JPH0160239B2 - - Google Patents

Info

Publication number
JPH0160239B2
JPH0160239B2 JP17029580A JP17029580A JPH0160239B2 JP H0160239 B2 JPH0160239 B2 JP H0160239B2 JP 17029580 A JP17029580 A JP 17029580A JP 17029580 A JP17029580 A JP 17029580A JP H0160239 B2 JPH0160239 B2 JP H0160239B2
Authority
JP
Japan
Prior art keywords
acid
bromopropionate
ethyl
added
hours
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.)
Expired
Application number
JP17029580A
Other languages
Japanese (ja)
Other versions
JPS5794295A (en
Inventor
Shinobu Iriuchijima
Natsuko Kojima
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Sagami Chemical Research Institute
Original Assignee
Sagami Chemical Research Institute
Priority date (The priority date 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 date listed.)
Filing date
Publication date
Application filed by Sagami Chemical Research Institute filed Critical Sagami Chemical Research Institute
Priority to JP17029580A priority Critical patent/JPS5794295A/en
Publication of JPS5794295A publication Critical patent/JPS5794295A/en
Publication of JPH0160239B2 publication Critical patent/JPH0160239B2/ja
Granted legal-status Critical Current

Links

Description

【発明の詳細な説明】[Detailed description of the invention]

本発明は一般式 (式中、Rはアルキル基であり、R′はアルキル
基、置換アルキル基又はアリール基であり、Xは
ハロゲン原子である。)で表わされるエステルを、
酵素又は微生物を用いて不斉水解した後、残存し
たエステルと生成した酸を分離、取得することを
特徴とする光学活性のエステル及び/又は酸の製
造法に関する。 前記一般式()で表わされるエステル又はそ
の酸の光学活性体は、光学活性基幹物質であり、
種々の有用な光学活性化合物の合成に用いること
ができる極めて重要な化合物である。例えば、L
−2−ブロモプロピオン酸メチルは、除草活性の
強いD型の除草剤の合成に利用でき(特開昭54−
125682号参照。)、(R)−(+)−2−ブロモプロピ
オン酸(L型)は、抗オーキシン活性の強い
(S)−(−)−2−(ベンズイミダゾリル−2−チ
オ)プロピオン酸(D型)の合成に用いられる
〔Chem.Abstr.、88、105226y(1978)、参照。〕。
又、L−2−クロロプロピオン酸は免疫アジユバ
ント活性の強いムラミルペプチドの合成に用いら
れる(Tetrahedron Lett.、1978、4407参照。)。 従来、前記一般式()で表わされるエステル
又はその酸の光学活性体は、(1)光学活性の乳酸を
エステル化してから三臭化リン又は三塩化リンを
反応させる方法〔J.Am.Chem.Soc.、95、7908
(1973)、参照。〕、(2)(S)−アラニンに酸性条件
下臭化カリウムと亜硝酸ナトリウムを反応させる
方法(J.C.S.Perkin I、1976、2229参照。)、又
は(3)α−ハロカルボン酸を光学活性有機塩基を用
いて光学分割する方法〔J.Am.Chem.Soc.、92、
5519(1970)及び引用文献参照。〕等によつて合成
された。しかしながら、(1)の方法は高価な天然物
を用い、しかも三ハロゲン化リンの如き刺激臭の
強い試薬を用いなければならず、又、(2)の方法は
強酸性条件下0℃で、(S)−アラニンに対し6当
量の臭化カリウムと2.7当量の亜硝酸ナトリウム
を用いるジアゾ化反応であり、(3)の方法はシンコ
ニンの如き高価な有機塩基を用いて再結晶を繰り
返さなければならず、いずれも工業的製造法とは
なり難い。 本発明者等は、前記一般式()で表わされる
エステル又はその酸の光学活性体が、広く医薬、
農薬等の合成に用いられる有用な化合物であるこ
とに鑑み、その工業的製造法について鋭意研究を
重ねた結果、従来の方法とは全く異なる、操作の
極めて容易な本発明の方法を見い出したものであ
る。 本方法に基質として用いるエステルの酸部分と
しては、2−クロロプロピオン酸、2−ブロモプ
ロピオン酸の如き2−ハロプロピオン酸、2−ク
ロロ酪酸、2−ブロモ酪酸の如き2−ハロ酪酸、
2−クロロ吉草酸、2−ブロモ吉草酸の如き2−
ハロ吉草酸を例示することができる。また、エス
テルのアルコール部分としては、メタノール、エ
タノール、プロパノール、ブタノール、イソブタ
ノール、ベンジルアルコール、フエノール等を挙
げることができる。 本方法に用いることのできる酵素は、エステル
を不斉水解する能力を有する酵素であり、例え
ば、表1に示した酵素を挙げることができる。ま
た、本方法に用いることのできる微生物は、例え
ば、表1の「起源」の項に示した微生物及びリゾ
プスデレマーIAM6019、バチルスズブチリスバ
ル・ニガーIFO3108を挙げることができる。
The present invention is based on the general formula (In the formula, R is an alkyl group, R' is an alkyl group, substituted alkyl group, or aryl group, and X is a halogen atom.)
The present invention relates to a method for producing optically active esters and/or acids, which comprises separating and obtaining residual esters and generated acids after asymmetric hydrolysis using enzymes or microorganisms. The ester represented by the general formula () or an optically active form of its acid is an optically active base substance,
