JPH0521557B2 - - Google Patents

Description

[Detailed description of the invention]

(Industrial Application Field) The present invention relates to a method for producing (S)-3-halogeno-1,2-propanediol. (S)-3-halogeno-1,2-propanediol is a substance useful as a raw material for the synthesis of various pharmaceuticals and optically active physiologically active substances. Especially (S)-3-chloro-1,2
- Propanediol is known as a substance that has antifertility effects on sperm of various animals [H. Jackson et al., Chemical and Biological Interactions] 17 , 117 (1977)]. (Prior art and problems) Regarding the production method of (S)-3-halogeno-1,2-propanediol, the method of obtaining it from methyl-6-chloro-6-deoxy-α-D-glucopyranoside by HFJones (West German patent No. 2743858)
Method for obtaining 1,2,5,6-diacetone-D-mannitol by Porter KE et al. [Chem.-
Biol. Interactions 41 , 95 (1982)]. However, these methods cannot be called industrial production methods because the raw materials are difficult to obtain and the steps are complicated. A method for producing propanediol has been desired. (Means and Effects for Solving the Problems) The present inventors have discovered that (S)-3-halogeno-1, which has been conventionally obtained through complicated reactions using expensive raw materials,
As a result of intensive research aimed at industrial production of 2-propanediol, (R)-3-halogeno-1 was produced by allowing microorganisms to act on inexpensive (R,S)-3-halogeno-1,2-propanediol. ,2-
selectively metabolizes propanediol, (S)-3
-Halogeno-1,2-propanediol was found to remain, and the present invention was completed. Examples of microorganisms that can be used in the present invention include the genus Corynebacterium and the genus Proteus. For more information, see Corynebacterium acetoacidophilum.
ATCC 21476, Proteus morganii IFO 3168, etc. As the medium composition for culturing the above-mentioned microorganisms, any medium in which these microorganisms can grow can be used. For example, sugars such as glucose, sucrose, and maltose as carbon sources,
Alcohols such as ethanol, glycerol, and 1,2-propanediol, organic acids such as acetic acid and lactic acid, or mixtures thereof; nitrogen sources such as ammonium sulfate, ammonium phosphate, urea, yeast extract, meat extract, and peptone; Nutrient sources commonly used in culture, such as salts, trace metal salts, and vitamins, can be appropriately mixed and used. The above-mentioned microorganisms may be cultured by a conventional method, for example, it is preferable to culture the microorganism aerobically for 10 to 96 hours at a medium pH of 4.0 to 9.5 and a culture temperature of 20 to 45°C. As a method for obtaining (S)-3-halogeno-1,2-propanediol by allowing microorganisms to act on (R,S)-3-halogeno-1,2-propanediol, the culture solution cultured as described above or this A method in which a substrate is added to a suspension of bacterial cells obtained from a culture solution by centrifugation, a method in which a substrate is added to a medium and culture and reaction are performed simultaneously, a method in which microorganisms immobilized by a conventional method are added to an appropriate buffer There is a method of adding a substrate to a suspension in a liquid. The reaction temperature is
It is preferable to carry out the reaction at a temperature of 15 to 50°C and a reaction pH of 4.0 to 10.0, and a buffer or the like may be used as appropriate to maintain the pH. The substrate concentration in the reaction solution is
The amount is preferably 0.1 to 10 (W/V)%, and the substrate may be added all at once at the beginning of the reaction or may be added in portions.
The reaction is usually carried out with shaking or stirring, and the reaction time varies depending on the substrate concentration, the amount of enzyme, and other reaction conditions, but it is preferable to select conditions so that the reaction is completed in 24 to 120 hours. In terms of yield, it is preferable to stop the reaction by analyzing the remaining substrate by gas chromatography or the like, and stopping the reaction when the remaining substrate reaches approximately 50% of the added substrate. In order to collect the thus obtained (S)-3-halogeno-1,2-propanediol from the reaction solution, a general (R,S)-3-halogeno-1,
A method for collecting 2-propanediol can be used. For example, after removing bacterial cells from the reaction solution by centrifugation or the like, the supernatant is appropriately concentrated and extracted with a solvent such as ethyl acetate. After the extract is dehydrated with anhydrous sodium sulfate and the solvent is removed under reduced pressure, a syrup of (S)-3-halogeno-1,2-propanediol can be obtained. Further, it may be further purified by distillation. (Example) The present invention will be specifically described below with reference to Examples. Examples 1, 2 Glucose 4%, (NH ₄ ) ₂ HPO ₄ 1.8%, KH _2
PO4 0.7%, _MgSO4・_7H2O 800ppm,
ZnSO ₄・7H ₂ O 60ppm, FeSO ₄・7H ₂ O
90ppm, CuSO ₄・5H ₂ O 5ppm, MnSO ₄・4H ₂
O 10ppm, NaCl 100ppm, yeast extract 0.3
A medium consisting of % was made with deionized water (PH7.0),
2. Dispense 500ml each into Sakaguchi flasks and incubate at 120℃ for 20 minutes.
Sterilized for minutes. Inoculate each of the bacterial strains shown in Table 1 into the above medium, and
Shaking culture was carried out at ℃ for 48 hours to obtain culture solution 1.5. This culture solution is centrifuged to collect the bacterial cells, and after washing with water, the bacterial cells are added to 500 ml of 0.3M phosphate buffer (PH7.0).
(R,S)-3-chloro-1,2
- 2g of propanediol was added and reacted at 30°C for 72 hours with shaking. 500 ml of the reaction solution was sterilized by centrifugation, the supernatant was concentrated to about 50 ml, and extracted three times (450 ml in total) with 150 ml of ethyl acetate. The extract was dehydrated with anhydrous sodium sulfate and the solvent was removed under reduced pressure to obtain a syrup. When the specific optical rotation of each was measured, the values shown in Table 1 were obtained. [Literature value of (S)-3-chloro-1,2-propanediol [α] ²⁰ D + 7.3 (c = 1 H ₂ O) In addition, after tosylating these syrups by a conventional method, optical isomers Separation column [Chiralcel OC column (0.46 cm x 25 cm)] (manufactured by JASCO Corporation) using a solvent of hexane-isopropyl alcohol (95:5) at a flow rate of 2.0 ml/min and a wavelength of 235 nm.
As a result of HPLC analysis (retention time: S form: 35 minutes, R form: 38.5 minutes), it was confirmed that the corresponding (S) forms were contained.

[Table] However, the yield was determined from the added (R,S)-3-chloro-1,2-propanediol. Examples 3 and 4 Using the same strains as in Table 1, the substrate was (RS)-3-
Table 2
The results shown are obtained.

【table】

Claims (1)

[Scope of Claims] 1 A microorganism belonging to the genus Corynebacterium or the genus Proteus that has the ability to selectively metabolize (R)-3-halogeno-1,2-propanediol (R,S)-3- Optically active (S)-3-halogeno-1, characterized in that it acts on halogeno-1,2-propanediol and collects the remaining (S)-3-halogeno-1,2-propanediol;
Method for producing 2-propanediol. 2 The microorganism is Corynebacterium acetoacidophyllum (Corynebacterium acetoacidophyllum).
philum), Proteus morganii (Proteus morganii)
morganii). 3 (R,S)-3-halogeno-1,2-propanediol is (R,S)-3-chloro-1,2-
The manufacturing method according to claim 1 or 2, which is propanediol. 4 (R,S)-3-halogeno-1,2-propanediol is (R,S)-3-bromo-1,2-
The manufacturing method according to claim 1 or 2, which is propanediol.