JPS59130190A - 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 or derived from animal organs. CONSTITUTION:The esterase produced by a microbial strain capable of conducting preferential asymmetric hydrolysis of the ester of (S)-alcohol when made to contact with the ester of (R, S)-alpha-cyano-3-phenoxybenzyl alcohol of formula (A is 1-4C halogen-substituted alkyl, alkenyl or alkynyl), e.g. Arthrobacter simplex IFO 3530, or the esterase derived from animal organs, is made to contact with the ester of said (R, S)-compound at <=7pH, preferably 3.5-6.5pH to resolve the racemic mixture into the ester of an optically active alpha-cyano-3-phenoxybenzyl alcohol rich in (S)-isomer and the ester of its antipode.

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to a biochemical process for the production of optically active α-cyano-3-phenoxybenzyl alcohol.

More specifically, general formula [I] N [wherein A is a halogen-substituted alkyl group having 1 to 4 carbon atoms,
It represents a halogen-substituted alkenyl group having 1 to 4 carbon atoms or a halogen-substituted alkynyl group having 1 to 4 carbon atoms. ] (R,S) -(t-cyano-3-fe/-
4 Acting on the ester of dibenzyl alcohol, (S)
An esterase produced by a microorganism having the ability to preferentially asymmetrically hydrolyze an ester of -α-cyano-3-phenoxybenzyl alcohol or an esterase derived from an animal organ is represented by the above general formula [I] (RtS) An ester of monoalcohol is reacted at pH 7 or lower, and it is hydrolyzed (divided into optically active α-cyano-3-phenoxyben 1.dyl alcohol rich in Sl monomer and its enantiomer ester). The present invention relates to a biochemical method for producing optically active α-cyano-3-phenoxybenzyl alcohol.

σ-Cyano-3-phenoxybenzyl alcohol is known as a new 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 its response (Kosuke Yoshioka, Organic Synthetic Chemistry, Vol. 38, No. 12,
1151-1162 (1980)) Therefore, (g,s)-α-cyano-3-phenoxybenzyl alcohol obtained by ordinary synthetic chemical production methods was optically resolved by an industrially advantageous method to obtain ( It has been desired to develop a technology to obtain Sl integrally.

However, since α-cyano-3-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 optical activity. Currently, reagents are required.

The present inventors have developed an industrially advantageous production method for vine (S)-α
- As a result of repeated research to find a method for producing cyano-3-phenoxybenzyl alcohol, we found that the racemic form represented by the general formula CI), that is, the ester of (R,s)-α-cyano-3-phenoxybenzyl alcohol, was used as a raw material. By biochemically asymmetric hydrolysis of Toshi and Kore, optically active α-cyano-3-phenoxybenzyl alcohol rich in (S) monomer and its enantiomer (D They found that it can be divided efficiently, and after further various studies, they have completed the present invention.

That is, the present invention provides (R,S) represented by the general formula [1]
- Esterase produced by a microorganism that has the ability to preferentially asymmetrically hydrolyze the ester of (S)-α-cyano-3-phenoxybenzyl alcohol by acting on the ester of α-cyano-3-phenoxybenzyl alcohol ( In the present invention, esterase refers to esterase in a broad sense including lipase) or esterase derived from animal organs, represented by the above general formula [I] (R2S)
Let it act on the ester of alcohol at pH 7 or less,
Optically active α-cyano-3-phenoxybenzyl alcohol is obtained by hydrolyzing this and dividing it into optically active α-cyano-3-phenoxybenzyl alcohol, which is rich in (S), and its enantiomer ester. Provide manufacturing method.

Next, the manufacturing method of the present invention will be described in detail.

In the present invention, (R) is used as a raw material.

Esters of S)-α-cyano-3-phenoxybenzyl alcohol can be prepared by conventional methods for the preparation of esters, e.g. (R,S)
-α-cyano-3-phenoxybenzyl alcohol has the general formula [■] AC-OH[]T] 1 [wherein A has the same meaning as above. It can be easily produced by reacting a carboxylic acid halide or acid anhydride represented by the following formula. Alternatively, it can be easily produced by reacting 3-phenoxybenzaldehyde, sodium cyanide, and a carboxylic acid halide represented by the above general formula [■].

The carboxylic acid represented by the above general formula CII) is preferably a □ carboxylic acid having 1 to 2 halogen atoms at the α-position, particularly monochloroacetic acid or monobromoacetic acid, which has 1 to 1 at the α-position. More preferred are carboxylic acids having 6 chlorine or bromine atoms.

The microorganism that produces the esterase used in the present invention is (Rts)-α represented by the general formula [T].
-Any microorganism that produces an esterase that has the ability to act on esters of cyano-3-phenoxybenzyl alcohol and preferentially asymmetrically hydrolyze esters of monolithic alcohol (Sl-(he)) can be used regardless of its species. Can be used without any problem.