It is an extremely important compound that can be used in the synthesis of various useful optically active compounds. For example, L
-Methyl 2-bromopropionate can be used to synthesize D-type herbicides with strong herbicidal activity (Japanese Patent Application Laid-open No. 1983-1996-
See No. 125682. ), (R)-(+)-2-bromopropionic acid (L type) is (S)-(-)-2-(benzimidazolyl-2-thio)propionic acid (D type), which has strong anti-auxin activity. [See Chem.Abstr., 88, 105226y (1978). ].
Furthermore, L-2-chloropropionic acid is used for the synthesis of muramyl peptide, which has strong immune adjuvant activity (see Tetrahedron Lett., 1978, 4407). Conventionally, the ester represented by the general formula () or the optically active form of its acid has been prepared by (1) a method of esterifying optically active lactic acid and then reacting it with phosphorus tribromide or phosphorus trichloride [J.Am.Chem .Soc., 95, 7908
(1973), see. ], (2) A method in which (S)-alanine is reacted with potassium bromide and sodium nitrite under acidic conditions (see JCS Perkin I, 1976, 2229), or (3) a method in which α-halocarboxylic acid is reacted with an optically active organic base. [J.Am.Chem.Soc., 92,
5519 (1970) and cited references. ] and others. However, method (1) requires the use of expensive natural products and reagents with strong irritating odors such as phosphorus trihalide, and method (2) requires the use of expensive natural products and reagents with a strong odor such as phosphorus trihalide. This is a diazotization reaction using 6 equivalents of potassium bromide and 2.7 equivalents of sodium nitrite for (S)-alanine, and method (3) requires repeated recrystallization using an expensive organic base such as cinchonine. Neither of these methods can be used as an industrial manufacturing method. The present inventors believe that the ester represented by the general formula () or the optically active form of its acid can be widely used in pharmaceuticals,
In view of the fact that it is a useful compound used in the synthesis of agricultural chemicals, etc., as a result of intensive research on its industrial production method, we have discovered the method of the present invention, which is completely different from conventional methods and is extremely easy to operate. It is. The acid moiety of the ester used as a substrate in this method includes 2-halopropionic acids such as 2-chloropropionic acid and 2-bromopropionic acid, 2-halobutyric acids such as 2-chlorobutyric acid and 2-bromobutyric acid,
2-chlorovaleric acid, 2-bromovaleric acid, etc.
Halovaleric acid can be exemplified. Further, examples of the alcohol moiety of the ester include methanol, ethanol, propanol, butanol, isobutanol, benzyl alcohol, and phenol. Enzymes that can be used in this method are enzymes that have the ability to asymmetrically hydrolyze esters, and include, for example, the enzymes shown in Table 1. In addition, examples of microorganisms that can be used in this method include the microorganisms shown in the "Origin" section of Table 1, Rhizopus deremer IAM6019, and Bacillus subtilisbar niger IFO3108.

