製造する
に際し、上述したような、化学的合成法における熱履歴
によるPUFAグリセリドの重合。Problems to be Solved by the tC1 Invention Therefore, an object of the present invention is to polymerize PUFA glyceride using thermal history in a chemical synthesis method, as described above, in producing PUFA glyceride.
異性化1着色などの変質、即ら主としてPUFAに起因
する不安定性、酵素的変換法におけるPUFAに対する
従来リパーゼの低ないし非反応性、PUFA トリグリ
セリド含量の低さ、溶剤法における製造コストアンプな
どの従来法の諸問題を解決し、極めて有利な方法で高品
質のPUFAグリセリドを製造せんとするにある。Isomerization 1 Changes such as coloration, i.e. instability mainly due to PUFA, low or non-reactivity of conventional lipases to PUFA in enzymatic conversion methods, low PUFA triglyceride content, production cost increase in solvent methods, etc. The object of the present invention is to solve various problems in the manufacturing process and to produce high quality PUFA glycerides in an extremely advantageous manner.
(d1問題点を解決するための手段
本発明者らは鋭意研究の結果、特定の条件下で反応を行
わせることにより前記の目的が達成できることを見出し
、本発明を完成するに至ったものである。即ち、本発明
はPUFAまたはその低級アルコールエステルとグリセ
リンとを耐熱性リパーゼを用いて反応させることを特徴
とする該PUFAグリセリドの製造法である。(Means for Solving Problem d1) As a result of intensive research, the present inventors discovered that the above object could be achieved by carrying out the reaction under specific conditions, and thus completed the present invention. That is, the present invention is a method for producing PUFA glyceride, which is characterized by reacting PUFA or its lower alcohol ester with glycerin using a heat-stable lipase.
以下に本発明の詳細な説明する。本発明に用いるP[J
FAは魚介類、エビ、イカ5海産クロレラ海苔などの海
産生物、ウシ、ブタなどの高等動物の臓器、植物種子、
カビ、酵母、バクテリアなどの微生物を起源として得る
ことができ、一般に炭素数20以上、不飽和結合を3個
以上有する長鎖長の高度不飽和脂肪酸をいう。これらの
例としては、エイコサトリエン酸(C2゜、3 ω−3
およびω−6)、エイコサテトラエンM (C2゜、4
□ ω−3)、アラキドン酸(C2014,ω−6)、
エイコサペンクエン酸(C2゜83.ω−3)、ドコサ
トリエン酸(Cz□、、ω−6)、 ドコサテトラエン
酸(C2□、4.ω−6)、ドコサヘキサエン酸(C2
□、6.ω−3)、テトラコサトラエン酸(C24:4
゜ω−6)などがあげられ、本発明のPUFAではこれ
らのいわゆる生理活性を有するω−3もしくはω−6脂
肪酸と称されるものを重要な成分とする。なお、本発明
ではこれらのPUFAは単独あるいは混合系で使用する
ことができ、またこれらのPUFAを含む混合脂肪酸と
して使用することもできる。The present invention will be explained in detail below. P[J used in the present invention
FA includes seafood, shrimp, squid, marine organisms such as chlorella seaweed, organs of higher animals such as cows and pigs, plant seeds,
It can be obtained from microorganisms such as mold, yeast, and bacteria, and generally refers to a long chain highly unsaturated fatty acid having 20 or more carbon atoms and 3 or more unsaturated bonds. Examples of these include eicosatrienoic acid (C2°, 3 ω-3
and ω-6), eicosatetraene M (C2°, 4
□ ω-3), arachidonic acid (C2014, ω-6),
Eicosapen citric acid (C2゜83.ω-3), docosatrienoic acid (Cz□,,ω-6), docosatetraenoic acid (C2□, 4.ω-6), docosahexaenoic acid (C2
□, 6. ω-3), tetracosatraenoic acid (C24:4
゜ω-6), etc., and the PUFA of the present invention contains these so-called ω-3 or ω-6 fatty acids having physiological activity as important components. In addition, in the present invention, these PUFAs can be used alone or in a mixed system, and can also be used as a mixed fatty acid containing these PUFAs.
