JPH0690789A - Production of @(3754/24)s)-1-phenyl-1,3-propanediol - Google Patents

Abstract

PURPOSE:To obtain the subject compound useful as raw material for synthesizing various medicines industrially and advantageously by treating an enantiomer mixture of 1-phenyl-1,3-propanediol with a bacterium or its treated material and collecting the residual compound. CONSTITUTION:A bacterium [e.g. Bacillus amyloliquefaciens (IFO14,141)] or its treated material is subjected to shaking culture in a medium containing an enantiomer mixture of 50% (R) derivative and (S) derivative of 1-phenyl-1,3- propanediol at 30 deg.C for 24-48 hours and further the enantiomer mixture of 50% (R) derivative and 50% (S) derivative of 1-phenyl-1,3-propanediol is added to the culture mixture and subjected to shaking culture at 30 deg.C for 72 hours. After the reaction, the reaction solution is diluted with ethanol, centrifuged to remove the bacteria, a supernatant liquid is subjected to high-performance liquid chromatography to simply obtain remaining (S) 1-phenyl-1,3-propanediol having high optical purity.

Description

Detailed Description of the Invention

[0001]

The present invention relates to (S) -1-phenyl-
The present invention relates to a method for producing 1,3-propanediol. (S)
-1-Phenyl-1,3-propanediol is useful as a raw material for the synthesis of various drugs.

[0002]

2. Description of the Related Art Conventionally, (S) -1-phenyl-1,3-
As a method for producing propanediol, a method of asymmetrically hydrolyzing ethyl 3-phenyl-3-hydroxypropionate with lipase and chemically reducing the obtained (S) -3-phenyl-3-hydroxypropionic acid (U.S. Pat. No. 4,921,798) or a method in which a microorganism is allowed to act on an enantiomeric mixture of 1-phenyl-1,3-propanediol to leave (S) -1-phenyl-1,3-propanediol remaining (Japanese Patent Laid-Open No. Hei 4 (1998) -43868). -58896, Japanese Patent Laid-Open No. Hei 4
-88999, JP-A-4-94691, JP-A-4-15
2896) is known. However, the former method has the drawback that the raw materials are expensive and the operation is complicated. In the latter method, the raw material is inexpensive, but depending on the microorganisms found to date, the optical purity is high (S) -1 in a short time.
There was a problem that -phenyl-1,3-propanediol could not be left.

[0003]

SUMMARY OF THE INVENTION The object of the present invention is to inexpensively and conveniently provide (S) -1-phenyl-1, which has a high optical purity,
It is to provide a method for producing 3-propanediol.

[0004]

DISCLOSURE OF THE INVENTION As a result of repeated studies in view of the above circumstances, the present inventors have found that (R) -1 preferentially acts on an enantiomer mixture of 1-phenyl-1,3-propanediol. It was found that there is a microorganism capable of decomposing phenyl-1,3-propanediol and, as a result, leaving (S) -1-phenyl-1,3-propanediol remaining, and completed the present invention. It was

That is, the present invention relates to 1-phenyl-1,
A culture of a microorganism having an ability to act on an enantiomer mixture of 3-propanediol and leave (S) -1-phenyl-1,3-propanediol remaining, a microbial cell isolated from the culture, or The treated product of the microbial cells is allowed to act on the enantiomeric mixture of 1-phenyl-1,3-propanediol, and the (S) -1-phenyl-1,3-propanediol produced is collected ( The present invention provides a method for producing S) -1-phenyl-1,3-propanediol.

The microorganism used in the present invention has the ability to act on an enantiomeric mixture of 1-phenyl-1,3-propanediol to leave (S) -1-phenyl-1,3-propanediol. Any microorganism may be used, but specific examples include the following. Bacillus amyloliquefaciens IFO14141 Brevibacterium luteum IFO12676 Nocardia restricta ATCC14887 Pseudomonas stutzere Hansenula anomala) IFO0145 Hansenula ciferrii IFO0905 Hansenula petersonii IFO1372 Kloeckera javaob 66 moths. FO1038 Prototheca Sophie (Prototheca zopfii) ATCC16527

These microorganisms can be used as any strain such as a wild strain, a mutant strain, or a recombinant strain derived by a genetic technique such as cell fusion or genetic engineering.