【表】【table】

【表】 これら酵素又は微生物は、精製酵素、粗酵素、
酵素含有物、微生物培養液、培養物、菌体、培養
濾液及びそれらを処理した物など種々の形体で必
要に応じて用いることができる。更に、酵素と微
生物を組み合わせて用いることもできる。 本発明の方法の実施に当つては、使用する酵素
又は微生物にもよるが、通常、緩衝液の使用が好
ましく、リン酸ナトリウム、リン酸カリウムの如
き無機酸塩の緩衝液、クエン酸ナトリウムの如き
有機酸塩の緩衝液を好適に使用することができ
る。初発PHは7〜8が好適に使用でき、反応の間
4〜7.5に保たれることが望ましい。濃度は、緩
衝液の種類にもよるが、0.05〜1Mが使用でき、
0.1〜0.5Mが好適に使用することができる。 水解によつて生成する酸によつて極度な酸性に
ならない様に緩衝液を用い、不斉水解能を有する
酵素又は微生物及び基質を加え、数時間乃至数日
間、撹拌又は振とうを行なう。反応は10〜50℃で
実施できるが、低温では反応が遅くなり、高温で
は酵素の失活及び不斉選択性の低下が見られるこ
とがあるので、20〜35℃が望ましい。反応終了
後、通常の方法によつてエステルと酸を分離す
る。 以下、実施例により更に具体的に説明する。 実施例 1 500mlの三角フラスコに、リパーゼMY(30mg)
を取り、PH7.5の0.2Mリン酸ナトリウム緩衝液
(以下の実施例において特記しない限り本緩衝液
を用いた。尚、その際の記載は単に「緩衝液」と
した。)(500ml)を加え、更に2−ブロモプロピ
オン酸エチル(2,2−ジブロモプロピオン酸エ
チルと思われる不純物を少量含む。)(21.0g、
116mmol)を加えて、30℃で17時間撹拌した。
1M炭酸水素ナトリウム水溶液(7ml)を加えて
から、酢酸エチル(200ml)で抽出した。有機層
を脱水(無水硫酸ナトリウム)、次いで30〜40℃
で濃縮し、更にbp60〜65℃/20mmHgで蒸留し
て、〔α〕25 D−26.4゜(c=1.09、CH2Cl2)(92%e.e
.)
〔文献値:〔α〕D28.8゜(CH2Cl2)、J.Am.Chem.
Soc.、95、7908(1973)参照〕及び〔α〕25 D−29.4゜
(neat)(92%e.e.)〔文献値:〔α〕D31.9゜(neat
)、
同上文献参照。〕を有する(S)−2−ブロモプロ
ピオン酸エチル(4.55g、22%)を得た。このも
ののNMRスペクトルを測定したところ、使用し
た試薬中に混入している2,2−ジブロモプロピ
オン酸エチルと思われる化合物〔NMR(CDCl3
δ:1.35(t、J=7Hz)、2.66(s)〕が約10%
(重量)入つていることがわかつた。このことを
考慮に入れると、得られた(S)−2−ブロモプ
ロピオン酸エチルは光学的にほとんど純粋である
と考えられる。 実施例 2 125mlの三角フラスコに、リゾプスデレマーリ
パーゼ(41mg)を取り、緩衝液(100ml)を加え、
更に2−ブロモプロピオン酸エチル(4.2g、
23.2mmol)を加えて、25℃で23時間撹拌した。
6N硫酸(2ml)で酸性にしてから、酢酸エチル
(40ml)で抽出した。有機層を1M炭酸水素ナトリ
ウム(20ml)と振りまぜた。有機層を脱水した
後、濃縮し、〔α〕25 D−16.3゜(c=1.26、CH2Cl2
(57%e.e.)及び〔α〕25 D−20.2゜(neat)(63%e.e
.)
を有する(S)−2−ブロモプロピオン酸エチル
(1.46g、35%)を得た。水層を6N硫酸で酸性に
してから、酢酸エチルで抽出し、脱水、濃縮し
て、〔α〕25 D+17.9゜(c=1.11、CH2Cl2)を有する
(R)−2−ブロモプロピオン酸(1.79g、50%)
を得た。バス温(減圧度)70〜85℃(1mmHg)
で蒸留して、〔α〕25 D+19.5゜(c=1.21、CH2Cl2
及び〔α〕25 D+16.9゜(neat)(53%e.e.)〔文献値