次に本発明でいうPUFAの低級アルコールエステルの
低級アルコールとは炭素数6以下のアルコールをさし、
メタノール、エタノール、プロパツール、イソプロパツ
ール、ブタ5ノール、ペンタノール、ヘキサノールなど
を例示することができる。PUFAとこれらのアルコー
ルのエステルは常法によりエステル化して得ることがで
き、これらの単独あるいは混合物として使用することが
でできる。Next, the lower alcohol of the lower alcohol ester of PUFA in the present invention refers to an alcohol having 6 or less carbon atoms,
Examples include methanol, ethanol, propatool, isopropanol, buta5ol, pentanol, hexanol, and the like. Esters of PUFA and these alcohols can be obtained by esterification by a conventional method, and can be used alone or as a mixture.
従来の油脂加水分解酵素は一般に30〜45℃で使用さ
れ、中性付近1常圧の温和な反応条件で触媒作用を示す
が、上述のようなPUFAまたはそのエステルに作用す
る酵素は見出されていない。Conventional oil and fat hydrolases are generally used at 30 to 45°C and exhibit catalytic activity under mild reaction conditions near neutrality and normal pressure, but enzymes that act on PUFAs or their esters as described above have not been found. Not yet.
このためPUFAグリセリドを酵素的に製造する効果的
な方法も確立されていない。しかしながら、本発明者ら
は、耐熱性リパーゼを適用することによってPUFAグ
リセリドを製造することを可能ならしめることができた
。該耐熱性リパーゼは50°C以上でも活性を維持でき
るものであれば良く、このような酵素は動植物あるいは
微生物の生体ないし、分泌液から単離、精製して採取す
ることができるが、微生物由来のものが簡便である。こ
れさの例としては、シュードモナス属のPseudom
onasfluorescens 由来のリパーゼ、ペ
ニシリウム屈のPenicillium cyclop
ium由来のリパーゼ、アルカリ土類金属の微工研菌寄
第3783号株由来のリパーゼ、ヒューミコラ属のHu
micola lanuginose由来のリパーゼ、
ムコール属のl’lu(ormiehei由来のリパー
ゼ、リゾプス属のRh1zopus chinensi
s由来のリパーゼなどをあげることができる。なお、本
発明はこれらの種の微生物に限定されるものではなく、
これらの種に属する野生株、変異株、栄養要求性株、薬
剤耐性株からの耐熱性リパーゼを用いてもさしつかえな
い。また、該耐熱性リパーゼはこれを固定化物としたも
のでも使用できる。例えばセルロース、デキストラン、
ポリスチレン、ポリアクリルアミド、ポリビニルアルコ
ール、イオン交換樹脂、磁製体、活性炭、セライト、ア
ルミナ、光架橋性あるいは光硬化性樹脂、アルギン酸塩
、カラキーナンなどの酵素固定化用担体に吸着。For this reason, an effective method for enzymatically producing PUFA glycerides has not been established. However, the inventors were able to make it possible to produce PUFA glycerides by applying thermostable lipases. The thermostable lipase may be one that can maintain its activity even at temperatures above 50°C, and such enzymes can be isolated and purified from living organisms or secretions of animals, plants, or microorganisms; The one is simple. An example of this is Pseudomonas spp.
Lipase from onasfluorescens, Penicillium cyclop
Lipase derived from alkaline earth metal microorganism strain No. 3783, Hu of Humicola sp.
lipase from micola lanuginose,
Lipase from Mucor genus l'lu (ormiehei, Rhizopus chinensi
Examples include lipase derived from S. Note that the present invention is not limited to these types of microorganisms,
Thermostable lipases from wild strains, mutant strains, auxotrophic strains, and drug-resistant strains belonging to these species may be used. Furthermore, the thermostable lipase can also be used as an immobilized product. For example, cellulose, dextran,
Adsorbs to enzyme immobilization carriers such as polystyrene, polyacrylamide, polyvinyl alcohol, ion exchange resin, porcelain, activated carbon, celite, alumina, photocrosslinkable or photocurable resin, alginate, carakeenan, etc.
イオン結合、共有結合あるいは包括させた耐熱性リパー
ゼを使用することができる。Ionic, covalent or entrapped thermostable lipases can be used.
