JPS61227796A - Production of optically active indoline-2-carboxylic acid - Google Patents

Abstract

PURPOSE:To obtain an optically active indoline-2-carboxylic acid or optically active indoline-2-carboxylic acid ester, by treating racemic indoline-2-carboxylic acid with a stereoselective hydrolytic enzyme, esterase or a microorganism having the above hydrolytic activity. CONSTITUTION:(R,S)-indoline-2-carboxylic acid ester of formula I (R is 1-10C alkyl, alkenyl, alicyclic hydrocarbon group, phenyl or benzyl which is substituted with hydroxyl group and/or halogen atom) is treated with a microorganism or an enzyme having stereoselective esterase activity to effect the asymmetric hydrolysis of said ester to form the optically active indoline-2-carboxylic acid of formula II. The racemic compound of formula I is resolved into the optically active compound (indoline-2-carboxylic acid of formula II) and the optically active indoline-2-carboxylic acid ester of formula III and each optically active compound can be separated from the mixture.

Description

Detailed Description of the Invention (Industrial Application Field) The present invention relates to an alkyl group or an alkenyl group having 1 to 10 prime numbers substituted with a hydroxyl group and a halogen atom, singly or simultaneously with a hydroxyl group and a halogen atom. group, unsubstituted or substituted alicyclic hypohydrogen group,
Represents an unsubstituted or substituted phenyl group or benzyl group. )
(it, s)-indoline-2-carboxylic acid ester Ie asymmetrically hydrolyzed to produce active indoline-2-carboxylic acid by the action of a microorganism or enzyme having stereoselective esterase activity. Then, an optically oily indoline-2-carbo/acid, which is a hydrolyzate, and an optically viscous indoline-2-carboxylic acid ester, which is an unreacted product, are produced from the racemic body ①, and each optically active form is separated. The present invention relates to a method for producing optically active indoline-2-carboxylic acid by optical resolution, characterized in that the sampled indoline-2-carboxylic acid is collected, and if necessary, the sampled indoline-2-carboxylic acid is further hydrolyzed to produce a counterpart body of r, and then collected.

The present invention can be achieved by selecting a stereoselective esterase to be used.
Ff
f [Can be collected.

These optically active indoline-2-carboxylic acid compounds can be used as raw materials for various pharmaceuticals. For example, ■)-indoline-2-carboxylic acid is an effective antihypertensive agent as an inhibitor of angiotensin I conversion (S)-1-1).
-Mercapto-2-oxopropylcouindoline-2
- 12 references available for carboxylic acids: Journal of Medicina, Chemistry (J, M, ed
, Chen, ).

26.894 (19819)).

(Prior Art) Regarding the production of these optically active indoline-2-carboxylic acids, a method using an optical resolution agent as shown below is known.

(Literature: JP-A-57-8140Of) [Literature: Tetrahedron Letters,
28, 1677 (however, the e) body is extracted from the mother liquor side) [Reference: Journal of Medicinal Chemistry (J, Med, Chem,), 26.1267 (Problem to be solved by the invention, @These resolving agents The optical separation method using
It has been desired to develop a method for obtaining optically active indoline-2-carboxylic acid or optically active indoline-2-carboxylic acid ester by a simple method that allows mass production of 11Cs.

