JPH0286797A - Production of optically active ester of 2-alkanol - Google Patents
Production of optically active ester of 2-alkanolInfo
- Publication number
- JPH0286797A JPH0286797A JP23937388A JP23937388A JPH0286797A JP H0286797 A JPH0286797 A JP H0286797A JP 23937388 A JP23937388 A JP 23937388A JP 23937388 A JP23937388 A JP 23937388A JP H0286797 A JPH0286797 A JP H0286797A
- Authority
- JP
- Japan
- Prior art keywords
- reaction
- alkanol
- optically active
- ester
- lipase
- 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
- 150000002148 esters Chemical class 0.000 title claims abstract description 27
- 238000004519 manufacturing process Methods 0.000 title claims description 8
- 102000004882 Lipase Human genes 0.000 claims abstract description 23
- 108090001060 Lipase Proteins 0.000 claims abstract description 23
- 239000004367 Lipase Substances 0.000 claims abstract description 23
- 235000019421 lipase Nutrition 0.000 claims abstract description 23
- UFTFJSFQGQCHQW-UHFFFAOYSA-N triformin Chemical compound O=COCC(OC=O)COC=O UFTFJSFQGQCHQW-UHFFFAOYSA-N 0.000 claims abstract description 9
- 238000006243 chemical reaction Methods 0.000 abstract description 48
- UYXTWWCETRIEDR-UHFFFAOYSA-N Tributyrin Chemical compound CCCC(=O)OCC(OC(=O)CCC)COC(=O)CCC UYXTWWCETRIEDR-UHFFFAOYSA-N 0.000 abstract description 23
- VLKZOEOYAKHREP-UHFFFAOYSA-N n-Hexane Chemical compound CCCCCC VLKZOEOYAKHREP-UHFFFAOYSA-N 0.000 abstract description 12
- 239000002904 solvent Substances 0.000 abstract description 4
- 238000003756 stirring Methods 0.000 abstract description 4
- 230000002829 reductive effect Effects 0.000 abstract description 3
- 239000003814 drug Substances 0.000 abstract description 2
- 239000007788 liquid Substances 0.000 abstract description 2
- 239000004973 liquid crystal related substance Substances 0.000 abstract description 2
- 239000000463 material Substances 0.000 abstract description 2
- 102000004190 Enzymes Human genes 0.000 description 26
- 108090000790 Enzymes Proteins 0.000 description 26
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 17
- 230000000694 effects Effects 0.000 description 14
- 238000000034 method Methods 0.000 description 13
- 238000005809 transesterification reaction Methods 0.000 description 10
- 235000014113 dietary fatty acids Nutrition 0.000 description 9
- 239000000194 fatty acid Substances 0.000 description 9
- 229930195729 fatty acid Natural products 0.000 description 9
- 150000004665 fatty acids Chemical class 0.000 description 8
- 230000003287 optical effect Effects 0.000 description 7
- 239000000243 solution Substances 0.000 description 7
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 6
- 125000000217 alkyl group Chemical group 0.000 description 6
- 229910052799 carbon Inorganic materials 0.000 description 6
- 239000011148 porous material Substances 0.000 description 5
- 125000004432 carbon atom Chemical group C* 0.000 description 4
- 239000003960 organic solvent Substances 0.000 description 4
- UHOVQNZJYSORNB-UHFFFAOYSA-N Benzene Chemical compound C1=CC=CC=C1 UHOVQNZJYSORNB-UHFFFAOYSA-N 0.000 description 3
- 241000589516 Pseudomonas Species 0.000 description 3
- 230000007062 hydrolysis Effects 0.000 description 3
- 238000006460 hydrolysis reaction Methods 0.000 description 3
- 230000002401 inhibitory effect Effects 0.000 description 3
- 230000035484 reaction time Effects 0.000 description 3
- 230000002194 synthesizing effect Effects 0.000 description 3
- KBPLFHHGFOOTCA-UHFFFAOYSA-N 1-Octanol Chemical compound CCCCCCCCO KBPLFHHGFOOTCA-UHFFFAOYSA-N 0.000 description 2
- WJZIPMQUKSTHLV-UHFFFAOYSA-N 2-ethyldecanoic acid Chemical compound CCCCCCCCC(CC)C(O)=O WJZIPMQUKSTHLV-UHFFFAOYSA-N 0.000 description 2
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 description 2
- IMNFDUFMRHMDMM-UHFFFAOYSA-N N-Heptane Chemical compound CCCCCCC IMNFDUFMRHMDMM-UHFFFAOYSA-N 0.000 description 2
- 239000002253 acid Substances 0.000 description 2
- 239000003125 aqueous solvent Substances 0.000 description 2
- 230000015572 biosynthetic process Effects 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 238000001914 filtration Methods 0.000 description 2
- 230000003301 hydrolyzing effect Effects 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- SJWFXCIHNDVPSH-UHFFFAOYSA-N octan-2-ol Chemical compound CCCCCCC(C)O SJWFXCIHNDVPSH-UHFFFAOYSA-N 0.000 description 2
- 210000000496 pancreas Anatomy 0.000 description 2
- 239000000843 powder Substances 0.000 description 2
- 239000000047 product Substances 0.000 description 2
- 239000002994 raw material Substances 0.000 description 2
- 230000002441 reversible effect Effects 0.000 description 2
