JP2828742B2 - Method for producing optically active 3-phenyl-1,3-propanediol - Google Patents

Description

The present invention relates to a method for producing optically active 3-phenyl-1,3-propanediol. More specifically, enantiomeric mixtures of 3-phenyl-1,3-propanediol include:
The present invention relates to a method for producing optically active 3-phenyl-1,3-propanediol, which comprises reacting a microorganism or a processed product thereof and collecting remaining optically active 3-phenyl-1,3-propanediol.

Optically active 3-phenyl-1,3-propanediol is an important synthetic intermediate for various pharmaceuticals.

[Problems to be solved by conventional technology and invention]

Conventionally, as a method for producing optically active 3-phenyl-1,3-propanediol, a regioselective chemical reduction method of 2,3-epoxycinnamyl alcohol (J. Org. Ch.
em., 53 (17), 4081 (1988) and a method by the scientific reduction of optically active 3-phenyl-3-hydroxypropionic acid (Tetrahederon Lett., 26 (3), 351 (1985)). However, the former method has insufficient regioselectivity and insufficient chemical purity, and the latter method requires that the optically active organic acid must be resolved with an optical resolving agent. There are drawbacks such as low optical purity, etc. Under these circumstances, economically excellent and optically active 3-phenyl-1,3,3 with high optical purity by simple means.
-It is desired to establish a method for obtaining propanediol.

[Means for solving the problem]

The present inventors have focused on a method of allowing a microorganism to act as a method for obtaining optically active 3-phenyl-1,3-propanediol having high optical purity by an economically excellent and simple method, and a microorganism suitable for this purpose. Found that microorganisms selected from the group of microorganisms belonging to the genus Candida, the genus Hansenula, or the genus Rhodotorula act on an enantiomeric mixture of 3-phenyl-1,3-propanediol (R) -3-phenyl-
Leave 1,3-propanediol and the genus Candida, Geotrichum
A microorganism selected from the group of microorganisms belonging to the genera, genus Leucosporidium, genus Pichia, genus Pichia, genus Torulaspora, or genus Trichosporon is 3-phenyl-1,3-propanediol. Acting on an enantiomeric mixture, (S) -3-
The inventors have found that phenyl-1,3-propanediol remains, and have completed the present invention.

Microorganisms used in the present invention include Candida (Ca
ndida), a microorganism belonging to the genus Hansenula (Hansenula), or the genus Rhodotorula (Rodotorula);
A microorganism having the ability to act on an enantiomeric mixture of 3-propanediol and capable of remaining (R) -3-phenyl-1,3-propanediol, or a genus of Candida, a genus of Geotrichum, or a leucopoly Genus Leucosporidium, Pichia
ia) a microorganism belonging to the genus Genus, Torulaspora or Trichosporon, which acts on an enantiomeric mixture of 3-phenyl-1,3-propanediol and (S) -3-phenyl-1,3-propane Any microorganism can be used as long as it has the ability to leave the diol.

Specifically, microorganisms which act on an enantiomeric mixture of 3-phenyl-1,3-propanediol and are capable of remaining (R) -3-phenyl-1,3-propanediol include Candida lambica. (Candida la
mbica) DSM 70090, Hansenula minuta
nuta) DSM 70274, Rodotorula rubra
bra) AHU 3945, AHU 3948 and the like.

It also acts on an enantiomeric mixture of 3-phenyl-1,3-propanediol to produce (S) -3-phenyl-1,
Fine materials having the ability to leave 3-propanediol include candy pintropesii va pintropesii (Candida pintolopesii ver. Pintolopesi).
i) IFO 0729, Geotricum Candi Dam
m candidum) IFO 4597, IFO 4598, IFO 5368, IFO 31810, J
CM 1747, JCM 5222, Geotricum Fermentans (Ge
otrichum fermentans) JCM 2467, JCM 2468, Geotrichum klebahnii JCM 2171, Leucosporidium scoti
ttii) IFO 1923, Pichia querc
uum) DSM 70386, Torraspora del Brukki
laspora delbrueckii) IFO 0955 and Trichosporon capitatum IFO 1197.

