JPS59130188A - Biochemical preparation of optically active alpha-cyano-3-phenoxybenzyl alcohol - Google Patents

Abstract

PURPOSE:To prepare the titled compound useful as a component of an insecticide, pyrethroid, by the asymmetric hydrolysis with an esterase produced by a specific microorganism. CONSTITUTION:The esterase produced by a microbial strain capable of conducting preferential asymmetric hydrolysis of the carbonate of (S)-alcohol when made to contact with the carbonate ester of (R,S)-alpha-cyano-3-phenoxybenzyl alcohol of formula (R is 1-8C alkyl, alkenyl or alkynyl), e.g. Arthorobacter simplex IFO 3530, is made to contact with the carbonate ester of said (R,S)-alcohol at <=7 pH, preferably 3.5-6.5 pH to resolve the above racemic mixture into the carbonate ester of an optically active alpha-cyano-3-phenoxybenzyl alcohol rich in (S)-isomer and the carbonate ester of its antipode.

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to a biochemical method for producing optically active α-cyano-8-phenoxybenzyl alcohol. More specifically, the general formula (wherein k is an alkyl group having 1 to 8 carbon atoms,
It represents an alkenyl group having 1 to 8 carbon atoms or an alkynyl group having 1 to 8 carbon atoms. Production of microorganisms having the ability to preferentially asymmetrically hydrolyze the carbonate ester of monolithic alcohol (S) by acting on the carbonate ester of (R,S)-α-cyano-8-phenoxybenzyl alcohol represented by The esterase is allowed to act on the carbonate ester of the (,R,s)-alcohol at pH 7 or lower, and this is asymmetrically hydrolyzed to form (S)-alcohol into optically active α-cyano-3-phenoxybenzyl alcohol. , 4 relates to a biochemical method for producing optically active α-cyano-3-phenoxybenzyl alcohol by obtaining its enantiomer carbonate ester.

α-Cyano-3-phenoxybenzyl alcohol is known as a novel alcohol component of a group of ester compounds called synthetic pyrethroids, which have excellent insecticidal activity.

The α-cyano-3-phenoxybenzyl alcohol is
Since it has an asymmetric carbon at the α-position, two types of optical isomers exist, and the insecticidal efficacy as an ester is (S).
Alcohol is far superior to the body in terms of its response (Kosuke Yoshioka, Organic Synthetic Chemistry, Vol. 38, No. 12, 1)
pp. 151-1162 (1980). Therefore, it is possible to optically resolve (R,S')-α-cyano-3-phenoxybenzyl alcohol obtained by ordinary synthetic chemical production methods in an industrially advantageous manner to obtain (S) monomer. Development is desired.

However, since α-cyano-8-phenoxybenzyl alcohol is an unstable compound, its optical resolution is not easy, and currently known optical resolution methods for this alcohol require complicated steps and expensive optically active reagents. The current situation is that this is necessary.

The present inventors have found that (S)-α can be an industrially advantageous production method.
- As a result of repeated research to find a method for producing cyano-3-phenoxybenzyl alcohol, racemic Kyubechi (R,
By biochemically asymmetrically hydrolyzing the carbonate ester of S)-α-cyano-8-phenoxybenzyl alcohol as a raw material, optically active α-rich in (S)
It was discovered that cyano-3-phenoxybenzyl alcohol and its enantiomer carbonate ester can be efficiently separated, and after further various studies, the present invention was completed.

That is, the present invention acts on the carbonate ester of (R,S)-α-cyano-8-phenoxybenzyl alcohol to produce (S)
Among the esterases (as used in the present invention) produced by microorganisms that have the ability to preferentially asymmetrically hydrolyze carbonic esters of monoalcohols, the term esterase refers to esterases in a broad sense including lipases. ) is allowed to act on the carbonate ester of the racemic alcohol at pH 7 or below, and this is asymmetrically hydrolyzed to form (S) an optically active α-cyano-8-rich monomer.
Optically active α-cyano-3 produced by splitting phenoxybenzyl alcohol and its enantiomer carbonate ester
- Provides a method for producing phenoxybenzyl alcohol.