- 239000007787 solid Substances 0.000 description 2
- 239000000758 substrate Substances 0.000 description 2
- 238000003786 synthesis reaction Methods 0.000 description 2
- VZGDMQKNWNREIO-UHFFFAOYSA-N tetrachloromethane Chemical compound ClC(Cl)(Cl)Cl VZGDMQKNWNREIO-UHFFFAOYSA-N 0.000 description 2
- VMPHSYLJUKZBJJ-UHFFFAOYSA-N trilaurin Chemical compound CCCCCCCCCCCC(=O)OCC(OC(=O)CCCCCCCCCCC)COC(=O)CCCCCCCCCCC VMPHSYLJUKZBJJ-UHFFFAOYSA-N 0.000 description 2
- SYTBZMRGLBWNTM-SNVBAGLBSA-N (R)-flurbiprofen Chemical compound FC1=CC([C@H](C(O)=O)C)=CC=C1C1=CC=CC=C1 SYTBZMRGLBWNTM-SNVBAGLBSA-N 0.000 description 1
- DURPTKYDGMDSBL-UHFFFAOYSA-N 1-butoxybutane Chemical compound CCCCOCCCC DURPTKYDGMDSBL-UHFFFAOYSA-N 0.000 description 1
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical group [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 1
- ZAFNJMIOTHYJRJ-UHFFFAOYSA-N Diisopropyl ether Chemical compound CC(C)OC(C)C ZAFNJMIOTHYJRJ-UHFFFAOYSA-N 0.000 description 1
- NHTMVDHEPJAVLT-UHFFFAOYSA-N Isooctane Chemical compound CC(C)CC(C)(C)C NHTMVDHEPJAVLT-UHFFFAOYSA-N 0.000 description 1
- 101710084373 Lipase 1 Proteins 0.000 description 1
- 101710084376 Lipase 3 Proteins 0.000 description 1
- 241000228143 Penicillium Species 0.000 description 1
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 1
- PMZURENOXWZQFD-UHFFFAOYSA-L Sodium Sulfate Chemical compound [Na+].[Na+].[O-]S([O-])(=O)=O PMZURENOXWZQFD-UHFFFAOYSA-L 0.000 description 1
- 241000179532 [Candida] cylindracea Species 0.000 description 1
- 230000002776 aggregation Effects 0.000 description 1
- 238000004220 aggregation Methods 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 239000000872 buffer Substances 0.000 description 1
- 239000007853 buffer solution Substances 0.000 description 1
- 238000004440 column chromatography Methods 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 239000012043 crude product Substances 0.000 description 1
- 230000009849 deactivation Effects 0.000 description 1
- 230000007423 decrease Effects 0.000 description 1
- 230000003111 delayed effect Effects 0.000 description 1
- JVSWJIKNEAIKJW-UHFFFAOYSA-N dimethyl-hexane Natural products CCCCCC(C)C JVSWJIKNEAIKJW-UHFFFAOYSA-N 0.000 description 1
- 230000032050 esterification Effects 0.000 description 1
- 238000005886 esterification reaction Methods 0.000 description 1
- 150000002168 ethanoic acid esters Chemical class 0.000 description 1
- -1 fatty acid esters Chemical class 0.000 description 1
- 238000004817 gas chromatography Methods 0.000 description 1
- 230000003100 immobilizing effect Effects 0.000 description 1
- TVMXDCGIABBOFY-UHFFFAOYSA-N octane Chemical compound CCCCCCCC TVMXDCGIABBOFY-UHFFFAOYSA-N 0.000 description 1
- 239000004006 olive oil Substances 0.000 description 1
- 235000008390 olive oil Nutrition 0.000 description 1
- 230000004845 protein aggregation Effects 0.000 description 1
- 239000011541 reaction mixture Substances 0.000 description 1
- 230000009257 reactivity Effects 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 238000007086 side reaction Methods 0.000 description 1
- 239000000741 silica gel Substances 0.000 description 1
- 229910002027 silica gel Inorganic materials 0.000 description 1
- 150000003626 triacylglycerols Chemical class 0.000 description 1
- 238000005292 vacuum distillation Methods 0.000 description 1
Abstract
Description
【発明の詳細な説明】
（産業上の利用分野）
本発明は酵素を用いる２−アルカノールの光学活性エス
テルの製造方法に関する。DETAILED DESCRIPTION OF THE INVENTION (Industrial Field of Application) The present invention relates to a method for producing optically active esters of 2-alkanols using enzymes.
（従来の技術）
従来、酵素を用いた２−アルカノールの光学分割や２−
アルカノールの光学活性エステルの合成方法としては、
下記式のように２−アルカノールの脂肪酸エステルを特
異的に加水分解する方法が（式中、Ｒ１１は炭素数ｌ〜
１２のアルキル基と、Ｒ１２は炭素数１−１０のアルキ
ル基を示す。）しかし、この種の反応を触媒する酵素は
反応によって生成する脂肪酸による阻害効果を受は易く
、反応が途中て停止したり１反応が遅くなるという欠点
を持っている。また、下記式のように加水分解の逆反応
であるエステル化を用いて光学活性エステルを合成する
方法も知られている。(Prior art) Conventionally, optical resolution of 2-alkanol using enzymes and 2-
As a method for synthesizing optically active esters of alkanols,
There is a method for specifically hydrolyzing fatty acid esters of 2-alkanol as shown in the following formula (wherein R11 is a carbon number 1 to
12 alkyl group and R12 represents an alkyl group having 1 to 10 carbon atoms. ) However, enzymes that catalyze this type of reaction are susceptible to the inhibitory effect of fatty acids produced by the reaction, and have the disadvantage that the reaction may stop midway or be delayed. Also known is a method of synthesizing an optically active ester using esterification, which is a reverse reaction of hydrolysis, as shown in the following formula.
