JP2761064B2 - Production method of optically active 1,3-butanediol - Google Patents

Description

The present invention relates to a method for producing optically active 1,3-butanediol. More specifically, an optically active 1,3-butanediol is obtained by allowing a specific microorganism or a processed product thereof to act on an enantiomeric mixture of 1,3-butanediol and collecting the remaining optically active 1,3-butanediol. It relates to a method for producing butanediol.

Optically active 1,3-butanediol is an important synthetic raw material for various pharmaceuticals such as antibiotics.

[Problems to be solved by conventional technology and invention]

Conventionally, methods for producing optically active 1,3-butanediol include (1) a method of producing a chemically synthesized racemic 1,3-butanediol.
A method of optically resolving butanediol using an optical resolving agent (JP-A-61-191631), and (2) 4-hydroxy-butanediol using a Raney nickel catalyst treated with an optically active compound.
Method for asymmetric synthesis from 2-butanone (JP-A-58-204187)
Bull. And Bull. Chem. Soc. Jpn., 53, 1356-1360 (198
0)) and the like are known. However, both (1) and (2) require the use of expensive optical resolving agents and catalysts,
(2) has disadvantages such as low optical purity, so it is economically excellent and has a high optical purity by a simple means.
It is desired to establish a method for obtaining 3-butanediol.

[Means for solving the problem]

The present inventors are economically excellent, and by a simple method, as a method for obtaining optically active 1,3-butanediol having high optical purity, focused on a method of acting a microorganism, and searched for a microorganism suitable for this purpose. , Aciculonidium (Acicul
oconidium), Brettanomyces
Genus, genus Cochliobolus, genus Corynespora, genus Dactylium, genus Echinopodospora, Hamigella
igera), Helminthosporiu
m) A microorganism selected from the group of microorganisms belonging to the genera, Nectria, and Fialocephal, acts on an enantiomeric mixture of 1,3-butanediol, leaving (R) -1,3-butanediol. Letting go, and Ambrosiozyma, Bordetella, Fusarium, Gibberella, Gonatobo
tryum), Neosartorya, Oospora, Paecilomyces
Genus, Preussia, Spetori
a) A microorganism selected from the group of microorganisms belonging to the genera, Talaromyces, and Westerdykella, acts on an enantiomeric mixture of 1,3-butanediol, and (S) -1,3-butane The inventors have found that diols remain in diols, and have completed the present invention.

Microorganisms used in the present invention include genus Aiculoconidium, Brettanomyces (Brett
anomyces), Cochliobolus, Corynespora, Dactyli
um), Echinopodospora, Hamigera, Helmin sosporium (Helmin)
thosporium), a bacterium selected from the group of genus Nectria, and a group of microorganisms belonging to the genus Phialocephala, which act on an enantiomeric mixture of 1,3-butanediol to produce (R) -1,3-butanediol. Microorganisms having the ability to remain, or genus Ambrosiozyma, genus Bordetella,
Fusarium genus, Gibberella
Genus, genus Gonatobotryum, genus Neosartorya, genus Oospora,
Paecilomyces, Preuss
ia), Spetoria, Talaromies (Ta
laromyces), Westerdykella
A microorganism selected from the group of microorganisms belonging to the genus, which acts on an enantiomeric mixture of 1,3-butanediol,
Any microorganism that has the ability to allow 1,3-butanediol to remain can be used.

Specifically, microorganisms that act on an enantiomeric mixture of 1,3-butanediol and have the ability to allow (R) -1,3-butanediol to remain, include Acycloconidium aculeatum IFO
10124, Brettanomyce anomaras
s anomalus) IFO 0796, Cochliobolus miyabeanus IFO 6631, Corynespora cassiicola IFO 6724,
Dactylium dendroide
s) ATCC 46032, Eshinopodospora jamaithesis (E
chinopodospora jamaicensis IFO 9819; Hamigera avellanea IFO 7721; Helminthosporium sigmoideum varity illegulare (Helminthosporium sigmoideum var.irregular)
e) IFO 5273, Nectria cinna
barina) IFO 6821, and Phyalocephala bactrospora IFO 8770.

Microorganisms that act on an enantiomeric mixture of 1,3-butanediol and have the ability to retain (S) -1,3-butanediol include Ambrosiozyma monospora IFO 1965 and Ambrosiozyma filamentant (Ambrosiozyma philentoma) I
FO 1847, Bordetella bronchiseptica (Bordetella)
bronchiseptica IFO 13691, Fusarium oxysporum IFO 7151, Fusarium soiani IFO 5232, Gibberella fujikuroi IFO 5268, Gonatobotorium apicratum goulabo
m) IFO 9098, Neosartorya fischeri var. spinosa
IFO 5955, Oospora
astringenes) IFO 7001, Pesilomies barioty (P
aecilomyces variotii) IFO 4855, Preussia terricola IFO 7893, Spetoria glycines IFO 5294, Talaromyces flavas Balaity flavus (Talaromyces fl
avus var. flavus) IFO 7231, Westerdykella multispora IFO 5813 and the like.

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, 8th edition, vol.
8) and can be obtained from the IFO. Microorganisms with ATCC numbers are American Type Cult
Catalog of Bacteria issued by ure Collection (ATCC)
Phages rDNA Vectors, 16th edition (1985) and can be obtained from the ATCC. Microorganisms with IAM numbers can be obtained from the Institute of Applied Microbiology, The University of Tokyo.

