JPS62205797A - Production of optically active compound - Google Patents
Production of optically active compoundInfo
- Publication number
- JPS62205797A JPS62205797A JP4790386A JP4790386A JPS62205797A JP S62205797 A JPS62205797 A JP S62205797A JP 4790386 A JP4790386 A JP 4790386A JP 4790386 A JP4790386 A JP 4790386A JP S62205797 A JPS62205797 A JP S62205797A
- Authority
- JP
- Japan
- Prior art keywords
- acid
- optically active
- ester
- halocarboxylic
- active compound
- 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
- 238000004519 manufacturing process Methods 0.000 title claims description 7
- 150000001875 compounds Chemical class 0.000 title claims description 5
- 150000002148 esters Chemical class 0.000 claims abstract description 16
- 108090000790 Enzymes Proteins 0.000 claims abstract description 14
- 102000004190 Enzymes Human genes 0.000 claims abstract description 14
- 241000223198 Humicola Species 0.000 claims abstract description 7
- 125000000217 alkyl group Chemical group 0.000 claims abstract description 3
- 125000003710 aryl alkyl group Chemical group 0.000 claims abstract description 3
- 125000000113 cyclohexyl group Chemical group [H]C1([H])C([H])([H])C([H])([H])C([H])(*)C([H])([H])C1([H])[H] 0.000 claims abstract description 3
- 125000004417 unsaturated alkyl group Chemical group 0.000 claims abstract description 3
- 244000005700 microbiome Species 0.000 claims description 5
- 150000001721 carbon Chemical class 0.000 claims 1
- 239000002253 acid Substances 0.000 abstract description 10
- 108090001060 Lipase Proteins 0.000 abstract description 8
- 239000004367 Lipase Substances 0.000 abstract description 8
- 102000004882 Lipase Human genes 0.000 abstract description 8
- 235000019421 lipase Nutrition 0.000 abstract description 8
- -1 alpha-halocarboxylic acid ester Chemical class 0.000 abstract description 7
- 238000006460 hydrolysis reaction Methods 0.000 abstract description 4
- 150000003978 alpha-halocarboxylic acids Chemical class 0.000 abstract description 2
- 150000001732 carboxylic acid derivatives Chemical class 0.000 abstract description 2
- 125000005843 halogen group Chemical group 0.000 abstract description 2
- 230000000813 microbial effect Effects 0.000 abstract description 2
- 229920006395 saturated elastomer Polymers 0.000 abstract description 2
- 230000007062 hydrolysis Effects 0.000 abstract 2
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 description 12
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 6
- 238000006243 chemical reaction Methods 0.000 description 5
- 238000000034 method Methods 0.000 description 5
- 230000003287 optical effect Effects 0.000 description 4
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 description 2
- 239000003795 chemical substances by application Substances 0.000 description 2
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 description 2
- 239000010410 layer Substances 0.000 description 2
- BGMYDZVCAOVIPM-UHFFFAOYSA-N octyl 2-chloropropanoate Chemical compound CCCCCCCCOC(=O)C(C)Cl BGMYDZVCAOVIPM-UHFFFAOYSA-N 0.000 description 2
- 239000008363 phosphate buffer Substances 0.000 description 2
- 239000000243 solution Substances 0.000 description 2
- 108090000371 Esterases Proteins 0.000 description 1
- 239000003905 agrochemical Substances 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
- 239000000872 buffer Substances 0.000 description 1
- KATNUXHENWPMQJ-LURJTMIESA-N butyl (2s)-2-chloropropanoate Chemical compound CCCCOC(=O)[C@H](C)Cl KATNUXHENWPMQJ-LURJTMIESA-N 0.000 description 1
- PMVCCUUEVYCMBN-UHFFFAOYSA-N butyl 2-chloropentanoate Chemical compound CCCCOC(=O)C(Cl)CCC PMVCCUUEVYCMBN-UHFFFAOYSA-N 0.000 description 1