脂肪酸
アルコール
（ラセミ体）
光学括性エステル
（式中、Ｒ及びＲ１２は前記と同し意味をもつ。）この
場合も原料である脂肪酸による阻害効果あるいは酵素の
失活を防ぐため濃い緩衝剤を用いられている。また、逆
反応を用いているので、反応の平衡をずらすため一方の
基質（一般にはアルコール）を大過刺に使用して行われ
ており、目的物の分離面てコスト高になるという欠点が
ある。Fatty acid alcohol (racemic) Optical ester (in the formula, R and R12 have the same meanings as above) In this case, too, a strong buffer is used to prevent the inhibitory effect of the raw fatty acid or the deactivation of the enzyme. It is being In addition, since the reverse reaction is used, one substrate (generally alcohol) is used extensively to shift the reaction equilibrium, which has the disadvantage of increasing costs in terms of separation of the target product. be.
これに対して、反応自体に水が関与しないエステル交換
を用いれば多量の脂肪酸を生成させることなしに目的と
する２−アルカノールの光学活性エステルの合成や２−
アルカノールの光学分割か原理的に可能となる。この方
法はすてにＫ１１ｂａｎｏｖら ［［１，Ｃｏｍｂｏ
ｕ、 Ａ、 Ｍ−Ｋｌｉｖａｎｏｖ、 Ｇ、
Ａｍ、 Ｃｈｅｍ。On the other hand, if transesterification is used, which does not involve water in the reaction itself, the desired optically active ester of 2-alkanol can be synthesized without producing a large amount of fatty acid.
In principle, optical resolution of alkanol is possible. This method has been described by K11banov et al. [[1, Combo
u, A, M-Klivanov, G.
Am, Chem.
Ｓｏｃ、、 １．０６．２６８７〜２６９２（１９８４
） ］によって報告されている。彼らの方法は多孔性担
体の孔に酵素を固定化して、多孔体の孔の界面（酵素は
孔の中の緩衝液中に溶解）て行う、いわゆる二相系反応
により光学活性エステルを合成する方法である。Soc, 1.06.2687-2692 (1984
) ] reported by. Their method involves immobilizing an enzyme in the pores of a porous carrier and synthesizing an optically active ester through a so-called two-phase reaction, which is carried out at the pore interface of the porous material (the enzyme is dissolved in a buffer solution within the pores). It's a method.
Ｋ１１ｂａｎｏｖらの行った固定化てはＣａｎｄｉｄａ
ｃｙｌ １ｎｄｒａｃｅａ由来のリパーゼを用いて、１
−リブチリンと数種の２−アルカノールとのエステル交
換を行っているか、本発明者らの追試の結果ｌ）酵素タ
ンパクの孔への固定化の再現性か乏しく、むし多孔性担
体の表面に酵素が付着する。The immobilization performed by K11banov et al.
1 using lipase derived from cyl 1ndracea.
- As a result of follow-up tests conducted by the present inventors, whether or not transesterification of butyrin and several types of 2-alkanols was performed l) The reproducibility of immobilization of the enzyme protein in the pores was poor, and it was found that the enzyme protein was not transesterified on the surface of the porous carrier. is attached.
２）そのため１反応中に酵素タンパクか敲脱し、文献に
示されるような反応は殆ど起こらない（同一条件でのト
リブチリンと２−オクタツールとの反応では高々４〜５
％位てあった）。2) Therefore, the enzyme protein is evaporated during one reaction, and the reaction shown in the literature hardly occurs (in the reaction of tributyrin and 2-octatool under the same conditions, at most 4 to 5
%).
という問題点があることがわかった。It turns out that there is a problem.
（発明か解決しようとする課題）
したかって本発明は加水分解による方法では必然的（主
反応の生成物として）生ずる脂肪酸の阻害効果を除くか
、もしくは軽減し、かつ、固定化などのはん雑な操作な
しに容易に２−アルカノールの光学活性エステルを製造
する方法を提供することを目的とする。(Problem to be Solved by the Invention) Therefore, the present invention eliminates or reduces the inhibitory effect of fatty acids that inevitably occurs (as a product of the main reaction) in hydrolytic methods, and also eliminates the effects of fatty acids such as immobilization. An object of the present invention is to provide a method for easily producing optically active esters of 2-alkanols without complicated operations.
（課題を解決するための手段）
本発明の上記目的は、トリグリセリドと２−アルカノー
ルとを実質的に無水の条件下、粉末状リパーゼの存在下
てエステル交換反応させることを特徴とする２〜アルカ
ノールの光学活性エステルの製造方法により達成された
。(Means for Solving the Problems) The above object of the present invention is to produce a 2-alkanol characterized by transesterifying a triglyceride and a 2-alkanol under substantially anhydrous conditions in the presence of powdered lipase. This was achieved by the method for producing optically active esters.
本発明の反応は次式によって表わすことができる。The reaction of the present invention can be represented by the following formula.