〔α〕20 D−27.1゜(neat)、J.C.S.Perkin I、1976、
2229、参照。〕を有するもの(795mg22%)を得
た。 実施例 3 試験管(25×200)に、リゾブスリパーゼT(12
mg)を取り、緩衝液(PH7)(15ml)を加え、次
いで2−ブロモプロピオン酸エチル(0.70g、
3.87mmol)を加えて、30℃で16時間試験管振と
う機で振とうした。6N硫酸(0.3ml)を加え、酢
酸エチル(7ml)で抽出した。有機層を1M炭酸
水素ナトリウム(4ml)で洗い、脱水、濃縮し
て、〔α〕25 D−20.9゜(c=1.19、CH2Cl2)(73%e.e
.)
を有する(S)−2−ブロモプロピオン酸エチル
(179mg、25.6%)を得た。 実施例 4〜7 表2に特記した以外は、実施例3と同様に行な
い、表2の結果を得た。
[Table] These enzymes or microorganisms are purified enzymes, crude enzymes,
It can be used in various forms as needed, such as enzyme-containing materials, microbial culture solutions, cultures, bacterial cells, culture filtrates, and processed products. Furthermore, enzymes and microorganisms can be used in combination. When carrying out the method of the present invention, it is usually preferable to use a buffer solution, although it depends on the enzyme or microorganism used. Buffers of organic acid salts such as the following can be suitably used. An initial pH of 7 to 8 can be suitably used, and it is desirable to maintain it at 4 to 7.5 during the reaction. The concentration depends on the type of buffer solution, but 0.05 to 1M can be used.
0.1-0.5M can be suitably used. A buffer solution is used to prevent the mixture from becoming extremely acidic due to the acid generated by water decomposition, and an enzyme or microorganism capable of asymmetric water decomposition and a substrate are added, followed by stirring or shaking for several hours to several days. The reaction can be carried out at a temperature of 10 to 50°C, but a temperature of 20 to 35°C is preferable, since the reaction slows down at low temperatures and deactivation of the enzyme and a decrease in chiral selectivity may occur at high temperatures. After the reaction is complete, the ester and acid are separated by a conventional method. Hereinafter, a more specific explanation will be given with reference to Examples. Example 1 Lipase MY (30mg) in a 500ml Erlenmeyer flask
and 500 ml of 0.2M sodium phosphate buffer (PH7.5) (this buffer was used in the following examples unless otherwise specified. In such cases, the description was simply "buffer"). In addition, ethyl 2-bromopropionate (contains a small amount of impurity thought to be ethyl 2,2-dibromopropionate) (21.0 g,
116 mmol) was added thereto, and the mixture was stirred at 30°C for 17 hours.
A 1M aqueous sodium bicarbonate solution (7 ml) was added, followed by extraction with ethyl acetate (200 ml). The organic layer is dehydrated (anhydrous sodium sulfate) and then heated to 30-40°C.
Concentrate at _ _
.)
[Literature value: [α] D 28.8° (CH 2 Cl 2 ), J.Am.Chem.
Soc., 95, 7908 (1973)] and [α] 25 D −29.4° (neat) (92% ee) [Literature value: [α] D 31.9° (neat
),
See the same reference. ] (S)-2-bromopropionate (4.55 g, 22%) was obtained. When the NMR spectrum of this product was measured, it was found that a compound believed to be ethyl 2,2-dibromopropionate [NMR (CDCl 3 )] was found mixed in the reagent used.
δ: 1.35 (t, J = 7Hz), 2.66 (s)] is approximately 10%
(Weight) I found out that it was included. Taking this into consideration, the obtained ethyl (S)-2-bromopropionate is considered to be almost optically pure. Example 2 Take Rhizopus deremer lipase (41 mg) into a 125 ml Erlenmeyer flask, add buffer solution (100 ml),
Furthermore, ethyl 2-bromopropionate (4.2 g,
23.2 mmol) was added thereto, and the mixture was stirred at 25°C for 23 hours.
The mixture was acidified with 6N sulfuric acid (2 ml) and extracted with ethyl acetate (40 ml). The organic layer was mixed with 1M sodium bicarbonate (20ml). After dehydrating the organic layer, it was concentrated and [α] 25 D −16.3° (c=1.26, CH 2 Cl 2 )
(57%ee) and [α] 25 D −20.2゜(neat) (63%ee
.)
Ethyl (S)-2-bromopropionate (1.46 g, 35%) was obtained. The aqueous layer was acidified with 6N sulfuric acid, extracted with ethyl acetate, dried and concentrated to give (R)-2- with [α] 25 D +17.9° (c=1.11, CH 2 Cl 2 ). Bromopropionic acid (1.79g, 50%)
I got it. Bath temperature (degree of vacuum) 70-85℃ (1mmHg)
Distill with [α] 25 D +19.5° (c=1.21, CH 2 Cl 2 )
and [α] 25 D +16.9° (neat) (53%ee) [Literature value:
[α] 20 D −27.1゜(neat), JCSPerkin I, 1976,
2229, see. ] (795mg22%) was obtained. Example 3 Rhizobus lipase T (12
mg), add buffer solution (PH7) (15 ml), and then add ethyl 2-bromopropionate (0.70 g,
3.87 mmol) was added thereto, and the mixture was shaken in a test tube shaker at 30°C for 16 hours. 6N sulfuric acid (0.3ml) was added, and the mixture was extracted with ethyl acetate (7ml). The organic layer was washed with 1M sodium bicarbonate (4 ml), dried, and concentrated to give [α] 25 D -20.9° (c = 1.19, CH 2 Cl 2 ) (73% ee).
.)
Ethyl (S)-2-bromopropionate (179 mg, 25.6%) was obtained. Examples 4 to 7 Except as noted in Table 2, the same procedure as in Example 3 was carried out, and the results shown in Table 2 were obtained.

【表】 実施例 8 リゾプスデレマーIAM6019を、培地Y*(100
ml)の入つた500mlの三角フラスコ1本を用い、
30℃の48時間振とう培養した。培養物を濾過して
生乾きの菌体(3g)を得た。このもの(0.3g)
を50mlの三角フラスコに取り、緩衝液(30ml)を
加え、更に2−ブロモプロピオン酸エチル(1.40
g、7.73mmol)を加えて、29℃で19時間撹拌し
た。6N硫酸(1ml)及び酢酸エチルを加え振と
うした後、濾過して菌体を除いた。濾液より分け
取つた有機層を、1M炭酸水素ナトリウム(8ml)
で洗い、脱水、濃縮して、〔α〕25 D−15.8゜(c=
1.18、CH2Cl2)(55%e.e.)を有する(S)−2−
ブロモプロピオン酸エチル(0.46g、33%)を得
た。 *培地Y:グルコース1%、酵母エキス0.3%、
麦芽エキス0.3%、ペプトン0.5%、PH5〜6。 実施例 9 実施例8と同様に行なつて得られたリゾプスデ
レマーIAM6019の培養液(100ml)に、緩衝液
(300ml)を加え、更に2−ブロモプロピオン酸エ
チル(14.0g、77.3mmol)を加えて、室温で10
時間撹拌した。1M炭酸水素ナトリウム(20ml)
を改えて酢酸エチルで抽出した。有機層を脱水、
濃縮、バス温(減圧度)70〜85℃(16mmHg)で
蒸留して、〔α〕25 D−21.2゜(c=1.09、CH2Cl2)(7
4
%e.e.)及び〔α〕25 D−23.5゜(neat)(74%e.e.)

