JPH0520072B2 - - Google Patents

Description

[Detailed description of the invention]

(Industrial Application Field) The present invention relates to a method for producing (R)-3-halogeno-1,2-propanediol. (R)-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. For example, (R)-3-chloro-1,2-propanediol is used in the synthesis of L-carnitine (Japanese Patent Application Laid-Open No. 165352/1983). (Prior art and problems) Regarding the production method of (R)-3-halogeno-1,2-propanediol, Hayden F. Jones describes methyl-5-chloro-5-deoxy-α-L-arabinofuranoside. How to obtain more information, [Chemistry and Indu-
stry), P.533, 15July, 1978], or H.
A method of obtaining it from 1,2,5,6-diacetonyl-D-mannitol by Jackson et al. [Chemistry and Biological Interactions (Chem.-Biol. Interactions) 13 , 193
(1976)], similarly the method of Y. Kawakami et al. [Journal of Organic Chemistry]
(Journal of Organic Chemistry) 47 , 3581
(1982)] are known. 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 discovered that (R)-3-halogeno-1,2-
As a result of intensive research aimed at improving the industrial properties of propanediol, we found that an inexpensive (R-S)-3-halogen-
1,2-propanediol(S)-3-halogeno-
By using microorganisms that have the ability to selectively metabolize 1,2-propanediol, (S)-
It has been discovered that 3-halogeno-1,2-propanediol can be selectively emitted and (R)-3-halogeno-1,2-propanediol can remain, and the present invention has been completed. Microorganisms that can be used in the present invention include the genus Nadsonia, the genus Saccharomyces, the genus Saccharomycopsis, the genus Rhodosporidium, the genus Rhodotorula, and the genus Wickerhamia.
hamia), Stephanoascus, Torulopsis, Pachysolen
Genus, Schizosaccharomyces, Sporobolomyces, Brettanomyces, Escherichia, Aeromonas
Genus, Acidiphilium,
Micrococcus spp., Bacillus spp., Enterobacter spp., Nocardia spp.
Genus, Protaminobacter
Genus, Pseudomonas, Arthrobacter, Hafnia, Klebsiella,
Microorganisms belonging to the genus Rhodococcus and Gluconobacter can be mentioned. For more information, please visit Nadsonia
Elongata (Nadsonia elongata) IFO 0665,
Saccharomyces cerevisiae
cerevisiae) IFO 0267, Saccharomycopsis lypolytica
IFO0717, Rhodosporidium toruloides IFO0871, Rhodotorula rubra
IFO0383, Wickerhamia fluorescens IFO1116, Stephanoascus
ciferrii) IFO1854, Torulopsis gropengiesseri
IFO0659, Pachsolen. tannophilus IFO1007, Schizosaccharomyces
pombe) IFO0362, Sporobolomyces salmonicolor
IAM12249, Brettanomyces custersianus IFO1585, Escherichia coli IFO12734,
Aeromonas hydrophila
hydrophila) IFO3820, Acidiphilium cryptum (Acidiphilium cryptum) IFO14242,
Micrococcus luteus (Micrococcus
luteus) IFO12708, Bacillus cereus (Bacillus cereus) IFO3001, Enterobacter aerogenes (Enterobacter aerogenes)
IFO13534, Nocardia globerula IFO13509, Protaminobacter alboflavus
alboflavus) IFO3707, Pseudomonas fragi IFO3458, Pseudomonas crucivi-ae
IFO12047, Pseudomonas chlororaphis IFO3904, Arthrobacter simplex
simplex) IFO12069, Hafnia alvei IFO3731, Klebsiella pneumonias IFO12009,
Rhodococcus erythropolis
erythropolis) IFO12320, Gluconobacter. Gluconobacter suboxydans
There are IFO3254, 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 used in normal culture, such as inorganic salts, trace metal salts, and vitamins, can be appropriately mixed and used. The above microorganisms may be cultured by conventional methods, for example, the culture medium pH should be in the range of 4.0 to 9.5, and the culture temperature should be in the range of 20 to 45.
It is preferable to culture aerobically at a temperature in the range of 10 to 96 hours. As a method for obtaining (R)-3-halogeno-1,2-propanediol by allowing microorganisms to act on (R,S)-3-halogeno-1,2-propanediol,
A method in which a substrate is added to the culture solution cultured as described above or a suspension of bacterial cells obtained from this culture solution by centrifugation, or a method in which a substrate is added to the medium and culture and reaction are carried out simultaneously, a conventional method. There is a method in which a substrate is added to a suspension of immobilized microorganisms in an appropriate buffer solution. Reaction temperature is 15~
It is preferable to carry out the reaction at 50°C and the reaction pH in the range of 4.0 to 10.0, and a buffer or the like can be used as appropriate to maintain the pH. The substrate concentration in the reaction solution is 0.1~
The amount is preferably 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, oxygen amount, 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. (R)-3-halogeno obtained in this way
To collect 1,2-propanediol from the reaction solution, 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 dehydrating the extract with anhydrous sodium sulfate, etc., and removing the solvent under reduced pressure, (R)
A syrup of -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 1-28 Glucose 4%, ( _NH4 ) _2HPO4 1.3% _{,
KE2PO4 0.7} %, _MgSO4・_7H2O800ppm ,
_ZnSO4・_7H2O60ppm , _FeSO4・_7H2O90ppm ,
_CuSO4・_5H2O5ppm , _MnSO4・_4H2O10ppm ,
A medium consisting of 100 ppm NaCl and 0.3% yeast extract was prepared with deionized water (PH 7.0), and 500 ml each was dispensed into two Sakaguchi flasks and sterilized at 120°C for 20 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. Sterilize 500 ml of the reaction solution by centrifugation, concentrate the supernatant to approximately 50 ml, and add 150 ml of ethyl acetate three times (total).
450ml) was extracted. 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 this product was measured, the values shown in Table 1 were obtained. [Literature value of (R)-3-chloro-1,2-propanediol [α] ²² _D -6.9 (c = 2H ₂ O)] In addition, after tosylating each syrup by a conventional method, the optical isomer Hexane-
HPLC using isopropyl alcohol (95:5) as a solvent at a flow rate of 2.0 ml/min and a wavelength of 235 nm.
Analysis (retention time: S form: 35 minutes, R form: 38.5 minutes) revealed that each sample contained the corresponding (R) form.