（式中、Ｒ１及びＲ２はアルキル基を示す。）本発明に
用いられるトリグリセリド（Ｚ）においてＲ１のアルキ
ル基の炭素数は特に制限はないが、３以上か好ましく、
３〜２２かより好ましい。このアルキル基の炭素数か大
きくなるとトリグリセリドは常温で固体となるが、有機
溶媒中て反応を行わせることによりその反応に使用する
ことかてきる。(In the formula, R1 and R2 represent an alkyl group.) In the triglyceride (Z) used in the present invention, the number of carbon atoms in the alkyl group of R1 is not particularly limited, but it is preferably 3 or more,
3 to 22 is more preferable. When the number of carbon atoms in this alkyl group increases, the triglyceride becomes solid at room temperature, but it can be used in the reaction by carrying out the reaction in an organic solvent.
また式（ＩＩ ）で表わされる２−アルカノールにおけ
るＲ２て表わされるアルキル基の炭素数は通常１〜３０
てあり、好ましくは３〜１２である。Furthermore, the number of carbon atoms in the alkyl group represented by R2 in the 2-alkanol represented by formula (II) is usually 1 to 30.
and preferably 3 to 12.
本発明の反応は実質的に無水の条件下で行われるが、実
質的に無水とは含有水分量か１ｖｏ１％以下のことをい
い、これは無溶媒又は非水溶媒中での反応させることに
より達成てきる。このように実質的に無水で反応させる
ことにより副反応（加水分解）による脂肪酸の生成を防
ぐことかてきる。The reaction of the present invention is carried out under substantially anhydrous conditions, and "substantially anhydrous" refers to a water content of 1 vol. I can achieve it. By performing the reaction in a substantially anhydrous state in this way, it is possible to prevent the production of fatty acids due to side reactions (hydrolysis).
本発明の反応に用いられる非水溶媒（有機溶媒）として
はヘキサン、ヘプタン、オクタン、イソオクタン、四塩
化炭宋、イソプロピルエーテル、シーｎ−ブチルエーテ
ルなどが挙げられる。Examples of the non-aqueous solvent (organic solvent) used in the reaction of the present invention include hexane, heptane, octane, isooctane, carbon tetrachloride, isopropyl ether, and n-butyl ether.
本発明に用いられるリパーゼとして、ポルシン０バンク
レアス（Ｐｏｒｃｉｎ ｐａｎｃｒｅａｓ） 、シュー
トモナース・フローレスセンス（Ｐｓｅｕｄｏｍｏｎａ
ｓｆｌｕｏｒｅｓｃｅｎｓ） 、シュートモナース・ス
ベーシーズ（Ｐｓｅｕｄｏｍｏｎａｓ ５ｐｅｃｉｅｓ
）か好ましいものとして挙げられる。As the lipase used in the present invention, Porcin pancreas, Pseudomonas florescens
sfluorescens), Pseudomonas 5pecies
) are listed as preferred.
このリパーゼは粉末状で反応系に添加されるが、リパー
ゼは粉末状態のまま加え、通常の化学反応で用いられる
ような攪拌下で行う、この場合、コストを下げるため、
酵素は安価で入手可能な粗精製品を用いることができる
。This lipase is added to the reaction system in powder form.In this case, in order to reduce costs, lipase is added in powder form and the process is carried out under stirring as used in ordinary chemical reactions.
As the enzyme, crude products that are available at low cost can be used.
リパーゼの使用量は、反応溶液１ｍｌに対し通常１０ｏ
＋ｇ以上、より好ましくは５０〜３００ｍｇの範囲であ
る。The amount of lipase used is usually 10o per 1ml of reaction solution.
+g or more, more preferably in the range of 50 to 300 mg.
トリグリセリド（Ｉ）と２−アルカノール（ｎ）とのモ
ル比（ＩＩ ） ／ ＣＩ ）は通常１〜２０、好まし
くは１〜３である。The molar ratio (II)/CI) of triglyceride (I) and 2-alkanol (n) is usually 1 to 20, preferably 1 to 3.
反応温度は１０〜９０℃、好ましくは常温であり、反応
時間は５時間以上、好ましくは１０時間以上、より好ま
しくは３０〜１５０時間である。The reaction temperature is 10 to 90°C, preferably room temperature, and the reaction time is 5 hours or more, preferably 10 hours or more, and more preferably 30 to 150 hours.
本発明において目的の光学活性エステルの反応系からの
分離は常法に従って行うことができる。In the present invention, the target optically active ester can be separated from the reaction system according to a conventional method.
具体的には減圧蒸留、カラムクロマトグラフィーなどを
採用して行うことができる。Specifically, this can be carried out by employing vacuum distillation, column chromatography, or the like.
（発明の効果）
本発明によれば、反応は無溶媒又は非水溶媒中で行われ
脂肪酸等の生成を経ることなく、選択率良く、トリグリ
セリドから２−アルカノールの光学活性エステルを得る
ことができるという優れた効果を奏する。この際、酵素
（リパーゼ）の固定化などのはん雑な操作を必要とせず
、工業的に実施する方法として好適である。(Effects of the Invention) According to the present invention, optically active esters of 2-alkanols can be obtained from triglycerides with good selectivity because the reaction is carried out without a solvent or in a non-aqueous solvent and without producing fatty acids or the like. It has this excellent effect. At this time, it does not require complicated operations such as immobilization of enzyme (lipase), and is suitable as a method to be implemented industrially.
