JPH0488999A - Production of optically active 1,3-propanediol - Google Patents

Abstract

PURPOSE:To simply obtain the title high-purity compound useful as an intermediate for producing drugs and agricultural chemicals by bringing a culture solution of microorganism into contact with a 1,3-propanediol of racemic modification and raising the ratio of S isomer. CONSTITUTION:A compound shown by the formula (X is H, halogen, OH, nitro or 1-5C alkyl) is subjected to contact reaction with cells (e.g. Candida tropicalis ATCC 20,006 or Saccharomyces SAKE KYOKAI no. 7), a treated material of cell or a culture solution to give the objective optically active 1,3-propanediol. Concentration of the substrate shown by the formula added is 0.01-4wt.% and reaction is carried out in a medium such as buffer solution or methanol preferably at 25-40 deg.C at pH6-9 preferably up to a point of time when optical purity becomes >=80% e.e.

Description

Detailed Description of the Invention (Field of Industrial Application) The present invention relates to a method for producing optically active 1,3-propanediols useful as intermediates for the production of pharmaceuticals and agricultural chemicals. Optical activity by increasing the abundance ratio of the three forms by allowing microorganisms to act on propanediol 1.
The present invention relates to a method for producing 3-propanediols.

(Prior art) As a method for producing optically active 1,3-propanediols, a method of reducing optically active 2,3-epoxy cinnamyl alcohol with Red-AI (J, Org, chem
, 53.4081 (1988)) and optically active 3-
A method in which hydroxy-3-phenylpropionic acid is esterified with diazomethane and then reduced with NaBHn [T
etrahedron Letters, Vol, 2
6. Nn3. p351 (1985)) and a method of optical resolution using liquid chromatography using a polysaccharide derivative as a carrier (Japanese Unexamined Patent Publication No. 61-176538). The above methods have drawbacks such as the production of by-products that are difficult to separate and purify, such as 1,2-diol, the use of highly toxic reagents, and the optical purity of the resulting products not necessarily high.

(Problems to be Solved by the Invention) The present invention aims to convert racemic 1,3-propanediol to S-1 in a simple process using microorganisms and under mild conditions.
, 3-propanediol.

(Means for Solving the Problems) As a result of extensive research to achieve the above object, the present inventors have discovered that by bringing specific microorganisms into contact with racemic 1,3-propanediol, five microorganisms The present inventors have discovered that the abundance ratio of S-1,3-propanediol is significantly increased, and as a result, S-1,3-propanediol with high optical purity can be obtained, and the present invention has been completed.

That is, the present invention provides racemic 1,3- Optical activity characterized in that S-1,3-propanediols are obtained by contacting propanediols with a culture solution, bacterial cells, or treated bacterial cells of a specific microorganism to increase the abundance ratio of 5-propanediols. The present invention provides a method for manufacturing a body.

The microorganisms used in the present invention include Candida, Satucharomyces, Pihia, Torulopsis, Debaryomyces, Rhodotorula, Pletanomyces, Roderomyces, Mucor, Cordyceps, Bracheslia, Coanephora, Rhizopus, and Alternaria. microorganisms belonging to the genera Fusarium, Penicillium, Trichoderma, Aspergillus, Cladosporium, Roselinia, Corynebacterium, and Rhodococcus.

Specific bacterial strains are illustrated below.

Candida tropicalis 8TCC20006, Candida parabsilosis IF00585, Satucharomyces salmon association no. 7, Pichia miso IFO0193
, Torulopsis xylinus IFO0454, Debaryomyces hanzenii IFO0564, Rhodotorula sp. ATCC20254, Pletanomyces anomalous IFO0642, Rhoderomyces elongisporus IFO1676, Plaquesria trisbora NRRL
2895, Coanephora trisbora NRRL 59
89, Penicillium chrysogenum IFO4626, Trichoderma viride IP04847, Corynebacterium nitrilophilus ATCC21419, Rhodococcus sp. AK 32 (Feikoken Bacillus No. 8269)
, Fusarium oxyborum JAM 5009゜AK3
The two strains are those described in JP-A No. 62-91189.

The microorganisms used in the present invention can be cultured according to known methods. The medium to be used can be one known as a nutrient source for general microorganisms, such as carbon sources such as glucose, ethanol, glycerin, and sucrose, nitrogen sources such as ammonium sulfate or urea, yeast extract, malt extract, peptone, and meat extract. organic nutrient source, phosphoric acid,
Inorganic nutrient sources such as magnesium, potassium, iron, and manganese can be used in appropriate combinations. In addition, 1°3
-Propanediols may be added. The pH of the medium should be selected in the range of 5 to 10 (the culture temperature should be 18 to 45
℃, preferably 27-37°C. The number of culture days is 1~
It may be cultured within 10 days until the activity reaches its maximum.

