JPS58126792A - Preparation of optically active substance by microorganism - Google Patents

Abstract

PURPOSE:To prepare an optically active alcohol and/or a carboxylic acid useful as a herbicide, insecticide, etc., by reacting a microorganism or a treated substance thereof with a racemate of a carboxylic acid having an asymmetric carbon atom. CONSTITUTION:A microorganism belonging to the genus Glomerella or Schizophyllum is cultivated in the presence of a racemate of a carboxylic acid expressed by formulaI(X is a group having a ring structure which may be substituted; R<1> is H or alkyl; R<2> is -O-X, -(CH2)nCOOR<1> and a group other than hydrogen; n is an integer 0-10) or the microorganism or a treated substance thereof is reacted with the compound expressed by formulaIto give an alcohol, having the optical activity, and expressed by formula II (X, R<2> and n are same as described above) or a carboxylic acid, having the optical activity, and expressed by formulaIor an ester thereof.

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention involves culturing a racemic compound of a carboxylic acid compound or a carboxylic acid ester compound having an asymmetric carbon with a microorganism, or allowing a culture of the microorganism or a processed product thereof to act on the alcohol compound having scientific activity. and/or a method for producing a carboxylic acid compound or carboxylic acid ester compound having scientific activity.

Previously, one of the inventors, Nakajima et al., converted carboxylic acid compounds without asymmetric carbons, such as 2,4-dichlorophenoxyacetic acid, into 2(2,4-dichlorophenoxy)ethanol by a microbial culture. I found out. [Chemical Pharmaceutical Bulletin 21(3)
See pages 671-673 (1973)] Based on these findings, the present inventors have conducted extensive research, and as a result, using racemic compounds of carboxylic acid compounds or carboxylic acid ester compounds having asymmetric carbon atoms as starting materials, various It is further scientifically proven that a carboxylic acid compound or carboxylic acid ester compound that has no scientific activity is converted into an optically active alcohol compound by culturing with a microorganism or acting on a culture of the microorganism or a processed product thereof. It was discovered that a carboxylic acid compound or carboxylic ester compound having activity can be obtained, and the present invention was achieved.

The starting material used in the present invention has the formula (wherein, X is a group having an optionally substituted cyclic structure, R1 is hydrogen or an alkyne group, R2 is -O-X in the formula,
-(CH2)n-COOR1 and groups other than hydrogen; n represents an integer of 0 to 10; ) is a racemic compound of a carboxylic acid compound or carboxylic acid ester compound represented by , but is not particularly limited to these groups.

R1 is hydrogen or an alkyl group, and R2 bonded to an asymmetric carbon is -O-X, -(CH2)n-CO in the formula
Any group other than OR1 and hydrogen may be used, such as an alkyl group, an alkoxy group, an amino group, a hydroxyl group, a phenyl group, chlorine, fluorine, bromine, iodine, a nitro group, a thiol group, an aldehyde group, etc. It is not particularly limited to these groups.

Examples of microorganisms used in the reaction include Glomerella (
Gloeosporium olivarum arum belongs to the genus Glomerella.
liuarumAlm) (obtained from the Plant Pathology Laboratory, Faculty of Agriculture, Kagawa University), Gloeosporium persimmon (Gloeo
sporium kaki) (obtained from the Fermentation Research Institute, IFO 6445), Glomerella cingulata, and Schizophyllum commune belonging to the genus Schizophyllum.
um comune) (obtained from the Mycology Laboratory, Forestry Research Institute, Ministry of Agriculture, Forestry and Fisheries).

To carry out the present invention, microorganisms belonging to the genus Glomerella or Schizophyllum are left standing or shaken in an appropriate medium containing any starting material, and after culturing and reacting for a certain period of time, the culture solution is Separate into bacterial cells and furnace liquid. This reaction may be carried out by adding a starting material to a culture solution of microorganisms or a processed product thereof, and allowing the mixture to stand or be shaken.

Next, the target substance can be obtained by performing extraction and purification operations on the furnace liquid. For example, the pH of the furnace solution is adjusted to alkaline, and then extracted with an organic solvent such as ether. After washing the extract, an alcohol compound or a carboxylic acid ester compound having the desired optical activity can be obtained by performing operations such as vacuum concentration and chromatography.

On the other hand, after the pH of the reactor solution residue after extraction with an organic solvent is adjusted to the desired value, it is extracted with an organic solvent such as ether. After washing the extract, a carboxylic acid compound having the desired optical activity can be obtained by vacuum concentration, chromatography, etc.

The alcohol compound having the desired optical activity has the formula (wherein,
X, R2, and n have the same meanings as above. ),
Further, the optically active carboxylic acid compound or carboxylic acid ester compound has the formula (wherein, X, R1, R2, and n have the same meanings as above).

).

Depending on the physical properties of the starting material and target material, extraction, separation,
It is preferable to change the conditions for purification etc. as appropriate.