- KATNUXHENWPMQJ-UHFFFAOYSA-N butyl 2-chloropropanoate Chemical compound CCCCOC(=O)C(C)Cl KATNUXHENWPMQJ-UHFFFAOYSA-N 0.000 description 1
- 125000004432 carbon atom Chemical group C* 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 239000012141 concentrate Substances 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 230000002255 enzymatic effect Effects 0.000 description 1
- 238000006911 enzymatic reaction Methods 0.000 description 1
- 230000003301 hydrolyzing effect Effects 0.000 description 1
- 239000004310 lactic acid Substances 0.000 description 1
- 235000014655 lactic acid Nutrition 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 239000012044 organic layer Substances 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 230000035484 reaction time Effects 0.000 description 1
- 238000001953 recrystallisation Methods 0.000 description 1
- 238000004904 shortening Methods 0.000 description 1
- 238000003756 stirring Methods 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
Landscapes
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
Description
【発明の詳細な説明】
〔産業上の利用分野〕
本発明は光学活性α−ハロカルボン酸エステル及びその
対掌体α−ハロカルボン酸の製造法に関する。更に詳し
くはフミコラ（Ｈｕｍｉｃｏｌａ）属に属する微生物に
由来する酵素で、一般式（Ｉ）（Ｒは炭素数が１から２
０の飽和または不飽和のアルキル基を、Ｒ゛は直鎖ある
いは分岐のアルキル基、アラルキル基又はシクロヘキシ
ル基を、またＸはハロゲン原子を示す。）
で表されるα−ハロカルボン酸エステルのラセミ体を不
斉加水分解した後、生成する酸と残存するエステルを分
離・採取する光学活性化合物の製造法に関する。DETAILED DESCRIPTION OF THE INVENTION [Field of Industrial Application] The present invention relates to a method for producing an optically active α-halocarboxylic acid ester and its enantiomer α-halocarboxylic acid. More specifically, it is an enzyme derived from a microorganism belonging to the Humicola genus, and has the general formula (I) (R has 1 to 2 carbon atoms).
0 represents a saturated or unsaturated alkyl group, R′ represents a linear or branched alkyl group, aralkyl group or cyclohexyl group, and X represents a halogen atom. The present invention relates to a method for producing an optically active compound, which involves asymmetrically hydrolyzing a racemic form of an α-halocarboxylic acid ester represented by the following formula, and then separating and collecting the generated acid and the remaining ester.
一般式（１）で表されるエステルの光学活性体又はその
対掌体である光学活性な酸は各挿図・農薬の中間体とし
て重要な化合物である。The optically active form of the ester represented by the general formula (1) or the optically active acid which is its antipode is an important compound as an intermediate for each of the illustrations and agricultural chemicals.
前記一般式（Ｉ）で示されるエステルの光学活性体又は
その対掌体である光学活性な酸の製造法としては、従来
、光学活性な乳酸をエステル化した後にハロゲン化する
方法、α−ハロカルボン酸を光学分割剤を用いて光学分
割する方法等が知られている。しかし、これらの方法は
高価な光学活性体化合物を原料としたり、あるいは高価
な光学分割試薬を用いての再結晶を繰り返さなければな
らず、必ずしも工業的方法として優れたものとは言い難
い。Conventionally, methods for producing an optically active form of the ester represented by the general formula (I) or an optically active acid which is its enantiomer include a method in which optically active lactic acid is esterified and then halogenated, and α-halocarbon A method of optically resolving an acid using an optical resolving agent is known. However, these methods require expensive optically active compounds as raw materials or require repeated recrystallization using expensive optical resolution reagents, and cannot necessarily be said to be excellent as industrial methods.
一方、酵素を用いる製造法に関しても、クリバッフ（Ａ
ｐｐｌ、Ｂｉｏｃｈｅｍ、ａｎｄ Ｂｉｏｔｅｃｈ、９
．２５５（１９８４）） 、入内島ら（特開昭５７−９
４２９５号公報）の報告が知られている。しかし、これ
らで示される酵素の反応でも未だ収率や純度面で十分工
業的に使用しうる方法には到っていないのが実状である
。On the other hand, regarding production methods using enzymes, Kuribafu (A
ppl, Biochem, and Biotech, 9
．． 255 (1984)), Iriuchijima et al.
4295) is known. However, the reality is that even the enzymatic reactions described above have not yet reached a method that can be used industrially in terms of yield and purity.