(Means and effects for solving the problem) The present inventors esterified the carboxylic acid moiety of indoline-2-carboxylic acid using various alcohols, and allowed microorganisms or enzymes to act on this ester. We believe that it is possible to obtain an optically active form by performing asymmetric hydrolysis, and have been conducting repeated studies. As a result, (1) genus Bacillus, 7 strains/L/
s) or Streptomyces (Btreptomyces)
microorganisms belonging to the genus ces ), or enzymes obtained from the microorganisms, or enzymes derived from mammals (R, 8
)-indoline-2-carboxylic acid ester is asymmetrically hydrolyzed to produce ■-indoline-2-carboxylic acid ester (CB)-yne)', and then separated using an organic solvent. , to extract 9α0
-Indoline-2-carboxylic acid (9)-■ and ■)-indoline-2-carboxylic acid ester (8)-I can be collected, respectively, and the collected (S)-I can be treated by alkaline hydrolysis or enzyme treatment. (2) Pseudomonas (
pseudomonas) or Aspergillus (A
Contrary to the above (1), ω)-indoline-2-carboxylic acid (8)-4 and (6 )-yne, biline-2-carboxylic acid ester ■-■ is produced, and then separated and extracted with an organic solvent! D(8)-1 and ■-■ can be collected respectively,
The present invention was completed based on the discovery that (2)-■, which had been collected in the past, could be produced and collected by subjecting it to alkaline hydrolysis or enzymatic decomposition to produce and collect (2)-■. The present invention will be explained in more detail below.

The indoline-2-carboxylic acid esters represented by the general formula used as substrates of the present invention are generally substituted with a hydroxyl group and a halogen atom, either singly or simultaneously.
A compound which is an alkyl group or alkenyl group having 1 to 10 prime numbers, an unsubstituted or substituted alicyclic hydrocarbon group, an unsubstituted or substituted phenyl group or a benzyl group, preferably ethylene glycol, glycerol, glycerol- α−
It is an ester with toindoline-2-carboxylic acid such as monochlorohydrin, 2,8-diq4-1-brobanol, 1,8,5-pentanetriol, cyclohexanol, and benzyl alcohol.

Indoline-2-carboxylic acid ester can be obtained as follows. i.e. (It, 8)-indoline-2-
Alcohol, which serves as both a solvent and a reaction reagent, is added to carboxylic acid to obtain an indoline-2-carboxylic acid concentration of 5 to 20% (W
/V'') under strong acidity at 50°C to reflux temperature for 6 hours. Add saturated sodium hydrogen acid solution to this reaction solution, adjust the pH to 7, and add acetic acid. Ethyl, chloroform.

Extracting with a hydrophobic organic solvent such as methylene chloride, hexane, etc. and further concentrating yields high purity (IL).

8) -Indoline-2-carboxylic acid ester I 2>
(can get.

Examples of microorganisms that have a stereoselective esterase that asymmetrically hydrolyzes racemic body (I) to produce ToI include the genus Bacillus.

Aspergillus JII
4 or Streptomyces (3treptom) rce
There are microorganisms belonging to the genus Bacillus subtilis.
) I FO8018, Aspergillus meleus (A
spergillus melleus) I FO4
420e Streptomyces griseus (Strep
tomyces griseus) IFO8858 Galuru.

For culturing these microorganisms, any nutrient medium in which the microorganisms can grow can be used, for example, a nutrient medium containing glucose, peptone, yeast extract, meat extract, etc. is used. The culture temperature in the medium is 10 to 40°C, preferably 25 to 85°C, the pH is 8 to 8, preferably 6 to 7, and the culture is aerobic, usually for 24 to 48 hours. .

In the asymmetric hydrolysis reaction of indoline-2-carboxylic acid ester by a chiral organism, there is a method in which the compound 1 is added to the medium at the same time as the start of the culture, and hydrolysis is carried out in parallel with the culture. Add the compound (2) to the obtained culture solution containing bacterial cells, or after culturing, centrifuge 1lIl or filtrate and collect the resulting bacterial cells! There is a method of hydrolyzing the bacterial cells by contacting them with a compound in a suspension of bacterial cells made into a fresh solution, but it is preferable to concentrate the bacterial cells by centrifugation or filtration, and then remove the cells to a high concentration. The method of forming a suspension and adding compound (1) to this suspension is superior from the standpoint of yielding the product (6) after the reaction.

The solubility of the compound (2) in water is generally low, but if it is stirred, it will not cause any problem as it will remove the unreacted button. Further, organic solvents such as acetonate and methanol, surfactants, etc. may be added to the extent that they do not interfere with the reaction.