さらに本発明方法は有機溶媒中て行う場合ても酵素の活
性が発現されるばかりでなく常温で液体のトリグリセリ
ドの代わりに常温て固体のもの（例えばトリラウリン）
を原料として利用でき、原料選択の幅か広いという優れ
た効果を奏する。Furthermore, even when the method of the present invention is carried out in an organic solvent, not only the enzyme activity is expressed, but also triglyceride which is solid at room temperature (for example, trilaurin) is used instead of triglyceride which is liquid at room temperature.
can be used as a raw material, and has the excellent effect of widening the range of raw material selection.
本発明方法により得られる光学活性エステルは医薬品の
中間体である光学活性２−アルカノールの製造、光学活
性液晶材料の製造などに有用である。The optically active ester obtained by the method of the present invention is useful in the production of optically active 2-alkanol, which is an intermediate for pharmaceuticals, and in the production of optically active liquid crystal materials.
（実施例） 次に本発明を実施例に基づきさらに詳細に説明する。(Example) Next, the present invention will be explained in more detail based on examples.
実施例ｌ
Ｒ３−２−オクタツール０．２Ｍを含むトリブチリンｌ
ｏｄと第１表に示す酵素１．０ｇを丸底フラスコに採り
さらに反応液所定量の水を添加した。次に、３０℃でフ
ットボール型攪拌子により５００ｒｐｍ以上で回転させ
ながら反応液を攪拌した。２４時間後、反応液の一部（
０，Ｓｌ）を５ｍｌのクロロホルムに希釈し、濾過で酵
素を除き、ガスクロマトグラフィーで分析を行った。結
果を第２表に示した。Example l Tributyrin l containing 0.2M R3-2-octatool
od and 1.0 g of the enzyme shown in Table 1 were placed in a round bottom flask, and a predetermined amount of water was added to the reaction solution. Next, the reaction solution was stirred at 30° C. while rotating at 500 rpm or more using a football-type stirrer. After 24 hours, a portion of the reaction solution (
0.0, Sl) was diluted in 5 ml of chloroform, the enzyme was removed by filtration, and analysis was performed by gas chromatography. The results are shown in Table 2.
第１表 酵素（リパーゼ）の略号。Table 1 Abbreviations of enzymes (lipase).
起源及び酵素活性
略号 起 源 活性８ＩＣ，ｃ、
Ｃａｎｄｉｄａ ｃｙｌｉｎｄｒａｃｅａ
６０Ｐ、ｓ、 Ｐｓｅｕｄｏｍｏｎａｓ ５ｐ
ｅｃｉｅｓ ３３Ｐ、ｌ Ｐｓｅｕｄｏｗ
ｏｎａｓ ｆｌｕｏｒｅｓｃｅｎｓ ３２Ｐ、ｐ、
Ｐｏｒｃｉｎｅ ｐａｎｃｒｅａｓ
１１Ｐ、ｃ、 Ｐｅｎｉｃｉｌｌｉｕｍ ｃｙｃｌ
ｓｐｉｕｍ ２０注）業ｌオリーブ油の加水分解
から求めたもの。Origin and enzyme activity abbreviation Origin Activity 8IC, c,
Candida cylindracea
60P, s, Pseudomonas 5p
ecies 33P,l Pseudow
onas fluorescens 32P, p,
Porcine pancreas
11P, c, Penicillium cycle
Spium 20 Note) Obtained from the hydrolysis of olive oil.
単位：用／ ｍ ｇ
第２表 トリブチリン中、リパーゼによるＲ３−２−オ
クタノールとトリブチリンとの
エステル交換反応率１）
１）Ｒ３−２−オクタノールを基準とした反応率を示す
。Units: /mg Table 2 Transesterification reaction rate of R3-2-octanol and tributyrin in tributyrin by lipase 1) 1) The reaction rate is shown based on R3-2-octanol.
２）酵素タンパクの凝集か起こり、攪拌か不良てあった
。なお、添加水分量が５．１０％の時はすべての場合に
酵素タンパクの凝集が認められた。2) Enzyme protein aggregation occurred, and stirring was inadequate. Incidentally, when the amount of added water was 5.10%, aggregation of enzyme protein was observed in all cases.
第２表の結果より明らかなようにＣ，ｃ、リパーゼでは
添加水分量が０．５％の時に最大の活性を示すが、他の
三種のリパーゼでは添加水分量が０％のときに最大活性
（反応率）を示す。また、いずれの場合も、添加水分量
か１％を越えると極端に反応性が低下する。As is clear from the results in Table 2, C, C, and lipases exhibit maximum activity when the amount of added water is 0.5%, while the other three types of lipase have maximum activity when the amount of added water is 0%. (reaction rate). Furthermore, in any case, if the amount of added water exceeds 1%, the reactivity is extremely reduced.
上記の実験とそれぞれ同様の反応条件て、添加水分量の
効果を詳しく調べた。本実験では特に反応初期について
調べた。結果を第１図に示す。The effect of the amount of added water was investigated in detail under the same reaction conditions as in the above experiments. In this experiment, we particularly investigated the initial stage of the reaction. The results are shown in Figure 1.
リパーゼの種類と添加水分量Ｏ％のときの反応溶液中の
水分量は次の通りである。The type of lipase and the amount of water in the reaction solution when the amount of added water is 0% are as follows.