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. Culture at a temperature of 20 to 45 ° C., preferably 25 to 37 ° C., anaerobically or aerobically under conditions suitable for the growth of the microorganism for 5 to 120 hours, preferably about 12 to 72 hours. .

As a method for producing an optically active 1,3-butanediol from an enantiomer mixture of 1,3-butanediol, a method of adding an enantiomer mixture of 1,3-butanediol to the culture solution using the culture solution as it is, The cells are separated by centrifugation and the like, or
After washing, resuspend in buffer, water, etc.
There is a method in which an enantiomeric mixture of butanediol is added and reacted. At the time of this reaction, it may be better to add a carbon source such as glucose or sucrose as an energy source. The cells may be live cells, or may be cells that have been subjected to treatment such as crushed cells, acetone treatment, and freeze fitting. In addition, these cells or the treated cells are immobilized by a known method such as a polyacrylamide gel method, a sulfur-containing polysaccharide gel method (a carrageenan gel method, etc.), an alginic acid gel method, an agar gel method, and the like. You can also. Further, an enzyme purified and obtained from a treated product of the cells by a combination of known methods can be used.

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

The reaction is carried out at pH 3-10, preferably at pH 5-9 and at a temperature of 10-10.
The reaction is carried out at a temperature in the range of 6060 ° C., preferably 20-40 ° C., for about 1-120 hours under stirring or standing. When the reaction time is prolonged, the residual amount of 1,3-butanediol decreases, but it is possible to obtain optically active 1,3-butanediol having high optical purity. The concentration of the substrate used is not particularly limited, but may be 0.1
About 10% is preferable.

The optically active 1,3-butanediol remaining after the reaction can be easily collected by using a conventional purification method such as extraction with an organic solvent, distillation, column chromatography, etc., directly from the reaction solution or after isolation of cells.

〔Example〕

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

The amount of 1,3-butanediol in the reaction solution in the examples was determined by gas chromatography (column: Thermon 3).
000, (2m), temperature 130 ℃)
The optical purity was obtained by acetylating the optically active 1,3-butanediol obtained by the reaction with acetyl chloride by a conventional method,
High performance liquid chromatography using an optical resolution column (column: Chiral Cell OB manufactured by Daicel Chemical Industries, solvent: n-
Hexane / 2-propanol = 19: 1, wavelength 220 nm, flow rate 0.5
(retention time; (S) body 15 minutes,
(R) body 19.3 minutes).

Example 1 <Culture medium for preparation of microbial cells> 0.3% peptone 0.5% glucose 1.0% yeast extract 0.5% 1,3-butanediol _{K 2 HPO 4 0.1% MgSO 4} · 7H 2 O 0.05% pH7.2 made above bacteria health 100 ml of the culture medium was placed in a 500 ml Sakaguchi flask, sterilized, and inoculated with the microorganisms shown in Table 1, respectively.
Shaking culture was performed at 30 ° C. for 48 hours. Subsequently, the cells were separated by centrifugation and washed once with physiological saline to obtain viable cells.

Next, 50 ml of distilled water was placed in a 500 ml Sakaguchi flask, and the viable cells were suspended therein, and then 0.5 g of racemic 1,3-butanediol was added thereto, followed by a reciprocating shaking reaction at 30 ° C. for 48 hours. .

After the completion of the reaction, the bacteria were removed by centrifugation, and the obtained supernatant was saturated with sodium chloride, and then extracted with 50 ml of ethyl acetate.The ethyl acetate layer was analyzed by gas chromatography and remained. The amount of 1,3-butanediol present was measured.

Next, the ethyl acetate was dehydrated with anhydrous sodium sulfate and desolventized to obtain a syrup, which was acetylated with acetyl chloride in a conventional manner, dissolved in a solvent, and analyzed by high performance liquid chromatography to obtain a syrup. The absolute configuration and optical purity of 1,3-butanediol were measured.

 Table 1 shows the obtained results.

[Effect of the Invention] The method for producing an optically active 1,3-butanediol using the microorganism of the present invention can be easily performed using an optically active 1,3-butanediol having high optical purity.
This makes it possible to produce butanediol, which is extremely advantageous industrially.

Claims (2)

(57) [Claims] 1. The method according to claim 1, wherein the enantiomeric mixture of 1,3-butanediol is mixed with an acycloconidium.
Genus, genus Brettanomyces, genus Cochliobolus, Corynespor
a) genus, genus Dactylium, genus Echinopodospora, genus Hamigera,
Helminthosporium, Nectria, and Phialocephal
a) a microorganism selected from the group of microorganisms belonging to the genus, acting on a mixture of enantiomers of 1,3-butanediol and acting on a microorganism having the ability to leave (R) -1,3-butanediol or a processed product thereof; , The remaining optically active (R)-
A process for producing optically active 1,3-butanediol, comprising collecting 1,3-butanediol. 2. An enantiomeric mixture of 1,3-butanediol is added to the genus Ambrosiozyma, the genus Bordetella, the genus Fusarium, the genus Gibberella, and the gonatobotorium.
tryum), Neosartorya, Oospora, Paecilomyces
Genus, Preussia, Spetori
a) selected from the group of microorganisms belonging to the genera, genus Talaromyces, genus Westerdykella, acting on a mixture of enantiomers of 1,3-butanediol to produce (S) -1,3-butanediol; A process for producing optically active 1,3-butanediol, which comprises reacting a microorganism having a capability of remaining or a processed product thereof to collect the remaining optically active (S) -1,3-butanediol.