Reaction conditions are fM degrees 10-40°C, preferably 25-85
℃ range, and the pH is 5 to 8, preferably 6.5 to 7.
．． The reaction time varies depending on the ratio of the substrate and the amount of microbial cells, but it is sufficient to stop when the molar ratio of unreacted ester to product carboxylic acid reaches 50%. however,
From the viewpoint of bacterial reaction activity, it usually reaches 50% in 24 to 72 hours.
It is desirable to determine the amount of substrate added so that

As a method using an enzyme, a crude enzyme obtained by crushing the microbial cells, followed by ammonium sulfate fractionation or acetone treatment, or a purified enzyme obtained by performing a column chromatography operation can be used. Commercially available enzymes include biobrase AL-15 (origin: Bacillus subtilis, manufactured by Naga@Sangyo), grotease "
Aikuno' P (origin: Aspergillus meleus, Ohno Pharmaceutical-all), actenase E (origin: Streptomyces griseus, manufactured by Kaken! $1! Yakuhin 6m), Steapsin (pig pancreas, manufactured by Wako Tsumugi Pharmaceutical ■) , lipase L8126
(pig pancreas, manufactured by Sigma), pancreatic digestive enzyme TA (manufactured by Amano Rakutoki), etc. can be used. Furthermore, when producing (g)-M, for example, riboprotein lipase (L, P, L
, Amano8. Origin: Pseudomonas aeruginosa, Ohno Pharmaceutical ■IM), lipase AP-6 (origin: Aspergillus niger, Ohno Pharmaceutical ■), etc. can also be used. In the asymmetric hydrolysis reaction, two semiforms of the substrate are suspended in a reaction solution at a concentration of 2 to 80% (W/V), and # element is passed through, for example, as a weight ratio of enzyme and substrate of 1: 5 or t
: ioo.

and at a temperature of 10-40°C, preferably 25-40°C.
The reaction was carried out in the range of 85°C, and the amount of carbo/acid produced and the decrease in carboxylic acid ester were measured by high-performance liquid cocomatography, and when the molar ratio of 11 and 9 in the reaction solution reached 50%. It's okay if you stop reacting. In addition, it does not matter if the pIII range during hydrolysis is 4 to 8.5, but as the hydrolysis reaction progresses, the pH in the reaction solution tends to be acidic. 6～
It is desirable to keep it at 7. Furthermore, the above asymmetric hydrolysis reaction can be repeated by immobilizing microbial cells or B8, for example.

To remove microorganisms, after asymmetric hydrolysis using enzymes, the method of separating ■ and ■ in the reaction solution is to adjust the pH of the reaction solution.
After adjusting to 7, I\1'' acid ether, chloroform,
By extracting only the optically active indoline-2-carboxylic acid ester (I) with a hydrophobic organic solvent such as methylene chloride or hexane, it can be easily separated from the hydrophilic optically active indoline-2-carboxylic acid I.

The optically active indoline-2-carboxylic acid ester obtained by separation can be obtained as an ester with high optical purity by concentrating as it is, but it can be further purified with K as follows to obtain optically active indoline/-2-carboxylic acid. be able to. That is, optically active indoline-2-carboxylic acid ester ■)-1 or ■
If -■ is subjected to alkaline hydrolysis at room temperature for 10 minutes to 2 hours in the range of 1) H9S11, then -mi or ■-mi will be produced, respectively.

In addition, for enzymes that have the ability to hydrolyze (8)-, L or ■-chemistry, for example ■)-1, ribobutin lipase Amano 8 was used, while for (2)-1, stearpsin was used. If hydrolysis is carried out under the conditions of hydrolysis using the above-mentioned # element, ①)-Wataru or ①-■ can be obtained, respectively.

The pH of the hydrolyzed solution obtained in this way is adjusted to 4 to 6, preferably around 6.0, and then extracted with a solvent such as methylene chloride and ethyl acetate. By crystallizing it in a medium, 0)- or (2)-methods with high optical purity can be obtained.