The medium for culturing the microorganism used in the present invention is not particularly limited as long as the microorganism can grow. For example, a normal liquid nutrient medium containing a carbon source, a nitrogen source, inorganic salts, organic nutrients and the like can be used.

As the carbon source, any of the above microorganisms can be used so long as it is available. Specifically, sugars such as glucose, fructose, sucrose and dextrin, alcohols such as sorbitol, ethanol and glycerol, and fumaric acid. , Organic acids such as citric acid, acetic acid and propionic acid or salts thereof, hydrocarbons such as paraffin and mixtures thereof can be used.

Examples of the nitrogen source include inorganic ammonium salts such as ammonium sulfate and ammonium chloride, organic acid ammonium salts such as ammonium fumarate and ammonium citrate, nitrate salts such as sodium nitrate and potassium nitrate, peptone, yeast extract and meat extract. An organic nitrogen compound such as corn steep liquor or a mixture thereof can be used.

In addition, nutrients such as inorganic salts, trace metal salts and vitamins which are usually used in culture can be appropriately mixed and used. If necessary, a factor that promotes the growth of microorganisms, a factor that enhances the ability to produce the target compound of the present invention, or a substance effective for maintaining the pH of the medium can be added.

As a culture method, pH is 3 to 9.
5, preferably 4 to 8, culture temperature is 20 to 4
It is cultivated at 5 ° C., preferably 25 to 37 ° C., anaerobically or aerobically under conditions suitable for the growth of the microorganism for 5 to 120 hours, preferably for 12 to 72 hours.

As the method for allowing the above-mentioned microorganism to act on the enantiomeric mixture of 1-phenyl-1,3-propanediol, the microorganism culture thus obtained is used as it is, or the bacterial cells are separated by centrifugation or the like, and this is left as it is. Alternatively, after washing, resuspended in a buffer solution, water, etc., a mixture of 1-phenyl-1,3-propanediol enantiomers may be added and reacted.
In this reaction, it may be better to add a carbon source such as glucose, fructose or sucrose as an energy source. The microbial cells may be intact microbial cells, or may be used as a microbial cell crushed product, an acetone-treated microbial cell, or a lyophilized microbial cell-treated product. In addition, these cells or treated cells can be used after being immobilized by a known method such as polyacrylamide gel method, carrageenan method, alginic acid gel method.

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

The reaction is pH 3 to 9, preferably pH 5
To 8 at a temperature of 10 to 60 ° C., preferably 20 to 40 ° C., with stirring or standing. The concentration of the substrate used is not particularly limited, but is 0.1 to 10
% Is preferable. Thus, by reacting for about 1 to 120 hours, the reaction solution has a high optical purity (S)-.
1-Phenyl-1,3-propanediol is produced and accumulated.

Collection of (S) -1-phenyl-1,3-propanediol produced by the reaction is carried out directly from the reaction solution or after separation of cells, followed by ordinary purification such as extraction with an organic solvent, distillation, column chromatography and the like. It can be easily obtained by using the method.

[0017]

EXAMPLES The present invention will now be described in more detail with reference to Examples. The absolute configuration, optical purity and reaction yield of 1-phenyl-1,3-propanediol in the examples were measured by high performance liquid chromatography (column: Daicel Chemical Industries, Chiralcel OB, eluent: hexane / 2-propanol). = 8/2, flow rate: 0.7 ml / min, detection:
UV 210 nm) (retention time: (S) form 8.75 minutes, (R) form 9.72 minutes).