As a method for producing S-1,3-propanediols,
A method in which racemic 1゜3-propanediol represented by general formula (1) is added to the medium and the culture is continued, or a method in which the bacterial cells or bacterial cells grown by culture are collected in advance and the racemic cells are added to the medium and cultured is continued. There is a method of contacting the body with 1,3-propanediol (1). Examples of the processed bacterial cells include crushed bacterial cells, organic solvent-treated bacterial cells, and enzymes separated and extracted from the bacterial cells. As the reaction medium, an aqueous medium such as water or a buffer solution, a homogeneous mixed medium of a water-soluble organic solvent such as methanol or dimethyl sulfoxide and an aqueous medium, or a two-phase medium such as water-chloroform can be used. The racemic substrate represented by general formula (1) is added as it is or as a solution in an appropriate solvent. The concentration of the substrate added may range from 0.01 to 40% by weight. Reaction temperature: 5 to 50°C, preferably 25 to 40°C, reaction pH
is 4-11, preferably 6-9. The reaction is S-1,
The process may be stopped when the optical purity of 3-propanediol reaches a desired purity or higher. Preferably 80%e.

It is 80 or more.

The reaction mechanism of the present invention was studied. The microorganism used in the present invention has a weak activity of degrading the compound of formula (1). In addition, the number of moles of the 3 forms produced relative to the charged racemic form (1) (hereinafter referred to as the reaction rate) usually exceeds 50%, indicating that the R form in the racemic form has been converted to 3 forms. It can be said. Furthermore, when analyzing the time course of the reaction, the following equation (
2) It was found that prochiral β-hydroxyketones represented by the formula (wherein X is the same as above) were produced. Therefore, when we synthesized 3-hydroxypropiophenone as an example of β-hydroxyketone and exposed it to microorganisms, we found that 5-1-phenyl-
1,3-propanediol was produced. From the above, the present invention aims at converting the R form in the racemic 1,3-propanediol represented by the formula (1) to the compound of the formula (2), prochiralizing it, and then reducing it specifically to S. It was found that -1,3-propanediols were produced.

Specifically, the racemic 1,3-propanediols (1) used in the present invention include, for example, racemic 1-phenyl-1,3-propanediol and its benzene ring, including halogen, hydroxyl group, nitro Examples include compounds into which substituents such as groups and alkyl groups are introduced. As the halogen, for example, fluorine, chlorine, iodine, bromine, etc. are preferable.
The alkyl group is preferably a methyl group or an ethyl group having 1 to 5 ring carbon atoms.

The three 1,3-propanediols (1) produced according to the present invention can be easily purified by extraction, column purification, etc. after removing insoluble substances such as bacterial cells from the reaction-completed liquid.

In addition, three purified 1,3-propanediols (1
) was determined by IR and NMR, and also by HP
Optical purity can be determined by LC analysis and specific rotation measurement.

(Effects of the Invention) According to the present invention, optically active 1,3-propanediols useful as intermediates for the production of pharmaceuticals and agricultural chemicals can be produced with extremely high purity under reaction conditions of room temperature and normal pressure.

(Examples) Hereinafter, the present invention will be described in detail based on Examples, but the present invention is not limited thereto.

Example 1 Glucose 2.0%, yeast extract 0.3%, diammonium phosphate 1.3%, monopotassium phosphate 0.2%, sodium chloride 0.01%, sulfur M ferrous 0.01% , 1-
Candida balapsilosis ATCC2 cultured in advance in 1000 d of sterile medium containing 0.01% phenyl-1,3-propanediol and having a pH of 7.0.
0008 was inoculated at 2% and cultured at 30°C for 48 hours.

After culturing, the bacterial cells were collected by centrifugation and suspended in an Erlenmeyer flask containing 97 sdi of water.
-Phenyl-1,3-propanediol (500 μl) was added and reacted at 30°C for 24 hours. After the reaction was completed, the bacterial cells were removed by centrifugation, the supernatant was extracted with chloroform, and then purified using a silica gel column.
-Phenyl-1,3-propanediol 412 ml was obtained.

Melting point: 63.5-65°C Specific rotation: [α)”=-129.5° (C=1.0
, methanol, 365na+) Further, the optical purity was determined by the following HPLC analysis and was found to be 99.0% e, e.

Analysis conditions Column: C old RALCEL OB 25cmX0.4
6 cm (manufactured by Daicel Chemical Industries) Elution solvent: n-hexane: isopropanol 9:1 Flow rate: 0.6 d/sin Detection: UV 254 nm Example 2 In the same manner as Example 1, culture in sterilized medium 100- in advance in the same medium 2% of the microorganisms obtained were inoculated and cultured at 30°C for 60 hours. After culturing, the bacterial cells were collected by centrifugation, suspended in an Erlenmeyer flask containing 10M1 phosphate buffer, added with 20μ of racemic 1-phenyl-1,3-propanediol, and incubated for 24 hours. The reaction was carried out at °C. After the reaction is complete,
The reaction rate and optical purity of 5-1-phenyl-1,3-propanediol were determined by HPLC analysis. Table 1 shows the results.
Shown below.

2 parts of 3-hydroxypropiophenone were added to 101d, and the reaction was carried out at 32°C for 24 hours. Generated 5-1
The production rate and optical purity of -phenyl-1,3-propanediol were determined by HPLC. The results are shown in Table 2.

Table 2 (1 other person) Example 3

Claims (1)

[Claims] General formula (1) ▲ Numerical formulas, chemical formulas, tables, etc. ▼ (1) (In the formula, X is a hydrogen atom, a halogen atom, a hydroxyl group, a nitro group, or an alkyl group having 1 to 5 carbon atoms. Optical activity 1,3, characterized in that the abundance ratio of S isomer is increased by contacting racemic 1,3-propanediol shown in -Production method of propanediols.