かかる事情に鑑み、本発明者らは前記一般式（Ｉ）で表
されるエステルの光学活性体又はその対掌体である光学
活性な酸を工業的に製造すべく鋭意検討を行った。その
結果、一般式（Ｉ）で表されるエステルを加水分解し得
るリパーゼ、エステラーゼについて広く検索を行い、フ
ミコラ属に属する微生物から得られる酵素（リパーゼ）
が、一般式（Ｉ）で表されるエステルを、極めて短時間
に効率良くかつ高い光学選択性をもって不斉加水分解す
る事を見出し、本発明をなすにいたった。In view of these circumstances, the present inventors conducted intensive studies to industrially produce an optically active form of the ester represented by the general formula (I) or an optically active acid that is its enantiomer. As a result, we conducted a wide search for lipases and esterases that can hydrolyze the ester represented by the general formula (I), and found that enzymes (lipases) obtained from microorganisms belonging to the genus Humicola.
However, it was discovered that the ester represented by the general formula (I) can be asymmetrically hydrolyzed efficiently in an extremely short period of time and with high optical selectivity, and the present invention has been completed.
即ち、本発明は化学合成で得られる一般式（Ｉ）で示さ
れるα−ハロカルボン酸エステルのラセミ体に、フミコ
ラ属に属する微生物の生産する酵素リパーゼを作用させ
ることにより、不斉加水分解した後、生成する酸と残存
するエステルとを分離・採取することを特徴とする光学
活性なα−ハロカルボン酸エステル又はその対掌体の光
学活性なカルボン酸を製造する方法を提供するものであ
る。That is, the present invention provides a racemic form of α-halocarboxylic acid ester represented by the general formula (I) obtained by chemical synthesis, which is asymmetrically hydrolyzed by causing lipase, an enzyme produced by a microorganism belonging to the genus Humicola, to act on it. The present invention provides a method for producing an optically active α-halocarboxylic acid ester or an optically active carboxylic acid enantiomer thereof, which comprises separating and collecting the generated acid and the remaining ester.
本発明において用いられる酵素はフミコラ属に属する微
生物に由来し、エステルを不斉加水分解する能力を有す
るものであり、好適な例として天野製薬■製“リパーゼ
ＣＢ”を挙げることができる。これら酵素は精製３素、
粗酵素、酵素剤としての微生物菌体あるいは培養液いず
れの形態でも必要に応じて用いることができる。The enzyme used in the present invention is derived from a microorganism belonging to the genus Humicola and has the ability to asymmetrically hydrolyze esters, and a preferred example is "Lipase CB" manufactured by Amano Pharmaceutical Co., Ltd. These enzymes are purified three elements,
Any form of crude enzyme, microbial cells as an enzyme agent, or culture solution can be used as required.
本発明に用いられる一般式（Ｉ）で表されるエステルと
しては、例え、ばα−クロロプロピオン酸−ｎ−ブチル
エステル、α−ブロモプロピオン１−ｎ−７’チルエス
テル、α−クロロプロピオンｆＪｌ−ｎ−オクチルエス
テル、α−クロロ吉草酸−ｎ−ブチルエステル等を挙げ
ることができる。Examples of the ester represented by the general formula (I) used in the present invention include α-chloropropionic acid-n-butyl ester, α-bromopropion 1-n-7′ thyl ester, α-chloropropion fJl- Examples include n-octyl ester and α-chlorovaleric acid n-butyl ester.
加水分解反応を行うに際しては、反応液のｐＨを用いる
酵素の最適ｐｌ＋に合わせてやる必要があり、このため
には緩衝液を使用してもよいし、Ｎ ａ ０１１、Ｋ０
１１水溶液を用いてのｐＨスタットにより反応中のｐＨ
をコントロールしてもよい。反応温度も用いる酵素によ
り異なるが、通常１０〜６０”Ｃの範囲で行われ、好ま
しくは２５〜５０°Ｃで行われる。When carrying out a hydrolysis reaction, it is necessary to adjust the pH of the reaction solution to the optimum pl+ of the enzyme used, and for this purpose a buffer may be used, and Na 011, K0
11 pH during reaction by pH stat using aqueous solution
may be controlled. The reaction temperature also varies depending on the enzyme used, but is usually carried out in the range of 10 to 60"C, preferably 25 to 50"C.