The optically active substance obtained in the present invention is useful as a herbicide, insecticide, etc.

Example 1 90 g of peptone, 3.6 g of magnesium sulfate heptahydrate,
As a publication substance, 2-
After adding 4.25 g of phenoxypropionic acid, the pH
5.5, inoculated with Gleosporium olivarum arum, and then dispensed into a 2-liter jar fermenter, followed by aeration-stirring culture at 27-28°C for 5 days.

The ventilation amount is 1.2 l/min and the propeller rotation speed is 450 rpm.

After the culture is completed, the culture reaction solution is filtered by suction to separate the bacterial cells from the filtrate.

Next, the pH of the furnace solution was adjusted to 9.2 with 10% potassium hydroxide, and then extracted with 3 liters of ether. This ether extract was washed repeatedly with saturated brine until the washings became neutral, and then dried with nitric oxide. Next, the ether extract was concentrated under reduced pressure, and this concentrated liquid was developed using chloroform in silica gel column chromatography, and the eluate was dried under reduced pressure to obtain 0.8 g of 2-phenoxypropanol having high activity.

Specific optical rotation [α] 25+13.7° (C0.00876 methanol) The infrared absorption spectrum, nuclear magnetic resonance spectrum, and mass spectrum of this substance were consistent with the racemic form of 2-phenoxy-propanol prepared separately.

On the other hand, the ether-extracted residue was adjusted to pH 1.5 with hydrochloric acid, then further extracted with 3 liters of ether, the extract was washed with saturated saline, and then dried over (2) nitric chloride.

The mixture was then concentrated under reduced pressure, and this concentrated solution was developed using chloroform in silica gel column chromatography, and the eluate was dried under reduced pressure to obtain 1 g of optically active 2-phenoxypropionic acid.

Specific optical rotation: [α]25+16.9° (C0.0025 acetone) The infrared absorption spectrum, nuclear magnetic resonance spectrum, and mass spectrum of this substance are consistent with the starting material 2-phenoxy-propionic acid (racemic form). did.

Example 2 After adding 3 g of 2-phenoxy abbreviation as a starting material to 9 liters of peptone medium having the same composition as in Example 1, the pH was adjusted to 5.5, and 300 ml was dispensed into 1000 ml flasks. - Inoculate C. singulata and culture with shaking at 27-28°C for 7 days.

This culture reaction liquid is suction filtered and separated into bacterial cells and fermentation liquid.

Next, the pH of the furnace solution was adjusted to 9.5 with 10% sodium hydroxide, and then extracted with 3 liters of ether. This extract was washed repeatedly with saturated saline until the washing became neutral, and then dried with nitric chloride. Next, it was concentrated under reduced pressure, and this concentrated solution was developed using chloroform in silica gel column chromatography, and the eluate was dried under reduced pressure to obtain 0.9 g of optically active 2-phenoxy-n-butyl alcohol.

Specific optical rotation: [α] 25-4.6 (C0.00245 methanol) The infrared absorption spectrum of this substance matched that of a racemic form of 2-phenoxy-n-butyl alcohol prepared separately.

On the other hand, after adjusting the ether-extracted residue to pH 1.2 with hydrochloric acid, further extraction was performed with 3 liters of ether, and this extract was washed with saturated saline, and then dried with nitric acid. Next, it was concentrated under reduced pressure, and this concentrated solution was developed using chloroform using silica gel column chromatography.
The eluate was collected and dried under reduced pressure to obtain 1.34 g of optically active 2-phenoxybutyric acid.

Specific optical rotation: [α] 25 + 10.1° (C0.00251
4acetone) The infrared absorption spectrum, nuclear magnetic resonance spectrum, and mass spectrum of this substance are based on the starting material 2-acetone.
It was consistent with phenoxybutyric acid (racemic form).

Example 3 In the same manner as in Example 1.2, various starting materials were added to a peptone medium and reactions were attempted under the following conditions to obtain the target substance.

Claims (1)

[Claims] 1) Formula (wherein, X is a group having an optionally substituted cyclic structure, R1 is hydrogen or an alkyl group, R2 is -O-X in the formula,
-(CH2)n-COOR1 and groups other than hydrogen; n represents an integer of 0 to 10; ) is cultured with a microorganism or treated with a culture of the microorganism or its treated product, and the racemic form of the carboxylic acid compound or carboxylic acid ester compound represented by the formula (wherein X, R2, n are A method for producing an alcohol compound represented by the formula (having the same meaning as above) and/or a carboxylic acid compound represented by the formula having scientific activity (wherein X, R1, R2, n have the same meaning as above); Method for producing carboxylic acid ester compounds. 2) The method according to claim 1, wherein the microorganism belongs to the genus Glomerella. 3) The microorganism is Schizophyllum.
) The manufacturing method according to claim 1, wherein the microorganism belongs to the genus A.