On the other hand, as mentioned above, the optically active indoline-2-carboxylic acid remaining in the aqueous layer during extraction separation also has 1) H4-6
, preferably adjusted to around 5.0, and then subjected to similar extraction and purification operations to obtain (2)- or (B)- of high optical purity.
You can easily get the view.

In the asymmetric hydrolysis reaction using microbial cells, the cells remain in the aqueous layer after extraction and separation with an organic solvent, but if the pH is subsequently lowered and an organic solvent extraction operation is performed, the target optically active indoline- There is no problem in collecting 2-carboxylic acid. In addition, after removing microbial cells by centrifugation or filtration, indoline-2
-Carboxylic acid and indoline-2-carboxylic acid ester can be separated and extracted.

(Examples) Hereinafter, the present invention will be specifically explained with reference to Examples, but the present invention is not limited to these Examples.

Example 1 Production of (R,8)-indoline-2-carboxylic acid glycerol ester Ia. The condensation reaction was carried out at a temperature range of 8 hours.

After the reaction, the reaction mixture was once cooled, and then sodium bicarbonate and saturated aqueous sodium bicarbonate were added to adjust 1) H to 7.0-. Next, extraction was performed 8 times (total 1.51 times) with 500 ml of ethyl acetate.
The ethyl acetate layer was washed with 100 Mt of water, dehydrated with anhydrous sodium sulfate, and further concentrated under reduced pressure to obtain (R,5)-1.
A was obtained with a yield of 57.7f and 79%.

In a similar manner, the following substrate (R,8)-indoline-2-carboxylic acid ethylene glycol ester was prepared by adding 5 times the amount of alcohol to 50 fK of (R,S)-indoline-2-carboxylic acid. (Ib) 50.2 f Yield 79.0% (R,8)-Indoline-2-carboxylic acid glycerol α-monochlorohydrogen. Dorin Esthe/L/(IC)45.
49 Yield 68.0% (R,8)-Indoline-2-carboxylic acid 2,8-cyclogropanol ester (Id) 28.19 Yield
88.4% (R,8)-indoline-2-carboxylic acid 1°8.5-
Pentanetriol ester (Ie) 59.5f Yield 78.8% (R,S)-indoline-2-carboxylic acid 1,4-cyclohexanediol ester (I f) 56.4f
Yield 70.8% (R,8)-Indoline-2-carboxylic acid benzyl alcohol ester (Ig) 49.7 F Yield 64
．． 0% was prepared. In addition, Ib, Ic, Id, If, Ig
During the purification, methylene chloride was used instead of ethyl acetate as the extraction solvent.

Example 2 Substrate (R
,S)-indoline-2-carboxylic acid and stearpsin 0.21 were added, and the pH was adjusted to 7 with 2N NaOH solution.
．． The asymmetric hydrolysis reaction was carried out at 88° C. for 6 hours while stirring. This reaction solution was mixed with 200% of ethyl acetate.
The extraction operation was performed twice (600,111 tons in total), and the ethyl acetate layer was dehydrated with anhydrous sodium sulfate and concentrated under reduced pressure to obtain a specific optical rotation [α]. Viscous syrup with +16.2° (C=1.Oe ethanol) ■) - 4.6flR, 8) -
A yield of 92% was obtained.

IHNMR (90MHz) The measured values were as follows.

IHNMR (Mean d4) δppm: 8.2
~8.45 (2H), 4.7 to 8.5 (9
H), 6.5 A-7,1 (4H1mtAr-) 0.1M dipotassium hydrogen phosphate solution 80- was added to the obtained (8)-Ia 4.111, and 2NN was added at 88°C.
The pH was maintained at lO by dropping aOH and hydrolysis was carried out for 1 hour. After that, the reaction solution was diluted with IN hydrochloric acid to 1)H
Adjust to
m) An extraction operation was performed. After further dehydration with anhydrous sodium sulfate, it was concentrated under reduced pressure, and the dried product was dissolved in acetone-hexane (51R
When it recombines at t-1*), the specific optical rotation [α), +82.
5' (C=1.0. dimethylformamide) (
Literature value, J, Med, Chem, ).