フミコラ属由来の酵素は一般に耐熱性に優れており、例
えばリパーゼＣＥは５０℃でも十分酵素活性を有してい
る。これは加水分解反応時間を短縮するのに極めて都合
がよい。反応は攪拌又は振盪条件下数時間乃至数十時間
行う。Enzymes derived from the genus Humicola generally have excellent heat resistance; for example, lipase CE has sufficient enzymatic activity even at 50°C. This is very convenient for shortening the hydrolysis reaction time. The reaction is carried out under stirring or shaking conditions for several hours to several tens of hours.
反応終了後は通常の方法により生成した酸と残存するエ
ステルを分離・採取することができる。After the reaction is completed, the acid produced and the remaining ester can be separated and collected by a conventional method.
本発明により一般式（１）で示されるエステルを短時間
に効率良く高い光学選択性をもって不斉加水分解出来る
。According to the present invention, the ester represented by the general formula (1) can be asymmetrically hydrolyzed efficiently in a short time with high optical selectivity.
以下、本発明を実施例を挙げて説明するが、本発明はこ
れらの実施例に限定されるものではない。Hereinafter, the present invention will be explained with reference to Examples, but the present invention is not limited to these Examples.
実施例１
φ２１ｍｍの試験管に、リパーゼＣＢ （１７ｍｇ、３
０００）を取り、ｐｏ ８．０の０．２５Ｍリン酸緩衝
液（５ｍＺ）を加え、更にα−クロロプロピオンａ−ｎ
−ブチルエステル（１６５ｍｇ、 １ ｍｍｏｌ）を加
えて、３０°Ｃで４．５時間振盪した。ＩＮ ＮａＯＨ
でｐＨを８にしてから、酢酸エチル（４５ｍりで抽出し
た。有機層を脱水（無水Ｍｇ５Ｏ４）、次いで３０〜４
０℃で濃縮して、〔α：Ｌ”＝ １２．１ （Ｃ＝
１． ＣＨＣｌ３） （９６，８％ｅ、ｅ、）を有する
（Ｓ）−α〜クロロプロピオン酸−ｎ−ブチルエステル
（４９ｍｇ、収率３０％）を得た。水層を６Ｎ ＨＣＩ
でｐＨを１にしてから、酢酸エチルで抽出し、脱水、濃
縮して＠） Ｉｌｌ”−＋５．８４（Ｃ＝ １ 、 Ｃ
ＨＣｌ３） （４０％ｅ、ｅ、）を有する（Ｒ）４−ク
ロロプロピオン酸（７５ｍｇ、収率７０％）を得た。Example 1 Lipase CB (17 mg, 3
000), add 0.25M phosphate buffer (5mZ) with po 8.0, and add α-chloropropion a-n
-butyl ester (165 mg, 1 mmol) was added and shaken at 30°C for 4.5 hours. IN NaOH
The pH was brought to 8 with
Concentrate at 0°C to obtain [α:L”= 12.1 (C=
1. (S)-α-chloropropionic acid-n-butyl ester (49 mg, 30% yield) with CHCl3) (96.8% e,e,) was obtained. Water layer with 6N HCI
After adjusting the pH to 1 with ethyl acetate, it was extracted with ethyl acetate, dehydrated and concentrated.
(R)4-chloropropionic acid (75 mg, yield 70%) with HCl3) (40% e,e,) was obtained.
実施例２
５００−のひだ付三角フラスコに、リパーゼＣＥ（０，
１７ｇ、 ３００００）を取り、ｐＨ８，０の０．２５
Ｍ リン酸緩衝液（５０ｍＺ）を加え、更にα−クロロ
プロピオン酸−ｎ−オクチルエステル（２，２０ｇ、
ＩＯｍｍｏｌ）を加えて、３０℃で６時間攪拌した。Ｉ
Ｎ ＮａＯＨでｐＨを８にしてから、酢酸エチル（４５
０ｍＺ）で抽出した。有機層を脱水（無水Ｍｇ５ｏ４．
）、次いで３０〜４０℃で濃縮して〔α） ｎ”−５，
５０（Ｃ＝３５．ＣｌＩＣ１３）を有する（Ｓ）−α−
クロロプロピオン酸−ｎ−オクチルエステル（１，１９
ｇ、収率５４％）を得た。Example 2 Lipase CE (0,
Take 17g, 30000) and add 0.25 to pH 8.0.