有する(S)−2−ブロモプロピオン酸エチル
(3.30g、24%)を得た。 水層を6N硫酸で酸性にして酢酸エチルで抽出
した。抽出液を脱水、濃縮、bp103〜6℃/16mm
Hgで蒸留して、〔α〕25 D+16.8゜(c=1.19、
CH2Cl2)及び〔α〕25 D+14.4゜(neat)(53%e.e.)
を有する(R)−2−ブロモプロピオン酸(3.9
g、33%)を得た。 実施例 10 リパーゼMAP10(196mg)を取り、緩衝液(50
ml)を加え、更に2−ブロモプロピオン酸エチル
(2.10g、11.6mmol)を加えて、30℃で8.5時間
撹拌した。1M炭酸水素ナトリウム(2ml)を加
えて酢酸エチル(25ml)で抽出した。有機層を脱
水、濃縮して、〔α〕25 D−16.1゜(c=1.22、CH2Cl2

(56%e.e.)を有する(S)−2−ブロモプロピオ
ン酸エチル(0.63g、30%)を得た。 実施例 11 表3に特記した以外は、実施例10と同様に行
い、表3の結果を得た。
[Table] Example 8 Rhizopus deremer IAM6019 was grown in medium Y * (100
ml) using one 500ml Erlenmeyer flask containing
Shaking culture was performed at 30°C for 48 hours. The culture was filtered to obtain half-dried bacterial cells (3 g). This thing (0.3g)
into a 50 ml Erlenmeyer flask, add buffer (30 ml), and add ethyl 2-bromopropionate (1.40 ml).
g, 7.73 mmol) and stirred at 29°C for 19 hours. After adding 6N sulfuric acid (1 ml) and ethyl acetate and shaking, the mixture was filtered to remove bacterial cells. The organic layer separated from the filtrate was added to 1M sodium hydrogen carbonate (8 ml).
Wash, dehydrate, and concentrate to obtain [α] 25 D −15.8° (c=
(S)-2- with 1.18, CH 2 Cl 2 ) (55% ee)
Ethyl bromopropionate (0.46 g, 33%) was obtained. * Medium Y: glucose 1%, yeast extract 0.3%,
Malt extract 0.3%, peptone 0.5%, PH5-6. Example 9 A buffer solution (300 ml) was added to a culture solution (100 ml) of Rhizopus deremer IAM6019 obtained in the same manner as in Example 8, and ethyl 2-bromopropionate (14.0 g, 77.3 mmol) was added. , 10 at room temperature
Stir for hours. 1M sodium bicarbonate (20ml)
was extracted again with ethyl acetate. Dehydrate the organic layer;
Concentrate and distill at a bath temperature (degree of vacuum) of 70 to 85°C (16 mmHg) to obtain [α] 25 D -21.2° (c = 1.09, CH 2 Cl 2 ) (7
Four
%ee) and [α] 25 D −23.5° (neat) (74%ee)
Ethyl (S)-2-bromopropionate (3.30 g, 24%) was obtained. The aqueous layer was acidified with 6N sulfuric acid and extracted with ethyl acetate. Dehydrate the extract, concentrate, bp103~6℃/16mm
Distilled with Hg, [α] 25 D +16.8° (c=1.19,
CH 2 Cl 2 ) and [α] 25 D +14.4° (neat) (53%ee)
(R)-2-bromopropionic acid (3.9
g, 33%) was obtained. Example 10 Take lipase MAP10 (196 mg) and add buffer solution (50
ml), ethyl 2-bromopropionate (2.10 g, 11.6 mmol) was added, and the mixture was stirred at 30°C for 8.5 hours. 1M sodium hydrogen carbonate (2 ml) was added and the mixture was extracted with ethyl acetate (25 ml). The organic layer was dehydrated and concentrated to give [α] 25 D −16.1° (c=1.22, CH 2 Cl 2
)
Ethyl (S)-2-bromopropionate (0.63 g, 30%) with (56% ee) was obtained. Example 11 Except as noted in Table 3, the same procedure as Example 10 was carried out, and the results shown in Table 3 were obtained.

【表】 実施例 12 バチルスズブチリスバル、ニガーIFO3108を、
培地B*(10ml)の入つた試験管1本を使い、30℃
で24時間振とう培養した。 この培養液を50mlの三角フラスコに移し、緩衝
液(30ml)に加え、更に2−ブロモプロピオン酸
エチル(2.10g、11.6mmol)を加えて、30℃で
18時間撹拌した。6N硫酸(1ml)を加え、酢酸
エチルで抽出した。有機酸を1M炭酸水素ナトリ
ウム(12ml)で洗い、脱水、濃縮して、〔α〕25 D
17.5゜(c=1.02、CH2Cl2)(61%e.e.)を有する
(R)−2−ブロモプロピオン酸エチル(490ml、
23%)を得た。 *培地B:粉末ブイヨン2%、酵母エキス0.5%、
PH7。 実施例 13 100mlの三角フラスコに、リパーゼMY(2mg)
を取り、緩衝液(100ml)を加え、更に2−クロ
ロプロピオン酸メチル(3.24g、26.4mmol)を
加えて、30℃で18時間撹拌した。1M炭酸水素ナ
トリウム(5ml)を加えて塩化メチレン(40ml)
で抽出した後、有機層を脱水し、溶媒を常圧蒸留
で除去した後、バス温135〜150℃で蒸留し、〔α〕
25 D−10.8℃(c=1.07、CH2Cl2)及び〔α〕25 D
12.4゜(neat)(46%e.e.)〔文献値:〔α〕D−26.83