Any of these microorganisms can be suitably used, such as a wild-type strain, a mutant strain, or a recombinant strain derived by a genetic technique such as cell fusion or genetic engineering.

Microorganisms with IFO numbers are described in List of Cultures, published by the Fermentation Research Institute (IFO), 8th edition, vol.
8) and can be obtained from the IFO. Microorganisms with AHU numbers can be found in the Catalog of Cultures, 4th edition (198
It is described in 7) and can be obtained from the Faculty of Agriculture, Hokkaido University. Microorganisms with JCM numbers are described in the microorganism strain Catalok, 4th edition (1989) issued by RIKEN Microorganism Preservation Facility, and can be obtained from the facility. Microorganisms with DSM numbers are Deatsche Sammlu
Catalog of stra published by ng von Mikroorganismen (DSM)
ins (1989) and can be obtained from the DSM.

The medium for culturing the microorganism used in the present invention is not particularly limited as long as the microorganism can grow. For example,
As the carbon source, any of the above microorganisms can be used, and specifically, glucose, fructose,
Sugars such as sucrose and dextrin, sorbitol,
Alcohols such as ethanol and glycerol, organic acids and salts thereof such as fumaric acid, citric acid, acetic acid, and propionic acid, hydrocarbons such as paraffin, and mixtures thereof can be used. As a nitrogen source, for example,
Ammonium chloride, ammonium sulfate, ammonium salts of inorganic acids such as ammonium phosphate, ammonium fumarate, ammonium salts of organic acids such as ammonium citrate, meat extract, yeast extract, corn steep liquor,
Casein hydrolysates, inorganic and organic nitrogen-containing compounds such as urea,
Alternatively, mixtures thereof can be used. In addition, nutrients used in ordinary culture, such as inorganic salts, trace metal salts, and vitamins, can be appropriately mixed and used. If necessary, a factor that promotes the growth of the microorganism, a factor that enhances the ability to produce the target compound of the present invention, or a substance that is effective in maintaining the pH of the medium can be added.

As a culture method, the medium pH is 3.0 to 9.5, preferably 4 to
8. The cultivation temperature is 20-45 ° C, preferably 25-37 ° C, under anaerobic or aerobic conditions suitable for the growth of the microorganism.
The culture is performed for about 120 hours, preferably about 12 to 72 hours.

As a method for producing optically active 3-phenyl-1,3-propanediol from an enantiomer mixture of 3-phenyl-1,3-propanediol, a culture solution is used as it is, and 3-phenyl-1,3 is added to the culture solution. -A method of adding an enantiomeric mixture of propanediol, separating cells by centrifugation, etc., washing the cells as they are or washing them, and then adding 3-phenyl-1,
There is a method in which an enantiomer mixture of 3-propanediol is added and reacted. During this reaction, glucose,
In some cases, it is better to add a carbon source such as sucrose as an energy source. The cells may be living cells, or may be those obtained by treating cells such as crushed cells, acetone treatment, freeze-drying and the like. In addition, these cells or treated cells, for example, polyacrylamide gel method,
It can be immobilized and used by a known method such as a sulfur-containing polysaccharide gel method (such as a carrageenan gel method), an alginic acid gel method, and an agar gel method. Further, an enzyme purified and obtained from a treated product of the cells by a combination of known methods can also be used.

The enantiomer mixture of 3-phenyl-1,3-propanediol can be reacted as it is, dissolved in water, or dissolved in an organic solvent that does not affect the reaction, or dispersed in a surfactant or the like. It may be added all at once or dividedly from the beginning.