Example 1 Glucose 2.0%, (NH ₄ ) ₂ SO ₄ 0.5%, K
₂ HPO ₄ 0.3%, KH ₂ PO ₄ 0.1%, MgSO
_{4 · 7H 2 00.05%, FeSO} 4 · 7H 2 O0.0
01%, MnSO ₄ .4H ₂ O 0.001%, yeast extract 1.0%, polypeptone 1.0% (pH)
7.0) was dispensed into test tubes in an amount of 5 ml, heat sterilized, and inoculated with a platinum loop amount of cells of the microorganisms shown in Table 1 obtained by culturing in broth agar medium at 30 ° C. for 24 hours in advance, The cells were shake-cultured at 30 ° C. for 24 to 48 hours. Then, 25 mg of an enantiomer mixture of 1-phenyl-1,3-propanediol containing 50% each of the (R) form and the (S) form was added to the culture solution, and the mixture was further shaken at 30 ° C. for 72 hours to react. went. After completion of the reaction, the reaction solution was diluted with ethanol and then sterilized by centrifugation, and the obtained supernatant was subjected to high performance liquid chromatography to remove residual 1-phenyl-
The absolute configuration of 1,3-propanediol, optical purity and reaction yield were measured. The results are shown in Table 1.

[0019]

[Table 1]

Example 2 1.0 l of the medium having the composition shown in Example 1 was dispensed into a 2 l volume mini-jar fermenter, heat-sterilized, and preliminarily shake-cultured in the same medium at 30 ° C. for 24 hours. Hansenula putter Inoculated with 80 ml of Sony IFO1372 culture solution, aeration rate of 1 vvm, stirring speed of 500 rpm, 30 ° C., 24
Culturing was carried out for an hour. After culturing, the cells are collected by centrifugation and the same amount of 200 mM phosphate buffer solution (pH
After washing with 7.0), the cells were suspended in 1.0 l of the same buffer solution. To this suspension, 5.0 g of the same enantiomeric mixture of 1-phenyl-1,3-propanediol used in Example 1 was added, and the reaction was carried out at 30 ° C. with aeration and stirring for 72 hours. After the reaction was completed, the remaining 1-phenyl-
When the absolute configuration of 1,3-propanediol, the optical purity and the reaction yield were measured, the absolute configuration was the (S) form, the optical purity was 100%, and the reaction yield was 27%.

Next, the reaction solution was filtered through an ultrafiltration membrane, and the obtained filtrate was mixed with toluene-isopropyl alcohol 1
Extraction was performed twice with 1 and the mixture was concentrated under reduced pressure, crystallized and dried to obtain 1.1 g of (S) -1-phenyl-1,3-propanediol crystals.

According to the method of the present invention, (S) -1-phenyl-1,3-propanediol having high optical purity can be easily produced.

─────────────────────────────────────────────────── ─── Continuation of the front page (51) Int.Cl. ⁵ Identification code Internal reference number FI technical display location (C12P 41/00 C12R 1: 365) (C12P 41/00 C12R 1:38) (C12P 41/00 (C12R 1:01) (C12P 41/00 C12R 1:78) (C12P 41/00 C12R 1: 645)

Claims (2)

[Claims] 1. A microorganism or a treated product thereof is allowed to act on an enantiomeric mixture of 1-phenyl-1,3-propanediol to collect the remaining (S) -1-phenyl-1,3-propanediol. (S) -1
-Method for producing phenyl-1,3-propanediol. 2. A microorganism selected from the group of microorganisms belonging to the genus Bacillus, Brevibacterium, Nocardia, Pseudomonas, Clavispora, Hansenula, Croquera, Rhodosporodium, Sporoboromyces, and Prototheca. The microorganism having the ability to act on an enantiomeric mixture of 1-phenyl-1,3-propanediol to leave (S) -1-phenyl-1,3-propanediol remaining. S) -Method for producing 1-phenyl-1,3-propanediol.