JPS62122597A - Production of (s)-3-halogeno-1,2-propanediol - Google Patents

Abstract

PURPOSE:To obtain the titled diol economically on an industrial scale, by treating (R,S)-3-halogeno-1,2-propanediol with a specific microorganism and collecting residual (R)-3-halogeno-1,2-propanediol. CONSTITUTION:(R,S)-3-halogeno-1,2-propanediol is treated with a microorganism belonging to Endomyces genus, Sporidiobolus genus, Geotrichum genus, Hansenula genus, Lodderomyces genus, Kluyveromyces genus, Nadsonia genus, Pichia genus, Saccharomyces genus, Saccharomycopsis genus, Rhodosporidium genus, Rhodotorula genus, Wickerhamia genus, Candida genus, Stephanoascus genus, Torulopsis genus, Pachysolen genus, Schizosaccharomyces genus, Sporobolomyces genus, etc., and remaining (R)-3-halogeno-1,2-propanediol is collected.

Description

DETAILED DESCRIPTION OF THE INVENTION (Industrial Field of Application) 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 L
-Used for carnitine synthesis (Japanese Patent Application Laid-Open No. 57-16
5352).

(Prior art and problems) (Regarding the method for producing R1-3-halogeno-1,2-propanediol, Hayden F. Jones described it from methyl-5-chloro-5-deoxy-α-L-arabinofuranoside. How to obtain, 1 Chemistry and Indus.
-stry), P, 533.15July, 197
8), and by H. Jackson et al. 1,2,
5. Method for obtaining 6-diacedonyl-D-mannitol [Chemistry and Biological Interactions (Chem, -Biol, Inter
actions) 13.

193 (1976)], also Y., Kawakam
I et al.'s method [Journal of Organic Chemistry (Journal of Organic Ch.
emistry) 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 manufacturing method was desired.

(Means and Effects for Solving the Problems) The present inventors have realized the industrial production of (R)-3-halogeno-1,2-propanediol, which has conventionally been obtained through complicated reactions using expensive raw materials. As a result of intensive research aimed at quality, we have developed an inexpensive (R,
By allowing microorganisms to act on S)-8-halogeno-1,2 propanediol, (S)-a-halogeno-1
, 2-propanediol is selectively metabolized, (R)-
The present inventors have discovered a scale in which 8-halogeno-1,2-propanediol remains, and have completed the present invention.

Microorganisms that can be used in the present invention include Entomyces (En
domyces), Sporideiobolus (Spo-r
idiobolus), Geotrichum (Geo tr)
ichum) genus, Hansenula (Hansenula)
Genus, Lodderomyces, Klu-yveromyces,
Genus Nadsonia, Pi
Chia) genus, Saccharomyces (Saccharomyces)
my c e s ) genus, Saccharomycopsis (Sac
char-omycops is), Rhodo-sporidium, Rhodotorula, Uikahamia (
Wickerhamia), Candida
ida), Stephanoascus (Stephanoascus)
ascus), Torulopsis
), Pachysolen genus, Schizosaccharomyces
s), genus Sporobolomyces
ces) genus, Brettanomyces
es) genus, Escher-ichia
Genus Aeromonas, Genus Acidiphilium, Genus Micrococcus, Genus Bacillus
aci-11us), Enterobacter (Enterobacter)
(obacter) genus, Nocardia (Nocardia)
) genus, Protaminobacter
ter) genus, Pseudomonas
), genus Arthur-obacter
), Hafnia genus, Klebsiella (K
levsiella), Rhodococcus (Rhodo-
coccus), Gluconobacter (Glucono
Examples include microorganisms belonging to the genus Bac-ter.