M phosphate buffer (50mZ) was added, and α-chloropropionic acid-n-octyl ester (2.20g,
IO mmol) was added thereto, and the mixture was stirred at 30°C for 6 hours. I
The pH was brought to 8 with N NaOH, then ethyl acetate (45
0 mZ). The organic layer was dehydrated (anhydrous Mg5o4.
), then concentrated at 30-40°C to give [α) n”-5,
(S)-α- with 50 (C=35.ClIC13)
Chloropropionic acid-n-octyl ester (1,19
g, yield 54%).
水層を６Ｎ ＨＣＩでｐＨを１にしてから、酢酸エチル
で抽出し、脱水、濃縮して〔α）ｏ”−＋７．４２（Ｃ
＝１゜ＣＨｚＣｌｚ） （５０，８％ｅ、ｅ、）を有す
る（Ｒ）４−クロロプロピオン酸（０，４８ｇ 、収率
４４％）を得た。The aqueous layer was adjusted to pH 1 with 6N HCl, extracted with ethyl acetate, dehydrated, and concentrated to [α)o”−+7.42(C
(R) 4-chloropropionic acid (0.48 g, yield 44%) with 1°CHzClz) (50.8% e,e,) was obtained.
Claims (1)
式（ I ） ▲数式、化学式、表等があります▼・・・（Ｉ） 〔Ｒ；炭素数が１から２０の飽和または不飽和のアルキ
ル基 Ｒ’；直鎖あるいは分岐のアルキル基、アラルキル基又
はシクロヘキシル基 Ｘ；ハロゲン原子〕 で表されるα−ハロカルボン酸エステルのラセミ体を不
斉加水分解した後、生成する酸と残存するエステルとを
分離・採取することを特徴とする光学活性化合物の製造
法。[Claims] Using an enzyme derived from a microorganism belonging to the genus Humicola, the general formula (I) ▲Mathematical formulas, chemical formulas, tables, etc. are available▼...(I) [R; saturated carbon number of 1 to 20 or unsaturated alkyl group R'; linear or branched alkyl group, aralkyl group, or cyclohexyl group A method for producing an optically active compound, which comprises separating and collecting the remaining ester and the remaining ester.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP4790386A JPS62205797A (en) | 1986-03-05 | 1986-03-05 | Production of optically active compound |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP4790386A JPS62205797A (en) | 1986-03-05 | 1986-03-05 | Production of optically active compound |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| JPS62205797A true JPS62205797A (en) | 1987-09-10 |
Family
ID=12788350
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP4790386A Pending JPS62205797A (en) | 1986-03-05 | 1986-03-05 | Production of optically active compound |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPS62205797A (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5753495A (en) * | 1993-08-23 | 1998-05-19 | Basf Aktiengesellschaft | Process for the preparation of (L)-2-chloropropionic acid and its salts using lipase from pseudomonas |
| KR100846676B1 (en) | 2006-04-27 | 2008-07-16 | 엔자이텍 주식회사 | The method of making optically active 2-halo-2-n-substituted phenylacetic acid esters and 2-halo-2-n-substituted phenylacetic acids by enzymatic method |
-
1986
- 1986-03-05 JP JP4790386A patent/JPS62205797A/en active Pending
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5753495A (en) * | 1993-08-23 | 1998-05-19 | Basf Aktiengesellschaft | Process for the preparation of (L)-2-chloropropionic acid and its salts using lipase from pseudomonas |
| KR100846676B1 (en) | 2006-04-27 | 2008-07-16 | 엔자이텍 주식회사 | The method of making optically active 2-halo-2-n-substituted phenylacetic acid esters and 2-halo-2-n-substituted phenylacetic acids by enzymatic method |
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