Ａ ： Ｃ，ｃ、（０，：１４ｖｏｌＸ） Ｂ ：
Ｐ、ｓ、（０，２：１ｖｏｌ＄）Ｃ： Ｐ、ｆ、（０，
１７ｖｏｌＸ） Ｄ ： Ｐ、ｐ、（０，０７ｖｏｌ
りＥ ： Ｐ、ｃ、（０，１４ｖｏｌ＄）反応時間（１
）は次の通りである。A: C, c, (0,:14volX) B:
P, s, (0, 2: 1 vol $) C: P, f, (0,
17volX) D: P, p, (0,07vol
Reaction time (1
) is as follows.
ａ ： ｔ＝３０分 ｂ ： ｔ＝６５分 Ｃ：
ｔ＝ １時間ｄ ： ｔ＝ ２時間 ｅ ： ｔ＝ ３
時間 ｆ ： ｔ＝ ５時間ｇ ： ｔ＝１０時間 ｈ
： ｔ−２３時間第１図の結果から明らかなようにＣ
，ｃ、リパーゼでは添加水分量か０．５ｖｏ１％のとき
に最大の活性を示すか、他の５種の酵素ては添加水分量
か０％のときに最大の活性を示し、添加水分量か増加す
ると酵素活性が低下する。a: t=30 minutes b: t=65 minutes C:
t= 1 hour d: t= 2 hours e: t= 3
Time f: t=5 hours g: t=10 hours h
: t-23 hours As is clear from the results in Figure 1, C
, c, lipase shows maximum activity when the amount of added water is 0.5 vol 1%, and the other five enzymes show maximum activity when the amount of added water is 0%, and the amount of added water shows maximum activity. As it increases, enzyme activity decreases.
実施例２
Ｒ（−）−２−才クタノール０．２ＭまたはＳ （＋）
−２−オクタツール０．２Ｍを含むトリブチリン溶液１
０ｍ１に酵素１ｇを加え（Ｃ，ｃ、の場合のみ水を０．
５ｖｏ１％＝５０ｐ、！；Ｌを添加した）、３０°ＣＣ
１５００ｒｐ以上で攪拌しながら反応を行った。各反応
の反応率を１０時間にわたって測定した結果を第２図に
示した。Example 2 R(-)-2-year-old ethanol 0.2M or S(+)
-Tributyrin solution 1 containing 2-octatool 0.2M
Add 1 g of enzyme to 0 ml (only in case of C, c, add 0.0 ml of water).
5vo1%=50p,! ;L was added), 30°C
The reaction was carried out while stirring at 1500 rpm or more. The reaction rate of each reaction was measured over 10 hours and the results are shown in FIG.
第２図の結果より、明らかなように、Ｐ、ｐ、、Ｐ、ｓ
、、 Ｐ、ｆ、リパーゼの場合はＲ（−） −２−オク
タノールの方がＳ （＋） −２−オクタツールよりも
かなり速く反応する。すなわち、Ｒ３−２−オクタツー
ルとトリブチリンとのエステル交換ではＲ（−）−２−
オクタノールのエステルを優先的に生成することかわか
る。From the results in Figure 2, it is clear that P, p, , P, s
,, P, f, In the case of lipase, R(-)-2-octanol reacts much faster than S(+)-2-octatool. That is, in the transesterification of R3-2-octatool and tributyrin, R(-)-2-
It can be seen that esters of octanol are preferentially produced.
一方、Ｐ、ｃ、、Ｐ、乙、リパーゼの場合は１）、２）
とは逆に８体の方がＲ体よりも反応か速く、ラセミ体を
用いた合成ては８体アルコールのエステルか多く発生す
ることかわかる。On the other hand, in the case of P, c, , P, Otsu, and lipase, 1), 2)
On the contrary, the reaction of the 8-isomer is faster than that of the R-isomer, and it can be seen that more esters of the 8-isomer alcohol are generated in the synthesis using the racemate.
実施例３
ＲＳ−２−オクタツール２４ｇとトリブチリン７０ｇと
の混合物に酵素（Ｐ、ｆ、リパーゼ）１０ｇを加え、３
０℃、５００ｒｐｍ以上で攪拌して反応させた。５０時
間後１反応液を３００ｍ１のヘキサンに溶解させ、濾過
により酵素タンパクを除去し、無水硫酸ナトリウムで脱
水し、エバポレーターで溶媒を除去した。残留物の重量
は９１．６ｇてあった。この残留物を減圧蒸留し、７７
〜ｂ
テルか主成分）を合わせ、シリカゲル（２４０ｇ）を充
填したカラムに負荷した。５０％ベンゼン／ヘキサンて
溶出すると０．４〜１．６１のフラクションから目的と
する光学活性２−オクチル酪酸（純度１００％）が得ら
れた。Example 3 10 g of enzymes (P, f, lipase) were added to a mixture of 24 g of RS-2-octatool and 70 g of tributyrin, and 3
The reaction was carried out at 0° C. and stirred at 500 rpm or more. After 50 hours, one reaction solution was dissolved in 300 ml of hexane, the enzyme protein was removed by filtration, dehydrated with anhydrous sodium sulfate, and the solvent was removed with an evaporator. The weight of the residue was 91.6 g. This residue was distilled under reduced pressure and
-b (main component) were combined and loaded onto a column packed with silica gel (240 g). When eluted with 50% benzene/hexane, the desired optically active 2-octylbutyric acid (purity 100%) was obtained from fractions 0.4 to 1.61.