■, 894 (198B), (α], +84.5° ((
=l,Q,dimethylformamide))<8)-Indoline-2-carboxylic acid (S)-2
．． 61g1 18)! , a yield of 76% was obtained.

The in NMR (90M) measurement values were as follows.

IHNMR (pH80-d6) δppm: 2.
85-8.45 (2H), 4.10-4.85
(IH).

6.40-7.05 (4H, m, ArV/l).

On the other hand, the aqueous layer after the first ethyl acetate extraction was adjusted to pH with IN hydrochloric acid.
5.0, repeat the extraction 8 times (total 600 mj) using 200 rnt of ethyl acetate, and perform the same operation as in the case of (■)-mi below, and (8)-mi is 8.0211
(Yield 88% from (R-,8)-Ia) was obtained.

The value of specific optical rotation was [α)D-29.2° (C=1.0.dimethylformamide).

Examples 8 to 9 The same operations as in Example 2 were carried out by changing the enzyme, and the results shown in Table 1 were obtained.

In the table, Examples 8, 4, 5, 6.7 have only 8 bodies, 8.9
is an example in which only 8 bodies were asymmetrically hydrolyzed.

Enzyme reaction conditions Two substrates (几, 5) -11101゜Enzyme each 0
．． 5N, 0.1M phosphate buffer, pH 7.0 100m each
, 88°C, stirring with stirrer, reaction for 6 hours, pH controlled at 7.0.

DMF Nidimethylformamide (blank below) Examples 10 to 15 Steapsin was used as the enzyme, and ethylene glycol ester of (R,8)-indoline-2-carboxylic acid I was used as the substrate.
b, glycerol α-monochlorohydrin ester Ic
e 2.8-cyclo-1-7” lovanol ester Id, 1.8.5-pentanetriol ester I?
, 1,4-cyclohexanediol ester If
The same operation as in Example 2 was performed except that Hensyl alcohol ester Ig was used, and the results shown in Table 2 were obtained.

Enzyme reaction conditions: 1 each of two substrates (1, stearpsin 0.5Ii
, 0.1M phosphate buffer, pH 7.0 each 100-088
℃, stirred with a stirrer, reacted for 6 hours, controlled at p■7, 0.

(The following is a blank space) Example 16 A nutrient liquid medium having the following composition was prepared, dispensed into two Sakaguchi flasks in 400 m portions, and sterilized at 120° C. for 15 minutes.

[Medium composition]

Glucose 4%, yeast extract 0.8%, meat extract 0
．． 8%, peptone 0.8%, ammonium phosphate 0.
2%, potassium phosphate 0.1% (1) H6, 8) Separately, #t of Pseudomonas aeruginosa IFO8080 was precultured in a medium with the same composition. Bacterial liquid 10-
was inoculated into the culture medium and shaken at 80°C for 124 hours. A total of 5 cells were cultured, and a culture solution needle 21 was obtained. This culture solution was centrifuged and the bacterial cells were collected. This bacterial body is 0.1M
Phosphate buffer (pH 7-0) 200m/WN! cloudy,
! 2.0 g of glycerol (R, 8) -I:/thrine-2-carboxylic acid Ia was added. This is 500+
d Adjust the pH to 7 with IN NaII solution under stirring in a valley vessel.
．． The reaction was carried out at 80°C for 12 hours while adjusting the temperature to 0. After the reaction, the supernatant obtained by centrifugation was extracted and separated four times (total of 8001 R1) with 200 ml of ethyl acetate, and then the same operation as in Example 2 was performed to obtain the results shown in Table 8. Ta.