The reaction is in the range of pH 3-10, preferably pH 5-9, and the temperature is
The reaction is carried out at 10 to 60 ° C, preferably 20 to 40 ° C, for about 1 to 120 hours under stirring or standing. When the reaction time is prolonged, the residual amount of 3-phenyl-1,3-propanediol often decreases, but it is possible to obtain optically active 3-phenyl-1,3-propanediol having high optical purity. The concentration of the substrate used is not particularly limited, but 0.1-20%
The degree is preferred.

Optically active 3-phenyl-1, which is produced by the reaction,
The collection of 3-propanediol can be easily obtained by using an ordinary purification method such as extraction with an organic solvent, distillation, column chromatography, etc., directly or after separating cells from the reaction solution.

〔Example〕

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

In the examples, 3-phenyl-1,3 in the reaction solution was used.
-The quantification and optical purity of propanediol were determined by directly subjecting the optically active 3-phenyl-1,3-propanediol obtained by the reaction to high-performance liquid chromatography using an optical resolution column (column: Daicel Chemical Industries Chiral Cell OB, solvent : n-hexane / 2-propanol = 19/1, wavelength 254 nm,
(Retention time: (S) 13.1 minutes, (R) 16.4) (flow rate: 1.0 ml / min, column temperature: 40 ° C., injection volume: 10 μ).
Minutes).

Example 1 <Cell culture medium> Glucose 2.0% Yeast extract 0.3% Peptone 0.5% Malt extract 0.3% pH 6.0 50 ml of the above cell culture medium was placed in a 500 ml Sakaguchi flask and sterilized. Each of the microorganisms thus obtained was inoculated, and cultured with shaking at 30 ° C. for 48 hours.

Next, after the completion of the culture, 0.25 g of a racemic 3-phenyl-1,3-propanediol was added, and a reciprocal shaking reaction was performed at 30 ° C. for 96 hours.

After completion of the reaction, the bacteria were removed by centrifugation, and the obtained supernatant was extracted with 50 ml of ethyl acetate, and the ethyl acetate was dehydrated with anhydrous sodium sulfate and desolvated to obtain a syrup.

This syrup was dissolved in 50 ml of a hexane / isopropyl alcohol (50/50) solution, and the amount, absolute configuration and optical purity of 3-phenyl-1,3-propanediol obtained by high performance liquid chromatography were measured. did.

 Table 1 shows the obtained results.

[Effect of the Invention] Optically active 3-phenyl-1,3 using the microorganism of the present invention
The method for producing propanediol makes it possible to easily produce optically active 3-phenyl-1,3-propanediol having high optical purity, and is extremely industrially advantageous.

──────────────────────────────────────────────────続 き Continued on the front page (51) Int.Cl. ⁶ Identification code FI (C12P 41/00 C12R 1:84) (C12P 41/00 C12R 1:78)

Claims (2)

(57) [Claims] 1. An enantiomeric mixture of 3-phenyl-1,3-propanediol is added to Candida
Genus, genus Hansenula or Rodtorla
otorula), a microorganism capable of acting on an enantiomeric mixture of 3-phenyl-1,3-propanediol to leave (R) -3-phenyl-1,3-propanediol. Alternatively, the optically active substance 3 is obtained by reacting the treated substance and collecting the remaining (R) -3-phenyl-1,3-propanediol.
A method for producing phenyl-1,3-propanediol. 2. An enantiomeric mixture of 3-phenyl-1,3-propanediol, wherein Candida
Genus, Geotrichum, Leucosporidium, Pichia, Torulaspora, or Tricosporone (Tri)
a microorganism selected from the group of microorganisms belonging to the genus chosporon and having the ability to act on an enantiomeric mixture of 3-phenyl-1,3-propanediol to leave (S) -3-phenyl-1,3-propanediol. Alternatively, the treated product is allowed to act, and the remaining (S) -3-phenyl-1,3-
A process for producing optically active 3-phenyl-1,3-propanediol, comprising collecting propanediol.