Ｐ、 ｆ、以外の酵素も同様の方法で、目的とするエス
テルを得た。結果を第３表に示した。The desired esters were obtained using enzymes other than P and f in the same manner. The results are shown in Table 3.
第３表 酵素（リパーゼ）を用いたＲ３−２−オクタツ
ールとトリブチリンとのエステル交換による光学エステ
ルの合成
（注）１）Ｒ３−２−オクタノールに対する反応率を示
す。Table 3 Synthesis of optical ester by transesterification of R3-2-octatool and tributyrin using an enzyme (lipase) (Note 1) Shows the reaction rate for R3-2-octanol.
２）反応率から計算した理論量に対する純度１００％の
エステルの収率
３） ＲはＲ（−）−２−オクチル酪酸、ＳはＳ （＋
） −２−オクチル酪酸を示す。2) Yield of 100% pure ester based on the theoretical amount calculated from the reaction rate 3) R is R(-)-2-octylbutyric acid, S is S (+
) -2-octylbutyric acid.
また、光学純度は下式から計算した。Moreover, optical purity was calculated from the following formula.
ここで、標品の［α］、 は第３表のＲの場合（Ｐ、ｓ
、、Ｐ、 ｆ、、Ｐ、ｐ、 ）はＲ（−） −２−オク
タノールから合成した醋酸エステルの［α］。Here, [α] of the standard is for R in Table 3 (P, s
, , P, f, , P, p, ) is [α] of acetic acid ester synthesized from R(-)-2-octanol.
を示す。shows.
第３表の結果から明らかなようにＰ、ｓ、、Ｐ、ｆ、、
ｐ、ｐ、リパーゼを用いてＲ（−） −２−オクチル酪
酸（光学活性エステル）か得られ、反応は遅いかｐ、ｐ
、が最も優れており、Ｐ、ｐ、＞Ｐ、ｓ、＞Ｐ、ｆ、の
順の特異性を示す。As is clear from the results in Table 3, P, s, , P, f, .
R(-)-2-octylbutyric acid (optically active ester) is obtained using p, p, lipase, and the reaction is slow or p, p
, is the best, and shows specificity in the order of P, p, > P, s, > P, f.
実施例４
２−オクタツール以外の２−アルカノールを用いた以外
は実施例３と同様にしてエステル交換による対応光学活
性エステルの合成を行った。その結果を第４表に示した
。Example 4 A corresponding optically active ester was synthesized by transesterification in the same manner as in Example 3 except that 2-alkanol other than 2-octatool was used. The results are shown in Table 4.
実施例５
Ｒもしくは５−２−オクタツール０．２Ｍとトリブチリ
ン０．２Ｍとの混合物を下記第５表に示す溶媒に溶解し
リパーゼを１００ｍｇ／Ｗｉｇの濃度て加え、水分量の
０．２〜０．６ｖｏ １％で、３０℃、５００ｒｐｍ以
上で攪拌してそれぞれの反応の経時変化を調べた。以下
実施例２と同様に処理して目的とするエステルの生成率
を求めた。Example 5 A mixture of 0.2 M of R or 5-2-octatool and 0.2 M of tributyrin was dissolved in the solvent shown in Table 5 below, and lipase was added at a concentration of 100 mg/Wig to reduce the water content from 0.2 to The reaction mixture was stirred at 0.6vo 1% at 30° C. and at 500 rpm or higher to examine changes over time in each reaction. Thereafter, the same procedure as in Example 2 was carried out to determine the production rate of the desired ester.
その結果を第５表に示した。The results are shown in Table 5.
同表から明らかなようにＲ体及び８体アルコールの一定
時間後の反応率から［Ｒ（％）−８（％）］／［Ｒ（％
）＋Ｓ（％）コを計算すると、はとんど一定となった。As is clear from the same table, from the reaction rate of R- and 8-alcohols after a certain period of time, [R (%) - 8 (%)] / [R (%
) + S (%) was calculated, and it became almost constant.
ここでＲ（％）及びＳ（％）は一定時間後（いずれも同
一時間）の反応率を示す。また［Ｒ（％）−８（％）コ
／［Ｒ（％）＋Ｓ（％）コはＲ５−２−オクタツールと
トリブチリンとのエステル交換から合成した光学活性エ
ステル、光学純度（ｅ、ｅ、）に相当する０例えばトリ
ブチリン中、Ｐ、ｓ、リパーゼによるエステル交換ては
［Ｒ（％）−８（％）］／［Ｒ（％）＋Ｓ（％）］の値
は０．８０〜０．８３てあり、単離したエステルのｅ、
ｅ、は０．８１とほぼ一致する。また、ｐ、ｐ、てはＡ
＝ ［Ｒ，Ｃ％）−３（％）］／［Ｒ（％）＋Ｓ（％）
］は０．８７で、ｅ、ｅ、は０．９９以上であり、単離
エステルの方かＡの値から予想するよりも高いｅ、ｅ−
を有することになる。したかって、第５表からは、有機
溶媒中では［Ｒ（％） −３（％） コ ／ ［Ｒ（％
） ＋Ｓ （％） ］と同程度か、それ以上のｅ、ｅ、
（光学純度）を有するエステルか合成てきることが分
る。Here, R (%) and S (%) indicate the reaction rate after a certain time (both are the same time). In addition, [R (%) - 8 (%) co/[R (%) + S (%) co] is an optically active ester synthesized by transesterification of R5-2-octatool and tributyrin, and the optical purity (e, e, ) corresponds to 0. For example, in tributyrin, P, s, transesterification with lipase, the value of [R (%) - 8 (%)] / [R (%) + S (%)] is 0.80 to 0. 83 and the isolated ester e,
e, is approximately equal to 0.81. Also, p, p, te is A
= [R,C%)-3(%)]/[R(%)+S(%)
] is 0.87, e, e, is 0.99 or more, and e, e- is higher than expected from the value of A for the isolated ester.
