JPH0469999B2 - - Google Patents

Description

[Detailed description of the invention]

(Industrial Application Field) The present invention relates to the optically active β-
This invention relates to a method for producing optically active epichlorohydrin using DCH as a raw material. (Prior Art) Optically active epichlorohydrin is an important raw material for the synthesis of various medicines. However, the method for producing this optically active epichlorohydrin is
Baldwin, Journal. of. Organic. Chemistry (J.Org.Chem) Volume 43, 1978, No.
Page 4876 or Ellis, Journal. chemical.
Society. , chemical communication. ,
(J.CHEM.SC., CHEM.COMMUN.) 1984,
Although they are described on page 1600, all of them require advanced synthesis techniques, and no simple manufacturing method is known. (Problem to be solved by the invention) The present inventor has already identified racemic β-DCH and R-(+)
- A method for obtaining highly pure optically active S-(-)-β-DCH by contacting it with β-DCH-assimilating bacteria (JP-A-Sho
61-132196) and a method to obtain R-(-)-epichlorohydrin by reacting this product with an alkaline agent (Japanese Patent Application Laid-open No. 62-6697), but the optical isomer opposite to these was developed. , that is, S-
A simple method for producing (+)-epichlorohydrin is not known. The present invention has solved this problem. (Means for Solving the Problems) The present inventors have completed the present invention by discovering that the above optically active epichlorohydrin can be easily produced with high purity by microbial treatment. That is, the present invention provides R-(+)- obtained by allowing bacteria belonging to the genus Alcaligenes or cultured cells thereof having the ability to assimilate S-(-)-β-DCH to react with racemic β-DCH in a medium. This is a method for producing optically active epichlorohydrin, which is characterized by reacting β-DCH with an alkaline agent to obtain S-(+)-epichlorohydrin. The mycological properties of the microorganisms isolated and collected from soil by the present inventor and used in the present invention are shown in Table 1.