本発明の方法により、P [J l;’ Aまたはその
低級アルコールエステルとグリセリンとを耐熱性リパー
ゼを用いて反応させ、該PUFAグリセリドを製造する
には次のようにする。即ち、反応原料であるPUFAま
たはその低級アルコールエステルおよびグリセリンをガ
ラス製あるいはステンレス製容器に採り、必要に応じて
不゛活性有殿溶媒例えばn−ヘキサン、n−へブタンな
どを加え、さらに連星の水、緩衝液を適宜添加し、耐熱
性リパーゼもしくはその固定化物を加えたのら、攪拌も
しくは振とうしながら不活性気体例えば窒素ガスなどを
吹き込みつつ、p H3〜12、好ましくはpH5〜I
Oの下、50℃以上好ましくは50〜70℃に昇温し、
5〜120時間反応させる。ここで゛本発明で用いる酵
素は50°C未満でも反応は進行するが、長時間を要す
ること、また70℃を超える温度では耐熱性リパーゼと
いえども安定性が小さくなり、酵素の失活を招くこと及
び高温による不飽和基の劣化のおそれなどから、50〜
70°Cで反応を行うのが望ましい。反応の進行状況は
、TLC(薄層クロマトグラフィー)、Ct、C(ガス
クロマトグラフィー)あるいはI(PLC(高速?rf
t体クロマトグラフィー)などでPUFAまたはそのエ
ステル含量の減少、該PUFAグリセリド中のモノグリ
セリド、ジグリセリドおよびトリグリセリド含量の増加
を測定することによりチェ”/りできる。反応終了後、
反応物を80〜90℃に短時間加熱して酵素を失活させ
、濾過もしくは遠心分離により酵素を除き、さらに溶剤
分別、吸着クロマトグラフィーなどの分画手法を用いて
未反応物を除去し、目的のPUFAグリセリドを製造す
ることができる。なお、本発明では固定化酵素を円筒容
器に詰め、反応液を一方から流し込み、カラム方式のり
アクタ−として反応させることもでき、あるいは反応液
中に固定化酵素を浮遊させる分散方式のりアクタ−とし
ても反応を行うことができる。According to the method of the present invention, P [J l;' A or its lower alcohol ester and glycerin are reacted using a thermostable lipase to produce the PUFA glyceride as follows. That is, the reaction raw materials, PUFA or its lower alcohol ester, and glycerin are placed in a glass or stainless steel container, and if necessary, an inactive precipitate solvent such as n-hexane or n-hebutane is added, and then a binary starch is added. After adding water and a buffer solution as appropriate, and adding heat-stable lipase or its immobilized product, the pH is adjusted to pH 3 to 12, preferably pH 5 to I, while stirring or shaking and blowing an inert gas such as nitrogen gas.
The temperature is raised to 50°C or higher, preferably 50 to 70°C under O,
React for 5 to 120 hours. Here, although the enzyme used in the present invention can proceed at temperatures below 50°C, it takes a long time, and at temperatures above 70°C, even heat-stable lipases become less stable, and the enzyme may be deactivated. 50 ~
It is desirable to carry out the reaction at 70°C. The progress of the reaction can be monitored by TLC (thin layer chromatography), Ct, C (gas chromatography) or I (PLC (high speed? rf
This can be checked by measuring the decrease in the content of PUFA or its ester and the increase in the monoglyceride, diglyceride, and triglyceride content in the PUFA glyceride using t-isomer chromatography. After completion of the reaction,
The reaction product is heated to 80 to 90°C for a short time to inactivate the enzyme, the enzyme is removed by filtration or centrifugation, and unreacted products are removed using a fractionation method such as solvent fractionation or adsorption chromatography. A desired PUFA glyceride can be produced. In addition, in the present invention, it is also possible to pack the immobilized enzyme in a cylindrical container and pour the reaction solution from one side to cause the reaction to occur as a column-type glue actor, or as a dispersion-type glue actor in which the immobilized enzyme is suspended in the reaction solution. can also carry out the reaction.
(el実施例
実施例1
緑藻類の一種である海産クロレラを海水中で培養し、濃
縮した培養細胞を破砕、溶剤抽出および溶剤分画してガ
ラクトースおよびエイコサベンクエン酸(CZ。、5.