Next, the method of the present invention will be described in detail. In the present invention, (R,S)-α-cyano-3- used as a raw material
Phenoxybenzyl alcohol can be easily produced by a conventional method, for example, by reacting (R,S)-α-cyano-3-phenoxybenzyl alcohol with an alkyl chlorocarbonate. Examples of the alkyl chlorocarbonate include commercially available methyl chlorocarbonate,
Examples of microorganisms that produce the esterase used in the present invention include chloroethyl carbonate, chloropropyl carbonate, and the like.
) Any microorganism can be used as long as it produces an esterase capable of preferentially asymmetrically hydrolyzing the carbonate ester of monolithic alcohol. Examples of such microorganisms include Arthrobacter
r) genus, Achromobacter
er) genus, Pseudomonas
Genus, Chromobacterium
um) genus, Bacillus genus,
Brevibacterium
Genus, Genus Nocardia, Genus Rhodotorula, Genus Candida, )') : I Te' 11
/ v (Trichoderma) genus, Rhizopus (
Rhizopus, Mucor, Aspergillus, Geotricnm, Aureobasidium, Hansenula, T'or
Examples include microorganisms belonging to the genus P. ulopsis.

Specific examples of this microorganism include the following bacterial cells.

(1) Arthrobacter simpletsucus IFO
-8580 Arthrobacter simplex (2
) ArCCney 21
910Achromobacter Sp・(8)
Pseudomonas fluorescens IFO'-
8081Pseudαnonas fluoresc
ens(4) Chromobacterium viscozam
AATCC-6918Cranobacterium
viscosum (5) Bacillus licheniformis IFO-12197Bacillus
licheniformis (6) Brevibacterium ammoniagenes IFO-12072Bre
vibacterium ammoniagene
s(7) Tukartia erythropolis I
FO-1282ONocardia erythr
opol 1s(8) Rhoclotorula m1n
uta var, texensisIFO-0
879 (9) Candida utilis IFO-
1086Candida utifices (10
) l-Lycoderma viride IFO-4
847 Trichoderma viride (11
) Rhizopus chinensis IFO-47
87 Rhizopus chinensis (12
) Mucor Javanix IFO-4572
Mucor javanicus (13) Aspergillus var asper I FO
-5824 Aspergillus var,
asper (14) Geotrichum candium
IFO-5868 Geotricum can
didum (15) Aureobasidius pullulans I FO-4464Aureobas idi
um pul Jul ang (16) Hansenula anomala IFO-0707Han
senula anomala (17) Torulopsis chiandida 1po-0880Torul
opt is candida. Both of these strains are produced by the Institute of Fermentation (IFO) or A.
merican Type CultureColle
ction (ATC:C) and can be obtained from this institution.

Furthermore, some esterases derived from microorganisms are commercially available, and these commercially available enzymes can also be used for the purpose of the present invention.

Commercially available enzymes that can be used in the present invention include those shown below.

When carrying out the method of the present invention, the formula (
The asymmetric hydrolysis of the carbonate ester of R,S)-α-cyano-3-phenoxybenzyl alcohol can be carried out using a culture solution in which the above-mentioned microorganisms have been cultured, a culture solution, an aqueous suspension of bacterial cells, or a treated product thereof, such as crude esterase. This is carried out by mixing an aqueous solution containing purified esterase and the carbonate ester of (R,S) monolithic alecole, and stirring or shaking the mixture. If necessary, a non-ester surfactant may be added. It is also possible to immobilize the enzyme and use it.

At this time, the reaction temperature is suitably 10 to 65°C, particularly preferably 20 to 50°C. The reaction time is usually 8 to 48 hours, but the reaction temperature may be increased or
The reaction time can also be shortened by using a highly active enzyme.

It is important that the reaction pH is 7 or less, and preferably maintained within the range of pH 8.5 to pH 6.5. The reaction product in the hydrolysis reaction of the carbonate ester of (R,S)-α-cyano-8-phenoxybenzyl alcohol represented by the above general formula is α-cyano-3-phenoxybenzyl alcohol, represented by the general formula ROH. Since these are alcohol and carbon dioxide, the pH hardly changes during the reaction due to these reaction products, and it is extremely easy to maintain the pH during the reaction.

The concentration of the carbonate ester substrate used is based on the reaction solution.
Range of 1 to 8 Q w/w%, preferably 5 to 3
5 w/w%.

After carrying out the asymmetric hydrolysis reaction in this manner, phenoxybenzyl alcohol and unreacted carbonate ester of the enantiomer alcohol are separated and recovered.