(neat)、J.Chem.Soc.Trans.、67、914(1895)参
照。〕を有する(S)−2−クロロプロピオン酸メ
チル(0.725g、22%)を得た。 実施例 14 リパーゼMY(13.5mg)を取り、緩衝液(10ml)
を加え、更に2−ブロモプロピオン酸エチル
(2.10g、11.6mmol)を加えて30℃で7時間撹拌
した。この間PH4〜7に保たれる様に、1M炭酸
水素ナトリウム(6ml)を適宜滴下した。酢酸エ
チル(10ml)で抽出し、脱水、濃縮して、〔α〕25 D
−21.2゜(c=1.27、CH2Cl2)(74%e.e.)を有する
(S)−2−ブロモプロピオン酸エチル(0.665g、
32%)を得た。 実施例 32 リパーゼMY(200mg)をPH7.5の0.5Mリン酸ナ
トリウム緩衝液(22ml)に溶かし、α−ブロモイ
ソ吉草酸メチル(2.04g)を加え、30℃で42時間
撹拌した。最終PH7であつた。塩化メチレンで抽
出し、脱水、濃縮して、〔α〕25 D+18.1゜(neat)を
有するα−ブロモイソ吉草酸メチル(0.80g、40
%)を得た。本生成物の光学純度はシフト試薬
Eu(HFC)3を添加したNMRスペクトルによつて
約50%e.e.と決定された。 実施例 33 リパーゼMY(15mg)をPH7.5の0.8Mリン酸ナト
リウム緩衝液(55ml)に溶かし、2−ブロモプロ
ピオン酸エチル(9.24g)を加え、30℃で24時間
撹拌した。反応後、塩化メチレンで抽出し、脱
水、濃縮して、〔α〕25 D−30.0゜(neat)(94%e.e.

を有する(S)−2−ブロモプロピオン酸エチル
(2.66g、29%)を得た。 実施例 34 リパーゼMY(25mg)を緩衝液(100ml)に溶か
し、2−ブロモプロピオン酸メチル(4.5g)を
加え、30℃で22時間撹拌した。最終PH4であつた
ので、1M炭酸水素ナトリウムでPH7とし塩化メ
チレンで抽出した。抽出液を脱水、濃縮して、
〔α〕25 D−43.1゜(neat)を有する(S)−2−ブロ

プロピオン酸エチル(1.12g、25%)を得た。光
学純度はシフト試薬Eu(HFC)3を添加したNMR
スペクトルより約90%e.e.と決定された。 実施例 35 リパーゼMY(5mg)をPH7.5の0.8Mリン酸ナト
リウム緩衝液(20ml)に溶かし、2−ブロモプロ
ピオン酸イソブチル(2.8g)を加え、30℃で
25.5時間撹拌した。反応後のPH6であつたので、
1M炭酸水素ナトリウムでPH7とし、塩化メチレ
ンで抽出した。抽出液を脱水、濃縮、蒸留して、
〔α〕25 D−18.1゜(neat)を有する(S)−2−ブロ