ω−3)を主要構成成分とするυ月脂質を得た。これを
常法によりケン化分解し、溶剤抽出してPUFAを得た
。脂肪酸組成:C2゜86.ω−3(77%)、C2゜
、4.ω−6(8%)。(EL Examples Example 1 Marine chlorella, a type of green algae, is cultured in seawater, and the concentrated cultured cells are crushed, solvent extracted, and solvent fractionated to produce galactose and eicosaben citric acid (CZ).
A υ-moon lipid containing ω-3) as a main component was obtained. This was saponified and decomposed by a conventional method and extracted with a solvent to obtain PUFA. Fatty acid composition: C2°86. ω-3 (77%), C2°, 4. ω-6 (8%).
C18,2(3%) 、Cls:+ (3%)、Cl6
F。(5%)であった。C18,2 (3%), Cls:+ (3%), Cl6
F. (5%).
このPUFA75 gおよびグリセリン10g。75 g of this PUFA and 10 g of glycerin.
n−ヘキサン100mffを冷却前付500mff三ツ
ロフラスコに採り、0.3Mリン酸緩衝液(pl(7,
0)0.8mffおよび耐熱性リパーゼ(Ilumic
olaIanug i nosa山来、由来製薬(+@
製、商品名rLipaseCEJ)0.1gを添加し、
窒素ガスを吹き込みながら60℃で72時間攪拌(20
Orpm)した。Transfer 100 mff of n-hexane to a 500 mff Mitsuro flask with a cooling prefix, add 0.3 M phosphate buffer (pl (7,
0) 0.8mff and thermostable lipase (Ilumic
olaIanug i nosa Yamaki, origin pharmaceutical (+@
0.1 g of Lipase CEJ, manufactured by Co., Ltd., trade name:
Stir at 60°C for 72 hours while blowing nitrogen gas (20
Orpm).
反応終了後、85“Cに5分間加熱し、遠心分離操作で
酵素を除き、シリカゲルカラムクロマトグラフィーで未
反応のPUFAを除去した後、HP LC分析により反
応物のグリセリド組成を求めたところ、トリグリセリド
(86%)、ジグリセリド(1)%)、モノグリセリド
(3%)であった。After the reaction was completed, the mixture was heated to 85"C for 5 minutes, the enzyme was removed by centrifugation, and unreacted PUFA was removed by silica gel column chromatography. The glyceride composition of the reaction product was determined by HPLC analysis. (86%), diglyceride (1%), and monoglyceride (3%).
また、グリセリド成分の構成脂肪酸をGLCにより分析
した結果、原料であるPUFAとほぼ同組成であった。Furthermore, as a result of analyzing the constituent fatty acids of the glyceride component by GLC, it was found that the composition was almost the same as that of the raw material PUFA.
実施例2
実施例1で得たPUFAを常法によりエタノールでPU
FAエチルエステルに変換した。該PUFAエチルエス
テル50g、グリセリン6.7gを200mA三角フラ
スコに採り、0.5 M l−リス塩H1)街液(p
II 8.5) 0.5mlおよび固定化耐熱性リパー
ゼ(Mucor m1ehei由来のリパーゼを弱塩基
性イオン交換樹脂に固定化したもの、デンマーク・ノボ
インダストリー社製、商品名rLipase3AJ)0
.2gを添加し、窒素ガス気流中、65℃で90時間振
とう(200rpm)した。反応終了後、80°Cに5
分間加熱し固定化物を濾過で除き、シリカゲルカラムク
ロマトグラフィーで未反応物(P U F A)を除去
した。生成物の組成ニトリグリセリド(25%)、ジグ
リセリド(58%)、モノグリセリド(17%)であり
、その脂肪酸組成は原料とほぼ同じであった。Example 2 PUFA obtained in Example 1 was purified with ethanol using a conventional method.
Converted to FA ethyl ester. 50 g of the PUFA ethyl ester and 6.7 g of glycerin were placed in a 200 mA Erlenmeyer flask, and 0.5 M l-lith salt H1) street liquor (p
II 8.5) 0.5 ml and immobilized thermostable lipase (lipase derived from Mucor m1ehei immobilized on weakly basic ion exchange resin, manufactured by Novo Industries, Denmark, trade name rLipase3AJ) 0
.. 2 g was added thereto, and the mixture was shaken (200 rpm) at 65° C. for 90 hours in a nitrogen gas stream. After the reaction is complete, heat to 80°C for 5 minutes.