Example 17^21 Pseudomonas aeruginosa of Example 17 IF0 1
8180. Bacillus subtilis in Example 19 was cultured in the same manner as in Example 16, and the microorganisms of the genus Aspergillus in Examples 18 and 20 were cultured using 8.0% glucose, 1.0% polypeptone, and eth)ff-"?s. 0.5%, ammonium phosphate 0.2%, potassium phosphate 0.1%
The same procedure as in Example 16 was carried out except that the culture medium (I)H6,5) was used and the temperature was 28°C.

After culturing each strain, the bacterial cells were collected by centrifugation for Pseudomonas aeruginosa and Bacillus subtilis, and by filtration for Aspergillus microorganisms, and suspended in 0.1M phosphate buffer pH 7, 0,200 dK. Example 1
Asymmetric hydrolysis reaction using microorganisms, extraction, and purification were performed according to 6, and the results shown in Table 8 were obtained.

(Effect of the invention) According to the present invention, it is possible to appropriately select and use a hydrolytic enzyme esterase having stereoselectivity or a microorganism having the same hydrolyzing ability, (R,,8)-indoly/-2 −
Carboxylic acid ester Optically active form of the ester (6
) form or ■) form, or optically active indoline-2-carboxylic acid, (6) form or (8) form, respectively, can be obtained arbitrarily.

Patent applicant Kanebuchi Chemical Industry Co., Ltd. Agent Patent Attorney Makoto Asano - Procedural Amendment Showa tσ year! 21st of the month

Claims (16)