【table】

[Table] Based on the above results, Bergey's Manual of Systematic Bacteriology
Based on the description in Volume 1 of Bacteriology, this bacterium has characteristics of the genus Alcaligenes. The present inventor named this bacterium Alcaligenes Sp. DS-K-S38 (hereinafter, this bacterium is referred to as DS-K-S38 strain). This bacterium was submitted to the Institute of Microbial Technology, Agency of Industrial Science and Technology.
It has been deposited as No. 11114 (FERM P-11114). In the present invention, not only the above-mentioned DS-K-S38 strain, its variants and mutants, but also any bacteria belonging to the genus Alcaligenes and having the ability to assimilate S-(-)-2,3-dichloro-1-propanol can be used. All can be used. The present invention provides the racemic β-
The main point is to optically activate DCH and then react with an alkaline agent using a conventional method to obtain optically active epichlorohydrin. In the present invention, the above-mentioned bacteria or their cultured cells may be used, or they may be immobilized. However, the methods for culturing and immobilizing the above-mentioned bacteria may be commonly used methods. That is, the culture method involves culturing the above-mentioned bacteria in a nutrient medium containing a carbon source, a nitrogen source, an organic nutrient source, and an inorganic nutrient source, such as a bouillon medium or a sweetened bouillon medium, allowing them to grow well, and culturing the resulting culture. Alternatively, cultured bacterial cells may be used. Carbon sources include carbohydrates such as glycerin, or organic acids such as citric acid, maleic acid, and malic acid and their salts; nitrogen sources include inorganic nitrogen such as ammonium sulfate, ammonium chloride, ammonium nitrate, and ammonium phosphate, and peptone. Organic nitrogen such as casein, yeast extract, meat extract, etc. can be used. Other inorganic salts include phosphates, magnesium salts, potassium salts,
Iron salts, zinc salts, copper salts, etc. are used. The culture conditions are usually about 20-45°C, preferably 25-37°C.
The reaction is carried out aerobically at a temperature of about 5° C. and a pH of about 5 to 9, preferably 6.0 to 7.5, by means of shaking or aeration. In addition, the immobilization method may be, for example, acrylamide, k
- A method of enclosing live bacterial cells using carrageenan, agar, gelatin, sodium alginate, etc. is sufficient;
After immobilization, it may be used by crushing it into an appropriate size and shape. The reaction between the above-mentioned bacteria and racemic β-DCH can be carried out by stirring the above-mentioned bacteria, their cultured cells, or their immobilized products in a medium containing racemic β-DCH, such as a synthetic medium, and bringing them into good contact. The contact time is usually half a day to 10 days, and the concentration of β-DCH in the medium may be about 0.1 to 0.6% by volume. After the reaction is completed, the reaction solution is taken out and filtered to separate the cultured cells and the supernatant, or the immobilized material and the supernatant, and the R-(+)-β-DCH remaining in the supernatant is removed. It is fractionated by operations such as activated carbon column treatment, ether extraction, and vacuum distillation. This fraction is treated with an alkaline agent, preferably a caustic alkali such as caustic soda or caustic potash, to produce epichlorohydrin. Furthermore, if immobilized bacterial cells are used in the method of the present invention, operations such as centrifugation become easy, and the immobilized product can be used repeatedly. (Example) The present invention will be specifically explained below using examples. In the examples, percentages are by weight unless otherwise specified. Example 1 Medium 20 containing yeast extract 1.0%, glycerin 2.0%, polypeptone 1.0%, pH 7.0 was placed in a 30 jar fermenter, and after sterilization by heating in the usual manner, DS-
K-S38 strain was inoculated and cultured for 24 hours under the following conditions. Temperature: 30°C PH: Initial pH: 7.0 Aeration rate: 20/min Stirring speed: 300 rpm After the cultivation was completed, the microbial cells and the culture filtrate were separated using a centrifuge to obtain 600 g of viable bacterial cells. continue,
Suspend the viable bacterial cells in the synthetic medium shown below10.
After adjusting the volume, it was fixed with acrylamide in a conventional manner. The immobilized product was crushed into pieces of 0.5 to 1 mm square using a mixer and thoroughly washed with a synthetic medium. Components of synthetic medium Ammonium sulfate 0.05% by weight Ammonium nitrate 0.05 〃 Potassium hydrogen phosphate 0.1 〃 Sodium phosphate 0.2 〃 Disodium phosphate 0.1 〃 Magnesium sulfate 0.05 〃 Iron sulfate, copper sulfate, manganese sulfate Trace amounts PH Initial PH6. 8 Next, the immobilized product prepared in this way 100
Place in a jar fermenter and adjust to 80 ml with synthetic medium. Furthermore, racemic β-
320 ml of DCH and 160 g of calcium carbonate were added and stirred under the following conditions. Temperature: 30℃ Aeration rate: 40/min Rotation speed: 300r.pm 72 hours after the start of the reaction, the supernatant liquid and immobilized material were filtered, and the remaining β-DCH was extracted from this liquid using an activated carbon column, ether extraction, and vacuum distillation. Sort and separate