プロピオン酸イソブチル(0.63g、22.5%)を得
た。光学純度はEu(HFC)3を添加したNMRスペ
クトルにより、約90%e.e.と決定された。 実施例 36 リパーゼMY(5mg)をPH7.5の0.8Mリン酸ナト
リウム緩衝液(18ml)に溶かし、2−ブロモプロ
ピオン酸ブチル(2.8g)を加え、30℃で20時間
撹拌した。1M炭酸水素ナトリウムでPH7とし、
塩化メチレンで抽出した。抽出液を脱水、濃縮、
蒸留して、〔α〕25 D−21゜(neat)を有する(S)−
2−ブロモプロピオン酸ブチル(0.75g、26.6
%)を得た。光学純度はEu(HFC)3を添加した
NMRスペクトルより、約90%e.e.と決定された。
[Table] Example 12 Bacillus subtilis bacillus, niger IFO3108,
Using one test tube containing medium B * (10 ml), heat at 30°C.
Cultured with shaking for 24 hours. Transfer this culture solution to a 50 ml Erlenmeyer flask, add buffer solution (30 ml) and further add ethyl 2-bromopropionate (2.10 g, 11.6 mmol), and heat at 30°C.
Stirred for 18 hours. 6N sulfuric acid (1 ml) was added, and the mixture was extracted with ethyl acetate. Wash the organic acid with 1M sodium bicarbonate (12 ml), dehydrate, and concentrate to obtain [α] 25 D +
Ethyl (R) -2 - bromopropionate (490 ml,
23%). * Medium B: powdered bouillon 2%, yeast extract 0.5%,
PH7. Example 13 Lipase MY (2 mg) in a 100 ml Erlenmeyer flask
A buffer solution (100 ml) was added thereto, followed by methyl 2-chloropropionate (3.24 g, 26.4 mmol), and the mixture was stirred at 30°C for 18 hours. Add 1M sodium bicarbonate (5 ml) and methylene chloride (40 ml).
After extraction, the organic layer was dehydrated, the solvent was removed by atmospheric distillation, and then distilled at a bath temperature of 135-150℃, [α]
25 D −10.8°C (c=1.07, CH 2 Cl 2 ) and [α] 25 D
12.4゜(neat) (46%ee) [Literature value: [α] D −26.83
See ゜(neat), J.Chem.Soc.Trans., 67 , 914 (1895). ] methyl (S)-2-chloropropionate (0.725 g, 22%) was obtained. Example 14 Take lipase MY (13.5 mg) and add buffer solution (10 ml)
was added, and further ethyl 2-bromopropionate (2.10 g, 11.6 mmol) was added, followed by stirring at 30°C for 7 hours. During this time, 1M sodium hydrogen carbonate (6 ml) was appropriately added dropwise to maintain the pH at 4 to 7. Extract with ethyl acetate (10 ml), dehydrate, and concentrate to give [α] 25 D
Ethyl (S)-2-bromopropionate (0.665 g, with -21.2° (c=1.27, CH 2 Cl 2 ) (74% ee)
32%). Example 32 Lipase MY (200 mg) was dissolved in 0.5M sodium phosphate buffer (22 ml) at pH 7.5, methyl α-bromoisovalerate (2.04 g) was added, and the mixture was stirred at 30°C for 42 hours. The final pH was 7. Extracted with methylene chloride, dried and concentrated to obtain methyl α- bromoisovalerate (0.80 g, 40
%) was obtained. The optical purity of this product is determined by the shift reagent
Approximately 50% ee was determined by NMR spectra with addition of Eu(HFC) 3 . Example 33 Lipase MY (15 mg) was dissolved in 0.8M sodium phosphate buffer (55 ml) at pH 7.5, ethyl 2-bromopropionate (9.24 g) was added, and the mixture was stirred at 30°C for 24 hours. After the reaction, it was extracted with methylene chloride, dehydrated, and concentrated to obtain [α] 25 D −30.0° (neat) (94% ee
)
Ethyl (S)-2-bromopropionate (2.66 g, 29%) was obtained. Example 34 Lipase MY (25 mg) was dissolved in a buffer solution (100 ml), methyl 2-bromopropionate (4.5 g) was added, and the mixture was stirred at 30°C for 22 hours. Since the final pH was 4, the pH was adjusted to 7 with 1M sodium hydrogen carbonate and extracted with methylene chloride. Dehydrate and concentrate the extract,
Ethyl (S)-2-bromopropionate (1.12 g, 25%) having [α] 25 D -43.1° (neat) was obtained. Optical purity is determined by NMR with the addition of shift reagent Eu(HFC) 3
It was determined to be approximately 90% ee from the spectrum. Example 35 Lipase MY (5 mg) was dissolved in 0.8 M sodium phosphate buffer (20 ml) at pH 7.5, wasobutyl 2-bromopropionate (2.8 g) was added, and the mixture was incubated at 30°C.
Stirred for 25.5 hours. Since the pH was 6 after the reaction,
The pH was adjusted to 7 with 1M sodium hydrogen carbonate, and the mixture was extracted with methylene chloride. The extract is dehydrated, concentrated, and distilled.
Isobutyl (S)-2-bromopropionate (0.63 g, 22.5%) having [α] 25 D -18.1° (neat) was obtained. The optical purity was determined to be approximately 90% ee by NMR spectroscopy with the addition of Eu(HFC) 3 . Example 36 Lipase MY (5 mg) was dissolved in 0.8M sodium phosphate buffer (18 ml) at pH 7.5, butyl 2-bromopropionate (2.8 g) was added, and the mixture was stirred at 30°C for 20 hours. Adjust the pH to 7 with 1M sodium bicarbonate,
Extracted with methylene chloride. Dehydrate the extract, concentrate,
Distilled to give (S)- having [α] 25 D −21° (neat)
Butyl 2-bromopropionate (0.75g, 26.6
%) was obtained. Optical purity was added with Eu(HFC) 3
From the NMR spectrum, it was determined to be approximately 90% ee.

Claims (1)

【特許請求の範囲】 1 エステルを不斉水解する能力を有するリゾプ
ス属、ムコル属、カンジタ属、バチルス属の各属
に属する微生物から得られた酵素又はリゾプス
属、ムコル属、カンジタ属、バチルス属の各属に
属する微生物を用いて、一般式 で表わされるエステルを不斉水解した後、残存し
たエステルと生成した酸を分離、取得することを
特徴とする光学活性のエステル及び/又は酸の製
造法(前記一般式において、Rはアルキル基であ
り、R′はアルキル基、置換アルキル基又はアリ
ール基であり、Xはハロゲン原子である。)。
[Scope of Claims] 1. Enzymes obtained from microorganisms belonging to the genera Rhizopus, Mucor, Candida, and Bacillus that have the ability to asymmetrically hydrolyze esters, or enzymes of the genus Rhizopus, Mucor, Candida, and Bacillus. Using microorganisms belonging to each genus, the general formula A method for producing an optically active ester and/or acid, which comprises asymmetrically hydrolyzing an ester represented by the formula, and then separating and obtaining the remaining ester and the generated acid (in the general formula, R is an alkyl group). (R' is an alkyl group, substituted alkyl group or aryl group, and X is a halogen atom).
JP17029580A 1980-12-04 1980-12-04 Preparation of optically active ester and/or acid Granted JPS5794295A (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
JP17029580A JPS5794295A (en) 1980-12-04 1980-12-04 Preparation of optically active ester and/or acid