For example, the reaction completed solution is extracted with an organic solvent such as ether, benzene, or toluene, and this extract is subjected to column chromatography using silica gel or the like and eluted with a cyclohexane-ether (95:5) solution. The optically active α-cyano-8-phenoxybenzyl alcohol and the unreacted enantiomer carbonate can be separated and obtained.

The unreacted carbonate ester thus separated and recovered can be turned into shrimp by contacting it with a basic compound such as ammonia, pyridine, triethylamine, etc., and can be used again as a raw material in the method of the present invention.

Next, the present invention will be explained in more detail with reference to Examples, but the present invention is not limited to these Examples.

Examples 1 to 9 2.0 y of (R,S)-α-cyano-8-phenoxybenzylethyl carbonate and 40 ml of each esterase listed in Table 1 were added to 2M 11°C acetate buffer (PH 4,0).
15-, and the mixture was reacted with stirring at 40°C. After reacting for 24 hours, the reaction product was extracted with toluene.

The extract was subjected to high performance liquid chromatography (HP LC) [
Lichrosorb RP-13, methanol-water (
6:4), 254 planes, UV detection], α-cyano-3-phenoxybenzyl alcohol and α-cyano-8
- The hydrolysis rate was calculated from the peak area ratio with phenoxybenzylethyl carbonate.

After concentrating the extract, silica gel column chromatography was performed and cyclohexane-ethyl ether (95:
5) Elute with a solution to separate and remove unreacted α-cyano-8-phenoxybenzylethyl carbonate, and then elute with methanol containing a small amount of p-toluenesulfonic acid to remove free α-cyano. -3-phenoxybenzyl alcohol was obtained.

10~ of the obtained free α-cyano-8-phenoxybenzyl alcohol was dissolved in toluene 1- and equimolar amount of (S)-(t)-2-(4-chlorophenyl)-isovaleric acid chloride. Pyridine is added and reacted to produce (S)-← of α-cyano-3-phenoxybenzyl alcohol.
)-2-(4-chlorophenyl)-isovaleric acid diastereomer, and gas chromatography (column: D
Optical isomer analysis was performed using CQF-1,2,5m, column temperature = 2506).

The results are shown in Table 1.

/ Examples 9 to 14 Liquid medium in a 500 Md shoulder flask [For bacteria (Example 9.10), sweetened bouillon medium (1 t of water, glucose IO, OP, peptone 5.6 R, meat extract 5.Of,
Dissolve NaCl2.0 and adjust the pH to 7.2. ),
For frequent use of mold and yeast (Examples 11 to 14), malt extract, yeast extract medium (dissolve 5.0 g of peptone, 10.01 g of glucose, 8.0 g of malt extract, and 8.0 g of yeast extract in 1 t of water) , P)I shall be 6.5.)]1
After sterilization by adding 00d, two platinum loops of each microorganism listed in Table 2 were inoculated from the slant culture, and cultured with reciprocating shaking at 30°C for 72 hours.

Next, the pH of each culture solution was adjusted to 4.5 using a 2M HC/- aqueous solution, and (R,S)-α-cyano-8
-Phenoxybenzylpropyl carbonate3. Of was added, and the mixture was reacted at 30° C. for 30 hours with stirring. Thereafter, the reaction products were separated in the same manner as in Example 1, and the optical isomer type ratio and hydrolysis rate of the obtained α-cyano-3-phenoxybenzyl alcohol were measured.

The results are shown in Table 2.

(18 completed)

Claims (1)

[Claims] General formula (wherein k is an alkyl group having 1 to 8 carbon atoms;
It represents an alkenyl group having 1 to 8 carbon atoms or an alkynyl group having 1 to 8 carbon atoms. ) A microorganism having the ability to preferentially asymmetrically hydrolyze the carbonate ester of monolithic alcohol by acting on the carbonate ester of (R,S)-α-cyano-8-phenoxybenzyl alcohol shown in The esterase produced by 81
1 (R,S) p to the carbonate ester of alcohol
It is characterized by reacting at H7 or less and asymmetrically hydrolyzing it to split it into (S), an optically active α-cyano-8-phenoxybenzyl alcohol, and its enantiomer carbonate ester. Biochemical method for producing optically active α-cyano-8-phenoxybenzyl alcohol.