After heating for a minute, the immobilized substances were removed by filtration, and unreacted substances (PUFA) were removed by silica gel column chromatography. The composition of the product was nitriglyceride (25%), diglyceride (58%), and monoglyceride (17%), and its fatty acid composition was almost the same as that of the raw material.
実施例3
実施例2で使用した固定化耐熱性リパーゼ(ノボインダ
ストリー社製、商品名「Lipage 3 A J )
をガラス管(2cm径X 3 、Ocm長)シこ詰め、
実施例2記載のPUFAエチルエステル50g、グリセ
リン5g、n−ヘプタン50mA、0.3M!−リス塩
酸緩衝液(pH7,5)0.8m/およびポリビニルア
ルコール(n=1750)0.05gを別容器中でホモ
ミキサーで2000r p m、 15分間処理した
混合液となし、この混合液を液送ポンプを介して上記カ
ラムの下部ア・ら10m1!/時間の流速で送入させ、
上部から流出する液を上記の別容器中に戻すような液循
環系をつくり、60℃で80時間リサイクルを行った。Example 3 Immobilized thermostable lipase used in Example 2 (manufactured by Novo Industries, trade name “Lipage 3 A J”)
Filled with glass tube (2cm diameter x 3, Ocm length),
50 g of PUFA ethyl ester described in Example 2, 5 g of glycerin, 50 mA of n-heptane, 0.3 M! - 0.8 m of lithium-hydrochloric acid buffer (pH 7,5) and 0.05 g of polyvinyl alcohol (n = 1750) were treated in a separate container with a homomixer at 2000 rpm for 15 minutes to form a mixed solution. 10m1 of the lower part of the above column via the liquid feed pump! / hour flow rate,
A liquid circulation system was created to return the liquid flowing out from the top into the separate container mentioned above, and recycling was performed at 60°C for 80 hours.
反応液の一部を抜き取りその組成をTLCで分析したと
ころ、主成分はジグリセリドであった。When a portion of the reaction solution was extracted and its composition was analyzed by TLC, the main component was diglyceride.
実施例4
いわし由来の魚油を常法により非耐熱性リパーゼ(名糖
産業側製、[リパーゼM Y J )を用いて30°c
、 6時間反応させ加水分解した。反応物に水酸化カ
ルシウム飽和水溶液を加え、遠心分離して油層を回収し
た後、常法によりケン化分解、ヘキサン抽出してPUF
Aの濃縮物を得た。GLC分析による主要脂肪酸組成は
C2゜86.n=3(38%)、C2□、6.n=3
(25%)、C2□81.n=3 (5%)、C2゜、
l(5%)、C2□、I(3%)。Example 4 Fish oil derived from sardines was heated at 30°C using a non-heat-stable lipase (manufactured by Meito Sangyo Co., Ltd., [Lipase M Y J]) using a conventional method.
, and was allowed to react for 6 hours for hydrolysis. A saturated aqueous solution of calcium hydroxide is added to the reaction mixture, centrifuged to collect an oil layer, followed by saponification and decomposition using a conventional method and extraction with hexane to obtain PUF.
A concentrate of A was obtained. The main fatty acid composition according to GLC analysis is C2°86. n=3 (38%), C2□, 6. n=3
(25%), C2□81. n=3 (5%), C2°,
l (5%), C2□, I (3%).
C2゜、4.n=6 (2%)、その他(22%)であ
った。このpuFA?fi縮物100 g、グリセリン
10g、水0.5 m II 、耐熱性リパーゼ(Ps
eudomonasf 1uorescens由来、天
野製薬0@製、商品名rLIpasePj)0.15g
、セライト012gを用い、実施例1と同様に反応させ
、反応物の組成を求めたところ、トリグリセリド(80
%)、ジグリセリド(13%)、モノグリセリド(7%
)であり、それらの構成脂肪酸は原料のPUFA?Q縮
物とほぼ同じであった。C2°, 4. n=6 (2%), and others (22%). This puFA? 100 g of ficondensate, 10 g of glycerin, 0.5 m of water, thermostable lipase (Ps
derived from eudomonasf 1uorescens, manufactured by Amano Pharmaceutical 0@, trade name rLIpasePj) 0.15g
, Celite 012g was reacted in the same manner as in Example 1, and the composition of the reactant was determined. Triglyceride (80
%), diglycerides (13%), monoglycerides (7%)
), and their constituent fatty acids are the raw material PUFA? It was almost the same as the Q condensate.