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
JP17029580A JPS5794295A (en) 1980-12-04 1980-12-04 Preparation of optically active ester and/or acid

Publications (2)

Publication Number Publication Date
JPS5794295A JPS5794295A (en) 1982-06-11
JPH0160239B2 true JPH0160239B2 (en) 1989-12-21

Family

ID=15902308

Family Applications (1)

Application Number Title Priority Date Filing Date
JP17029580A Granted JPS5794295A (en) 1980-12-04 1980-12-04 Preparation of optically active ester and/or acid

Country Status (1)

Country Link
JP (1) JPS5794295A (en)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2524744Y2 (en) * 1991-08-30 1997-02-05 積水化成品工業株式会社 Adhesive label

Families Citing this family (11)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0130752B1 (en) * 1983-07-04 1991-02-27 Mitsubishi Rayon Co., Ltd. Process for preparing optically active carboxylic acids and antipode esters thereof
US4584270A (en) * 1984-01-23 1986-04-22 Wisconsin Alumni Research Foundation Process for preparing optically-active 4-amino-3-hydroxybutyric acid
US4601987A (en) * 1985-02-27 1986-07-22 Massachusetts Institute Of Technology Enzymatic production of optical isomers of 2-halopropionic acids
US4668628A (en) * 1985-04-01 1987-05-26 Stauffer Chemical Company Resolution of racemic mixtures of aliphatic acid esters
DK620486A (en) * 1985-12-20 1987-06-21 Wisconsin Alumni Res Found PROCEDURE FOR THE PREPARATION OF ACETIC ACID DERIVATIVES
US5248601A (en) * 1986-03-14 1993-09-28 Franco Francalanci Process for preparing L(-)-carnitine chloride
US5037747A (en) * 1988-01-26 1991-08-06 Hoffmann-La Roche Inc. Production of benzopyran-2-carboxylic acids and esters by enzymatic hydrolysis
US5061629A (en) * 1988-01-26 1991-10-29 Hoffman-La Roche Inc. Production of 2-hydroxy substituted arylalkanoic acids and esters by enzymatic hydrolysis
US4923810A (en) * 1988-08-24 1990-05-08 Genzyme Corporation Resolution of glycidyl esters to high enantiomeric excess
EP0475255A3 (en) * 1990-09-12 1993-04-14 F. Hoffmann-La Roche Ag Process for the preparation of optically pure (s)-alpha-((tert-butylsulfonyl)methyl)hydro cinnamic acid
DE4328231C1 (en) * 1993-08-23 1994-07-28 Basf Ag Process for the preparation of (L) -2-chloropropionic acid and its salts

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2524744Y2 (en) * 1991-08-30 1997-02-05 積水化成品工業株式会社 Adhesive label

Also Published As

Publication number Publication date
JPS5794295A (en) 1982-06-11

Similar Documents

Publication Publication Date Title
JP2707076B2 (en) Production method of optically active compound
EP0195717A2 (en) Process for the biotechnological preparation of optically active alpha-arylalkanoic acids
JPH0160239B2 (en)
JPH047195B2 (en)
FR2497230A1 (en) PROCESS FOR THE PREPARATION OF OPTICALLY ACTIVE MONOALKYL ESTERS OF B- (S) -AMINOGLUTARIC ACID
JPH06256278A (en) Optically active alpha-carbamoylalkanoic acid derivative and its production
JP2698627B2 (en) Method for producing optically active amines and derivatives thereof
JP2542833B2 (en) Process for producing optically active alcohols and esters
JP3732535B2 (en) Process for producing optically active α-methylalkanedicarboxylic acid-ω-monoester and its enantiomer diester
JP2579766B2 (en) Optically active biphenyl derivative and method for producing the same
JPH01281098A (en) Production of optically active carboxylic acid and optically active carboxylic acid ester
JP2639651B2 (en) Process for producing optically active carboxylic acid and its enantiomer ester
JP3715662B2 (en) Process for producing optically active β-hydroxycarboxylic acid and its enantiomer ester
JP3970898B2 (en) Process for producing optically active α-methylalkanedicarboxylic acid-ω-monoester and its enantiomer diester
JPH0573396B2 (en)
JP4711367B2 (en) Method for producing optically active amino alcohol derivative
JP2928612B2 (en) Method for producing optically active amines
JPH0751533B2 (en) Process for producing optically active terphenyl derivative
JP2615768B2 (en) Optically active carboxylic acid derivative and method for producing the same
JP3741758B2 (en) Process for producing optically active glycerol derivatives
JPH0632635B2 (en) Process for producing optically active carboxylic acid and its enantiomer ester
JPH01247100A (en) Production of optically active carboxylic acid derivative
JPH0959217A (en) New optically active halogen-containing carboxylic acid derivative and its production
JP2003000294A (en) Method for producing l-phenylalanine derivative by microorganism
JPH0959211A (en) New optically active dicarboxylic acid derivative and its production