Examples of such microorganisms include Arthrobacter (Arth).
robacter), Pseudomona7. (Ps
eudomonas), ri0-fE:Haku7-!
7 A (Cbromobacterium), Bacillus (Bacillus), Candida (Ca)
ndida) genus, Nocardia (Nocar d
i a) Genus Rhodotorula
) Genus, Brevi-b
acterium) genus, Trichobium/l/-6, (
Trichoderma ) genus, Rhizopus (Rhizo
(Mucor) genus, Aspergillus (Aspergillus)
Genus, Geotricum, Hough *
5 < Ra (Hansenulli genus, Enterobacter
(Enterobacter) genus, Flavobacterium (Flavobacterium) genus, Mycobacterium (Mycobacterium) genus, Corynebacterium (Corynebacterium) genus,
Lactobacill u
Genus, Streptomyces
) genus, Penicillium genus,
Actinomucor: genus Actinomucor, genus Gibberella, genus Absidia, Cu
genus Gliocladium (
Gliocla old um) genus, Saccaromyces (Sacc)
haro-myces), Cryptococcus (Cr
yp tococcus), Pich 7
ia) genus, Micrococcus ((us)
Genus, Torulopsis, Aureobasidium,
Alcaligenes
Genus or Achromobacter (Achromobacter)
r) microorganisms belonging to the genus. Among these,
In the production method of the present invention, hydrolysis rate and optical selectivity), Arthrobacter sp., Achromobacter sp., Pseudomonas sp., Chromobacterium sp., Bacillus sp., Brevibacterium sp., Nocardia sp., Rhodotorula sp. , Candida spp., Trichoderma spp., Rhizopus spp.
Microorganisms belonging to the genera Mucor, Aspergillus, Geotrichum, Aureobasidium, Hansenula, and Torulopsis are particularly useful.

Specific examples of this microorganism include the following bacterial cells.

(1) 7 Ruthrobacter simplex
IFO-3530 Arthrobacter si
complex (2) Achromobacter genus
ATCC-219ATCC-21910Ach
ro S P .

(3+ Pseudomonas 0 fluorescens
IFO-3081Pseudomonas flu
orescens (4) Chromobacterium viscosum ATCC-6918ChATCC-6
918Chro viscosum+5) Hafrus licheniformis IFO-12197
Bacillus IicheniformisBr
evibacterium arrmoniage
nes(7) Nocardia erythropolis
IFO-1232ONocardia ery
thropolis (8) Rhodotorula minuta burr texensis IFO-0879Rhodo
torula m1nuta var, texens
is(9) Candida utilis
IFσ-1086Candida uLilis oo) Trichoderma viride IFO-
4847Trichoderma virideα1
) Rhizopus chinensis IFO-4737R
hizopus chinensis02) Mucor javanicus IFO-4572Mu
cor j avan ic u 5Q31
Aspergillus li←ru◇Huspel IFO
-s324Aspergillus vading Oa
sper04 Geotrichum candium
IFO-5368 Geotricum can
didum05) Aureobasidium pullulans IFO-4464Aureobasi
dium pullulans06) Hansenula anomala IFO-0707Hanse
Torulopsis candida IF
O-0380Torulopsis can id
a All of these strains were purchased from the Fermentation Research Institute (IFO) or American Type Cu.
1ture Collection (ATCC) l
(It has been preserved and is available at this preservation institution.

In addition, some esterases derived from microorganisms and animal organs 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 asymmetric hydrolysis of the ester of (R,S)-α-cyano-3-phenoxybenzyl alcohol represented by the general formula CI) is performed using a culture solution in which the microorganism is cultured, or a culture solution. This is carried out by mixing an aqueous suspension of bacterial cells or a treatment product thereof, such as an aqueous solution containing crude esterase or purified esterase, with the ester of the (R2S) monoalcohol, and stirring or shaking the mixture. At this time, a non-ester surfactant may be added if necessary, and it is also possible to immobilize the enzyme.

Further, the reaction temperature is suitably 10 to 65°C, particularly preferably 20 to 50°C. The reaction time is usually 30 minutes to 24 hours.

It is important that the pH during the reaction is below pH 7.
,5 to pH 5.5. The concentration of the ester substrate used is in the range of 1 to 80 φ1, preferably 5 to 35% v/w, based on the reaction solution.

After carrying out the asymmetric hydrolysis reaction in this way, the reaction solution is subjected to operations such as static separation and solvent extraction to remove the free optically active α-cyano-3-phenoxybenzyl alcohol and the unreacted The enantiomer ester is separated and recovered. For this separation and recovery, operations such as solvent extraction and column chromatography can be employed as appropriate.