(f)発明の効果 本発明の効果は次の通りである。(f) Effect of invention The effects of the present invention are as follows.
■ 従来、30〜45°Cで使用していた通常の油脂加
水分解酵素では反応が不可能とされていたPUFAまた
はそのエステルを、耐熱性リパーゼを用いることにより
反応させることが可能となり、咳P U F Aのグリ
セリドを製造することができるようになった。■ By using heat-stable lipase, it is now possible to react PUFAs or their esters, which were previously thought to be impossible to react with normal fat and oil hydrolases used at 30-45°C. It is now possible to produce UFA glyceride.
■ かかる反応は50〜70℃という比較的低温のため
、熱安定製の悪いPUFAに対し、重合。(2) Since this reaction is performed at a relatively low temperature of 50 to 70°C, PUFA, which has poor thermal stability, is polymerized.
異性化などの悪影響を与えず、高品質のPUFAグリセ
リドが製造可能となった。High-quality PUFA glycerides can now be produced without adverse effects such as isomerization.
■ 従来の酵素法により得られるPUFAグリセリドは
トリグリセリド含量を増大させることに限界があったが
、本発明の方法ではほぼ完全なPUFAトリグリセリド
を製造できる。(2) PUFA glycerides obtained by conventional enzymatic methods had a limit in increasing the triglyceride content, but the method of the present invention can produce almost complete PUFA triglycerides.
■ 自然分別あるいは溶剤分別法に比べ、常温に近い温
度で反応でき、また多量の溶剤を必要とせず、PUFA
グリセリドの製造におけるコスト・ダウンが期待できる
。■ Compared to natural fractionation or solvent fractionation methods, the reaction can be carried out at temperatures close to room temperature, and large amounts of solvent are not required.
Cost reduction in glyceride production can be expected.
特許出願人 日清製油株式会社
手続補正書(自発)
昭和ρ年〃月6日
特許庁長官 宇 賀 道6部゛′、(゛漬1、事件の
表示
昭和60年特許願第232040号
2、発明の名称
高度不飽和脂肪酸グリセリドの製造法
3、補正をする者
事件との関係 特許出願人
郵便番号 221
住 所 神奈川県横浜市神奈用区千若町1−3名
称 日清製油株式会社 研究断電 話 0
45 (461)0181−一゛1)
7、゛・・1(゛・・8
fと・′、・
4゜補正の対象
(1)明細書の発明の詳細な説明の欄
5、補正の内容
(1) 明細書第5−頁1)行「テトラコサトラエン
酸」を「テトラコサテトラエン酸」と訂正する。Patent Applicant: Nisshin Oil Co., Ltd. Procedural Amendment (Spontaneous) Date: June 6, 1985 Director General of the Patent Office Uga Michi 6th Department ``(゛zuke 1, Case Indication 1985 Patent Application No. 232040 2, Title of the invention Process for producing highly unsaturated fatty acid glycerides 3, Relationship to the amended case Patent applicant Postal code 221 Address 1-3 Chiwaka-cho, Kanayō-ku, Yokohama-shi, Kanagawa Prefecture
Name Nisshin Oil Co., Ltd. Research call outage Call 0
45 (461)0181-1゛1) 7,゛...1(゛...8 f and...',・4゜Object of amendment (1) Detailed explanation of the invention in the specification column 5, Contents of amendment (1) On page 5-1 of the specification, line "Tetracosatraenoic acid" is corrected to "Tetracosatetraenoic acid."
(2)明細書第6下から1行〜第7頁1行「これさの例
としては、」を「これらの例としては、」と訂正する。(2) "As an example of this," is corrected from line 1 from the bottom of page 6 to page 7 of the specification as "these examples are."
(3)同第7頁5行r 1anuginoseJをr
lanuginosa」と訂正する。(3) Same page 7 line 5 r 1anuginoseJ
lanuginosa”.
(4) 同第7頁下から3行「カラギーナンなどの」
を「カラギーナンなどの」と訂正する。(4) Page 7, 3 lines from the bottom: “Carrageenan, etc.”
should be corrected as "carrageenan, etc."