152g was collected. The substance was identified using the following method. 1) Identification by gas chromatography As a result of comparison with commercially available β-DCH using column carrier PEG-20MP, 5%, 60-80 mesh,
The retention times were exactly the same. Purity 98.2%
that's all. 2) Identification by IR (infrared absorption spectrum) As shown in the chart shown in Figure 1, its absorption pattern was exactly the same as that of commercially available β-DCH. From the above, this substance was clearly found to be β-DCH. The following method was used to confirm that this substance was R-(+)-β-DCH. 1) Measurement of optical rotation The specific optical rotations of commercially available β-DCH and this substance are as follows. Commercially available β-DCH [α] ²⁰ _D = 0.0゜ C = 1, dichloromethane actual substance [α] ²⁰ _D = +10.4゜ C = 1, dichloromethane 2) R-(+)-α-methoxy-α-trifluoro Preparation of methyl phenyl acetate ester and analysis by high performance liquid chromatography R-(+)-α-methoxy-α-trifluoromethyl phenyl acetate chloride was commercially available β
After reacting with -DCH and this substance to prepare its ester derivative, the analysis results by liquid chromatography were as follows. Analysis conditions Column carrier ZORBAXODS 4.6mm x 25cm (manufactured by Du Pont) Eluent Methanol:water = 65:35 (V/V) Elution volume 1ml/min Detection method Absorbance analysis results at 260nm Commercially available β-DCH Retention times 50.5 and 52.0 Two peaks with the same area were given in minutes. This substance gives a peak only at a retention time of 52.0 minutes.
No peak was observed at 50.5 minutes. 3) Preparation of dichloropropyl-N-phenyl carbamate and its optical rotation Commercially available β-DCH, 1 g of this substance, and 0.9 g of phenyl isocyanate were added to 30 ml of dry acetone and 0.3 ml of triethylamine, and heated under reflux for about 3 hours.
After preparing the dichloropropyl-N-phenyl carbamate, its specific rotation was measured. Commercially available β-DCH [α] ²⁵ _D = 0.0゜ C = 1, methanol This substance [α] ²⁵ _D = +16.4゜ C = 1, methanol From the above results, this substance is R-(+)-β-
It was found that it was DCH and its optical purity was over 99%. Next, 100 g of this R-(+)-β-DCH was mixed with 650 ml of 1.4N caustic soda aqueous solution in a 1000 ml flask, stirred vigorously at room temperature for 80 minutes, and then ether was added.
After adding 200 ml and stirring, the ether layer was separated. Subsequently, the ether layer was dried with magnesium sulfate, and then
Ether was distilled off, and epichlorohydrin was further distilled to obtain 60.3 g. The purity of this epichlorohydrin was 99.4% or more as measured by gas chromatography. Further, the specific optical rotation was as follows. [α] ²² _D = +34.3° (C = 3.4, methanol) That is, the obtained epichlorohydrin is S-
It was (+)-epichlorohydrin, and its optical purity was 99% or more. Example 2 As in Example 1, culture medium 2 containing 1.0% yeast extract, 1.0% polypeptone, 2.0% glycerin, and pH 7.0 was placed in a 5-volume jar fermenter, and as usual.
After heat sterilization, DS-K-S38 strain was inoculated, and Example 1
Cultured for 24 hours under the same conditions. Next, Example 1 was placed in a 100-volume jar fermenter.
80g of synthetic medium shown in and 160g of calcium carbonate,
Add 320 ml of racemic β-DCH and 40 g of polypeptone, heat sterilize, inoculate the above culture as usual, temperature 30℃, aeration rate 40/min, rotation speed.
The reaction was performed while culturing at 300 rpm. 48 hours after the start of the reaction, the reaction solution was separated into supernatant, bacterial cells, and precipitate using a centrifuge, and the remaining β-DCH was collected from the supernatant in the same manner as in Example 1. , 148 g of R-(+)-β-DCH was obtained. The specific optical rotation of the obtained R-(+)-β-DCH was [α] ²⁰ _D = +10.4° (C = 1.0, dichloromethane), and as a result of analysis in the same manner as in Example 1, the optical purity was
It was over 99%. Further, the conversion of R-(+)-β-DCH to S-(+)-epichlorohydrin was carried out in the same manner as in Example 1, and the specific optical rotation [α] ²² _D
=+34.3° (C=3.4, methanol) S-(+)-epichlorohydrin with an optical purity of 99% or more was obtained. (Effects of the Invention) According to the present invention, 2,3-dichloro-
Optically active S-(+)-epichlorohydrin can be obtained more easily and with higher purity than 1-propanol via optically active R-(+)-2,3-dichloro-1-propanol.

[Brief explanation of the drawing]

FIG. 1 shows infrared absorption spectra of R-(+)-2,3-dichloro-1-propanol, a raw material of the present invention, obtained in Example 1 and a commercially available product of the same substance. - is commercially available β-DCH, --- is R-
(+)-β-DCH is shown.

Claims (1)

[Scope of Claims] 1. Bacteria belonging to the genus Alcaligenes having the ability to assimilate S-(-)-2,3-dichloro-1-propanol, or cultured cells thereof, are treated in a culture medium with racemic 2,
An optical system characterized by reacting R-(+)-2,3-dichloro-1-propanol obtained by reacting with 3-dichloro-1-propanol with an alkali agent to obtain S-(+)-epichlorohydrin. Process for producing active epichlorohydrin. 2. The production method according to claim 1, wherein bacteria belonging to the genus Alcaligenes having the ability to assimilate S-(-)-2,3-dichloro-1-propanol or cultured cells thereof are used after being immobilized.