For example, by extracting the reaction solution with an organic solvent such as ether, benzene, or toluene, and subjecting this extract to column chromatography using silica gel, etc., and eluting with a mixture of dichlorohexane and ether (95:5), Free optically active α-cyano-3-phenoxybenzyl alcohol and unreacted ester of its enantiomer can be separated and obtained.

The unreacted 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 for 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 4.0 μl of monochloroacetic ester of (R,S)-α-cyano-3-phenoxybenzyl alcohol and 40 μl of each esterase listed in Table 1 below were added to a 0.2 M acetate buffer ( pH 4,0) 15-, and the mixture was reacted at 40° C. with stirring.

During the reaction, as the reaction progressed, an aqueous NaOH solution with an IM concentration was made into minute water droplets using a drop controller, and the pH was controlled to be constant using a pH controller while carefully adding the solution. After reacting for 5 hours, the reaction product was extracted with toluene. The extract was subjected to high performance liquid chromatography (HPLC).
) (Lichrosorb RP-18, methanol-water (6:4), 254 nm 5 UV detection), the peaks of α-cyano-3-phenoxybenzyl alcohol with monochloroacetic acid nistert and α-cyano-3-phenoxybenzyl alcohol were detected. The hydrolysis rate was calculated from the area ratio.

The extract was concentrated, subjected to silica gel column chromatography, and dichlorohexane-ethyl ether (95:5)
After elution with a solution of unreacted l-cyano-3-phenoxybenzyl alcohol, further elution with methanol containing a trace amount (io-5%) of no-radruenesulfonic acid removed the free cyano-3-phenoxybenzyl alcohol. 10 μm of 3-phenoxybenzyl alcohol was dissolved in 1 d of toluene, and 1 mole of this (5 μm) was dissolved in 1 d of toluene.
1-(4)l-2-(4-chlorophenyl)-isovaleric acid chloride and pyridine were added and reacted to form α-cyano-3-phenoxybenzyl alcohol (Sl-(+
)-2'(4-chlorophenyl)-isovaleric acid diastereomer and gas chromatography (column: 1
0% DCQF-1, 2.5 pain, column temperature: 250°C
) was used for optical isomer analysis.

The results are shown in Table 1.

Examples 10 to 14 500 - In a flask with a shoulder, put a liquid medium [malt extract, yeast extract medium (1.e of water, 5.02 peptone, 1 glucose)
0. Dissolve OP, 3.0 g of malt extract, and 3.0 g of yeast extract, and adjust the pH to 5.5. )] After sterilization, 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 pH 4.5 using 2Mali HCJ aqueous solution, and (R+S)-α-cyano-3
-2.0 mL of monobromoacetic ester of phenoxybenzyl alcohol was added, and the mixture was reacted at 30° C. for 15 hours with stirring. During the reaction, the pH was adjusted in the same manner as in Examples 1 to 9.
was controlled at a constant level. Thereafter, the reactants were separated in the same manner as in Examples 1 to 9, 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.

(22 completed)

Claims (3)

[Claims] (1) General formula [I] [wherein A is a halogen-substituted alkyl group having 1 to 4 carbon atoms,
It represents a halogen-substituted alkenyl group having 1 to 4 carbon atoms or a halogen-substituted alkynyl group having 1 to 4 carbon atoms. ] Acting on the ester of (k,S)-α-cyano-3-phenoxybenzyl alcohol represented by fsl-α-
An esterase produced by a microorganism having the ability to preferentially asymmetrically hydrolyze an ester of cyano-3-phenoxybenzyl alcohol or an esterase derived from an animal organ is used to synthesize the (R,S) monolithic alcohol represented by the general formula (II). An optical method characterized by reacting an ester at a pH of 7 or lower and hydrolyzing it to separate it into optically active α-cyano-3-phenoxybenzyl alcohol rich in (S) monomer and its enantiomer ester. Active α-cyano-3
- Biochemical production method of phenoxybenzyl alcohol. (2) (R9S)-α represented by the general formula [■]
- In the ester of cyano-3-phenoxybenzyl alcohol, the substituent A is a halogen-substituted alkyl group having 1 to 4 carbon atoms, having 1 to 2 halogen atoms at the bonding site, or a halogen-substituted carbon number 1-4 The manufacturing method according to claim 1, which represents an alkenyl group or a halogen-substituted alkynyl group having 1 to 4 carbon atoms. (3) (RIS)-α represented by the above general formula (I)
- In the ester of cyano-3-phenoxybenzyl alcohol, the substituent A is a halogen-substituted alkyl group having 1 to 4 carbon atoms or a halogen-substituted alkenyl group having 1 to 4 carbon atoms, in which the substituent A has one halogen atom at its bonding site. or a halogen-substituted alkynyl group having 1 to 4 carbon atoms, the manufacturing method according to claim 1 or 2.