(5)明細書第13頁1)行〜12行rrLIpase
P」をr r Lipase P Jと訂正する。(5) Specification page 13, lines 1) to 12 rrLIpase
Correct "P" to r r Lipase P J.
(6)明細書第14頁7行「熱安定型の」を「熱安定性
の」と訂正する。(6) On page 14 of the specification, line 7, "thermally stable" is corrected to "thermally stable."
Claims (2)
テルとグリセリンとを耐熱性リパーゼを用いて反応させ
ることを特徴とする高度不飽和脂肪酸グリセリドの製造
法。(1) A method for producing a highly unsaturated fatty acid glyceride, which comprises reacting a highly unsaturated fatty acid or its lower alcohol ester with glycerin using a heat-stable lipase.
合を3個以上有する脂肪酸である特許請求の範囲第(1
)項記載の製造法。(2) The highly unsaturated fatty acid is a fatty acid having 20 or more carbon atoms and 3 or more unsaturated bonds.
) The manufacturing method described in section 2.
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP60232040A JPS6291188A (en) | 1985-10-17 | 1985-10-17 | Production of highly unsaturated fatty acid glyceride |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP60232040A JPS6291188A (en) | 1985-10-17 | 1985-10-17 | Production of highly unsaturated fatty acid glyceride |
Publications (2)
Publication Number | Publication Date |
---|---|
JPS6291188A true JPS6291188A (en) | 1987-04-25 |
JPH0582197B2 JPH0582197B2 (en) | 1993-11-17 |
Family
ID=16933019
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP60232040A Granted JPS6291188A (en) | 1985-10-17 | 1985-10-17 | Production of highly unsaturated fatty acid glyceride |
Country Status (1)
Country | Link |
---|---|
JP (1) | JPS6291188A (en) |
Cited By (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO1991016443A1 (en) * | 1990-04-18 | 1991-10-31 | Novo Nordisk A/S | Process for preparation of triglyceride and triglyceride composition |
JPH0595792A (en) * | 1991-10-03 | 1993-04-20 | Agency Of Ind Science & Technol | Production of oil and fat containing concentrated highly unsaturated fatty acid |
US5945318A (en) * | 1994-03-08 | 1999-08-31 | Norsk Hydro A.S. | Refining oil compositions |
JP2010183881A (en) * | 2009-02-13 | 2010-08-26 | Kao Corp | Method for producing oil and fat having high docosahexaenoic acid content |
EP2602308A2 (en) | 2002-11-14 | 2013-06-12 | Pronova BioPharma Norge AS | Lipase-catalysed esterification of marine oil |
Citations (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JPS60234588A (en) * | 1984-05-07 | 1985-11-21 | Asahi Denka Kogyo Kk | Production of long-chain highly unsaturated fatty acid alcohol ester |
-
1985
- 1985-10-17 JP JP60232040A patent/JPS6291188A/en active Granted
Patent Citations (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JPS60234588A (en) * | 1984-05-07 | 1985-11-21 | Asahi Denka Kogyo Kk | Production of long-chain highly unsaturated fatty acid alcohol ester |
Cited By (6)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO1991016443A1 (en) * | 1990-04-18 | 1991-10-31 | Novo Nordisk A/S | Process for preparation of triglyceride and triglyceride composition |
US5604119A (en) * | 1990-04-18 | 1997-02-18 | Novo Nordisk A/S | Process for producing triglycerides from glycerol and long-chain polyunsaturated fatty acids using lipase from candida antarctica |
JPH0595792A (en) * | 1991-10-03 | 1993-04-20 | Agency Of Ind Science & Technol | Production of oil and fat containing concentrated highly unsaturated fatty acid |
US5945318A (en) * | 1994-03-08 | 1999-08-31 | Norsk Hydro A.S. | Refining oil compositions |
EP2602308A2 (en) | 2002-11-14 | 2013-06-12 | Pronova BioPharma Norge AS | Lipase-catalysed esterification of marine oil |
JP2010183881A (en) * | 2009-02-13 | 2010-08-26 | Kao Corp | Method for producing oil and fat having high docosahexaenoic acid content |
Also Published As
Publication number | Publication date |
---|---|
JPH0582197B2 (en) | 1993-11-17 |
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