will have the following. Therefore, from Table 5, in organic solvents, [R(%) -3(%) / [R(%)
) +S (%) ] or greater e, e,
It turns out that esters with high optical purity can be synthesized.
仇５表 リパーゼによる２−オクタツールの［Ｒ（％）
−３（％）］／［Ｒ（％）＋Ｓ（％）］／Table 5: [R (%)] of 2-octatool by lipase
−3(%)]/[R(%)+S(%)]/
ｆ５１［Ｚ（Ａ）〜（Ｅ）はエステル交換反応率と反応
系の含有水分量との関係を示すグラフである。第２図Ｒ
（＝）又はＳ （＋） −２−オクタツールのエステル
交換反応率をそれぞれ示すグラフである。
ａ）基質濃度：２−オクタツール（Ｒあるいは３体）０
．２０Ｍ、ＴＢ ０．２０Ｍ。
酵素濃度； １００ ｍｇ／ｍ！ｌ！。
反応温度；３０°Ｃ１
水分ｌ； ０．２〜０．６ ｖｏ１％
ｂ）いずれもＲ体アルコールの方が早く、符号は正であ
った。
第
図
０．５
添加した水分量
（容量比。
％）
第
図（その１）
反応時間（５＋間）f51[Z(A) to (E) are graphs showing the relationship between the transesterification reaction rate and the water content of the reaction system. Figure 2 R
(=) or S (+) - It is a graph which shows the transesterification reaction rate of -2-octatool, respectively. a) Substrate concentration: 2-octatool (R or 3 form) 0
．． 20M, TB 0.20M. Enzyme concentration: 100 mg/m! l! . Reaction temperature: 30°C1 Moisture 1: 0.2-0.6 vol% b) In both cases, the R-form alcohol was faster and the sign was positive. Figure 0.5 Amount of water added (volume ratio. %) Figure (Part 1) Reaction time (5+ minutes)
Claims (1)
水の条件下、粉末状リパーゼの存在下でエステル交換反
応させることを特徴とする２−アルカノールの光学活性
エステルの製造方法。1. A method for producing an optically active ester of 2-alkanol, which comprises transesterifying triglyceride and 2-alkanol under substantially anhydrous conditions in the presence of powdered lipase.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP63239373A JPH0634752B2 (en) | 1988-09-24 | 1988-09-24 | Method for producing optically active ester of 2-alkanol |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP63239373A JPH0634752B2 (en) | 1988-09-24 | 1988-09-24 | Method for producing optically active ester of 2-alkanol |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JPH0286797A true JPH0286797A (en) | 1990-03-27 |
| JPH0634752B2 JPH0634752B2 (en) | 1994-05-11 |
Family
ID=17043809
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP63239373A Expired - Lifetime JPH0634752B2 (en) | 1988-09-24 | 1988-09-24 | Method for producing optically active ester of 2-alkanol |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPH0634752B2 (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH07303494A (en) * | 1994-03-15 | 1995-11-21 | Nisshin Oil Mills Ltd:The | Production of optically active alcohol |
| EP0709465A2 (en) | 1994-10-26 | 1996-05-01 | The Nisshin Oil Mills, Ltd. | Optical resolution for producing optically active alcohol |
| US5600027A (en) * | 1994-11-29 | 1997-02-04 | The Nisshin Oil Mills, Ltd. | Process for producing optically active alcohol containing phenyl group |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS62166898A (en) * | 1986-01-21 | 1987-07-23 | Chisso Corp | Production of optically active alcohol by biochemical technique |
-
1988
- 1988-09-24 JP JP63239373A patent/JPH0634752B2/en not_active Expired - Lifetime
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS62166898A (en) * | 1986-01-21 | 1987-07-23 | Chisso Corp | Production of optically active alcohol by biochemical technique |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH07303494A (en) * | 1994-03-15 | 1995-11-21 | Nisshin Oil Mills Ltd:The | Production of optically active alcohol |
| EP0709465A2 (en) | 1994-10-26 | 1996-05-01 | The Nisshin Oil Mills, Ltd. | Optical resolution for producing optically active alcohol |
| EP0709465A3 (en) * | 1994-10-26 | 1997-04-23 | Nisshin Oil Mills Ltd | Optical resolution for producing optically active alcohol |
| US5696299A (en) * | 1994-10-26 | 1997-12-09 | The Nisshin Oil Mills, Ltd. | Optical resolution for producing optically active alcohol |
| US5600027A (en) * | 1994-11-29 | 1997-02-04 | The Nisshin Oil Mills, Ltd. | Process for producing optically active alcohol containing phenyl group |
Also Published As
| Publication number | Publication date |
|---|---|
| JPH0634752B2 (en) | 1994-05-11 |
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