【table】

【table】

[Table] However, the yield was determined from the added (R,S)-3-chloro-1,2-propanediol. In addition, only in Example 28, a medium (PH7.0) consisting of 2% glucose, 2% glycerin, 2% corn steep liquor, 0.3% yeast extract, and 1% calcium carbonate was used, but other conditions were not used. Example 1~
I did the same thing as 27. Examples 29-56 Using the strains shown in Table 1, the substrate was (R,S)-3
Table 2
The results shown are obtained. [Literature value of (R)-3-bromo-1,2-propanediol [α] ²⁵ _D -3.94 (c=5.07CHCl ₃ )]

【table】

【table】

[Table] However, the yield was determined from the added (R,S)-3-bromo-1,2-propanediol, and was calculated from Example 56.
The medium of Example 28 was used for culturing.

Claims (1)

[Scope of Claims] 1. Nadsonia genus, Saccharomyces genus, Saccharomycopsis genus, Rhodos genus, which have the ability to selectively metabolize S-3-halogeno-1,2-propanediol. Rhodosporidium, Rhodotorula, Wickernamia, Stephanoascus, Torulopsis, Pachysolen
Genus, Schizosaccharomyces, Sporobolomyces, Brettanomyces, Escherichia, Aeromonas
Genus, Acidiphilium,
Micrococcus spp., Bacillus spp., Enterobacter spp., Nocardia spp.
Genus, Protaminobacter
Genus, Pseudomonas, Arthrobacter, Hafnia, Klebsiella,
Microorganisms belonging to the genus Rhodococcus and Gluconobacter are allowed to act on (R,S)-3-halogeno-1,2-propanediol, and the remaining R-3-halogeno-
A method for producing optically active (R)-3-halogeno-1,2-propanediol, which comprises collecting 1,2-propanediol. 2 The microorganisms are Nadsonia elongata, Saccharomyces cerevisiae, and Saccharomycopsis lipolyteica.
lipolytica), Rhodosporidium toruloides, Rhodotorula rubra, Uicerhamia
fluorescens (Wickerhamia fluorescens),
Stephanoascus ciferrii, Torulopsis gropengiesseri,
Pachysolen tannophyllus (Pachysolen)
tannophilus), Schizosaccharomyces pombe, Sporobolomyces salmonicolor
salmonicolor), Brettanomyces custersianus, Escherichia coli, Aeromonas hydrophila,
Acidiphilium cryptum, Micrococcus luteus, Bacillus cereus, Enterobacter aerogenes, Nocardia globerula, Protaminobacter alboflavus (Protaminobacter alboflavus), Pseudomonas fragi, Pseudomonas cruciviae,
Pseudomonas chlororaphis, Arthrobacter simplex,
Hafnia alvei, Klebsiella pneumonias,
Phodoccocus erythropolis
erythropolis) and Gluconobacter suboxydans. 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.