[Claims] (1) General formula ￥1￥ (R, S) - ▲ There are mathematical formulas, chemical formulas, tables, etc. ▼￥ I
￥ (wherein B is an alkyl group or alkenyl group having 1 to 10 carbon atoms substituted singly or simultaneously with a hydroxyl group and a halogen atom, an unsubstituted or substituted alicyclic hydrocarbon group,
Represents an unsubstituted or substituted phenyl group or benzyl group. )
Asymmetrically hydrolyzing (R,S)-indoline-2-carboxylic acid ester ￥ I ￥ represented by the structural formula ￥ II ￥
＾＊ ▲There are mathematical formulas, chemical formulas, tables, etc. ▼￥II￥＾＊ By acting on a microorganism or an enzyme that has stereoselective esterase activity to produce optically active indoline-2-carboxylic acid, racemic ¥ I ¥
The optically active compound indoline-2-carboxylic acid (￥I
I￥＾*) and optically active indoline-2-carboxylic acid ester represented by the general formula ￥ I ￥＾* ▲Mathematical formulas, chemical formulas, tables, etc.▼￥ I ￥＾* (R is the same as above) Optically active indoline produced by optical resolution, characterized by optical resolution and separation and collection of each optically active substance.
Method for producing 2-carboxylic acid. (2) Optically active indoline-2-carboxylic acid is optically active (R)-indoline expressed by the structural formula (R)-\II\ ▲There are mathematical formulas, chemical formulas, tables, etc.▼(R)-\II\ 2-carboxylic acid, optically active indoline-2-carboxylic acid ester has the general formula (S) - ￥ I ￥ (S) - ▲ Numerical formulas, chemical formulas, tables, etc. ▼ (S) - ￥ I ￥ (R The manufacturing method according to claim 1, which is (S)-indoline-2-carboxylic acid ester represented by (same as above). (3) The microorganism or enzyme is Bacillus (Bacillus).
s) genus Aspergillus or Streptomyces
The manufacturing method according to claim 1 or 2, which is a microorganism belonging to the genus or an enzyme derived from the microorganism. (4) The manufacturing method according to claim 1 or 2, wherein the enzyme is an enzyme derived from a mammalian organ. (5) Optically active indoline-2-carboxylic acid ￥IIＥ＾*
is the structural formula (S)-\II\ (S)-▲There are mathematical formulas, chemical formulas, tables, etc.▼(S)-\
It is optically active (S)-indoline-2-carboxylic acid represented by
There are mathematical formulas, chemical formulas, tables, etc. ▼ The optically active (R)-indoline-2-carboxylic acid ester represented by (R)-\I\ (R is the same as above) as described in claim 1. Production method. (6) The microorganism or enzyme is of the genus Pseudomonas or Aspergillus (A
6. The production method according to claim 1 or 5, which is a microorganism belonging to the genus Spergillus or an enzyme derived from the microorganism. (7) General formula ￥ I ￥ R is -CH_2-C(OH)C
The manufacturing method according to any one of claims 1 to 6, wherein H_2OH is used. (8) The manufacturing method according to any one of claims 1 to 6, wherein R in the general formula ¥ I ¥ is -CH_2CH_2OH. (9) General formula ¥ I ¥ (R, S) - ▲ Numerical formulas, chemical formulas, tables, etc. are available ▼ ¥ I
¥ (wherein, R is an alkyl group or alkenyl group having 1 to 10 carbon atoms substituted singly or simultaneously with a hydroxyl group and a halogen atom, an unsubstituted or substituted alicyclic hydrocarbon group, an unsubstituted or substituted phenyl group or a benzyl group) (R,S)-indoline-2-
Asymmetrically hydrolyzing carboxylic acid ester ￥ I ￥,
Structural formula ￥II￥＾＊ ▲Mathematical formulas, chemical formulas, tables, etc. are available▼￥II￥＾＊ A microorganism or an enzyme having stereoselective esterase activity that produces optically active indoline-2-carboxylic acid is acted on. Therefore, racemic compound ￥ I ￥
The optically active compound indoline-2-carboxylic acid (￥I
I￥＾*) and the optically active indoline-2-carboxylic acid ester represented by the general formula ￥ I ￥＾* ▲Mathematical formulas, chemical formulas, tables, etc.▼￥ I ￥＾* (R is the same as above) Split, separate and collect each optically active substance,
Optically active indoline-2-carboxylic acid obtained by optical resolution is characterized by further hydrolyzing ¥I ¥^* to produce and collect optically active indoline-2-carboxylic acid, which is the enantiomer of compound II^*. Acid production method. (10) Optically active indoline-2-carboxylic acid has the structural formula (R)-\II\ (R)-▲numeric formula, chemical formula, table, etc.▼(R)-\
II￥ is an optically active (R)-indoline-2-carboxylic acid represented by The manufacturing method according to claim 9, which is a (S)-indoline-2-carboxylic acid ester represented by ▼(S)-\I\ (R is the same as above). (11) The microorganism or enzyme is Bacillus (Bacillus).
us) genus, Aspergillus
Genus or Streptomyces
11. The production method according to claim 9 or 10, wherein the method is a microorganism belonging to the genus A. ) or an enzyme derived from the microorganism. (12) The production method according to claim 9 or 10, wherein the enzyme is an enzyme derived from a mammalian organ. (13) Optically active indoline-2-carboxylic acid ￥II＾
* is the structural formula (S)-\II\ (S)-▲There are mathematical formulas, chemical formulas, tables, etc.▼(S)-\
It is optically active (S)-indoline-2-carboxylic acid represented by
There are mathematical formulas, chemical formulas, tables, etc. ▼(R)-\ I\ (R is the same as above) Optically active (R)-indoline-2-carboxylic acid ester according to claim 9 Production method. (14) The microorganism or enzyme is Pseudomonas (Pseudomonas)
domonas) or Aspergillus (Aspergillus)
14. The production method according to claim 9 or 13, which is a microorganism belonging to the genus P. illus or an enzyme derived from the microorganism. (15) General formula ￥ I ￥ R is -CH_2-C(OH)
The manufacturing method according to any one of claims 9 to 14, wherein CH_2OH is used. (16) General formula ￥ I ￥ R is -CH_2CH_2OH
A manufacturing method according to any one of claims 9 to 14.