For more information, see Endomes Magnusi (Endo-m)
yces magnusii) CBS 164.
32. Sporidiobolus zijonsoniii (Spori
diobolus johnsonii) I F 0
6908, Geotricum candum (Geotr)
ichum candidum) CB5187, 67
, Hans-enula a
nomala) IF 0 0707, Lodderomyces elongysporus (Lodderomyces el)
ongisporus) I F O1676, Kluyveromyces fragilis (Kluyveromyce
s fragilis) IAM 476B, Nadsonia elongata
a) I F 0 0665, Pichia Bretoni (P
ichia burtonii) I F 00844
, Saccaromyces cerevisiae (Sacc-harom
yces cerevisiae) I F 0 02
67, Satucharomycopsis lipolytica (Sa cc
ha rom-ycopsis Iypolytica
) IF 0 0717, Rhodosporidium toru
loides) I F 0 0871, Rodo Torla・
Rhodotorula rubra I
F 00383, Ikahamya Fluorescence (
Wickerhami afluorescens)
I F O1116, Candeida humicola (C
andida humicola) CB 52774
, Cand-idaut
ilis) IFO0626, Stephanoascus ciferr
-1i) IFo 1854, Torulopsis groppengi
esseri) IFO0659, Pachsolen jannophilus
) I F 0 1007, Schizosacchar -omyces pombe
ombe) I F OO862, Sporobolomyces
Salmonica color (Sporobolomyces sa)
-lmonicolor) I AM 12249,
Pretanomyces custersianus (Brettano)
myces cus -tersianus) I F
O1585, Escherichia coli
hia coli) I F O12734, Aeromonas hy-
dropophila) I F O8820, Acidiphilium c
ryptum) IFO14242, Micrococcus luteus (Micrococcus 1uteus) I
F 0 12708, Bacillus cereus
I FO3001, Enterobacter aerogenes
aerogenes) I F O18584, Nocardia globela (Nocardia globela)
r-ula) IFo 13509, Protaminobacter alboflavus
albofla-vus) IFO8707, Pseudomonas fragi
) IF 0 8458, Pseudomonas cru-civiae
) IFO12047, Pseudomonas chlororaphi
s) IFO3904, Arthrobacter simplex I
F 012069, Hafni Albay
a al-vei) IFO3781, Klebsiella pneumonia
as) IF 012009, R-hodococcus erythrophori x
I F 0 12320, Glucon-obacter
er 5uboxydans) I F 0 3254
and so on.

As for the medium composition for culturing the microorganisms mentioned above, any medium in which these microorganisms can grow can be used. For example, carbon sources include glucose, sucrose, maltose natonosaccharides, ethanol, chrycerol,
1. Alcohols such as 2-7' lopanediol, organic acids such as acetic acid and lactic acid, or mixtures thereof, ammonium sulfate, ammonium phosphate, urea, yeast extract, meat extract, peptone, etc. as a nitrogen source, and inorganic salts,
Nutrient sources used in normal culture, such as trace metal salts and vitamins, can be appropriately mixed and used.

The above-mentioned microorganisms may be cultured by conventional methods, for example, the culture medium pH
is in the range of 4.0 to 9.5, and the culture temperature is 20 to 45°.
It is preferable to culture aerobically for 10 to 96 hours in the range of C.

(R,5)-8-halogeno-1,2-propanediol is treated with microorganisms to produce (R)-8-halogeno-1,2-
Propanediol can be obtained by adding a substrate to the culture solution cultured as described above or to a suspension of bacterial cells obtained from this culture solution by centrifugation, or by adding a substrate to the medium and culturing and reacting simultaneously. There are two methods: a method in which a substrate is added to a suspension of microorganisms immobilized by a conventional method in an appropriate buffer solution, and the like. The reaction temperature is 15~
The pH of the reaction at 50°C is preferably in the range 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 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, enzyme amount, and other reaction conditions, but it is preferable to select conditions so that the reaction is completed in 24 to 120 hours. To stop the reaction, the remaining substrate is analyzed by gas chromatography, etc., and the remaining substrate is about 50% of the added substrate.
From the viewpoint of yield, it is preferable to stop when the temperature reaches around 10%.

(R)-3-halogeno-1,2 thus obtained
-Propanediol can be collected from the reaction solution using a general method for collecting (R,S)-3-halogeno-1,2-propanediol. 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 over anhydrous sodium sulfate or the like, the solvent is removed under reduced pressure to obtain a syrup of (R)-8-halogeno-1,2-propanediol. Further, it may be further purified by distillation.

(Example) The following two examples will specifically explain the present invention.
1-37 Glucose 4%, (NH4)2HPO4, 1.8%,
KH2PO40.7%, MgSO4・7H20800p
p.m.

Zn5Os4H2060ppmX FeSO44H20
90ppmS CuSO4・5H205ppm, MnS
O4,4H2010ppm, NaC1100ppm,
A medium consisting of 0.3% yeast extract (pH 7.0) was prepared with deionized water, and 500 liters each was dispensed into 2β slope flasks and sterilized at 120°C for 20 minutes.

Each of the bacterial strains shown in Table 1 was inoculated into the above medium, and cultured with shaking at 30°C for 48 hours to obtain a culture solution 1.51.

This culture solution was centrifuged to collect bacterial cells, and after washing with water, the bacterial cells were reduced to 0.8 M! (R,S)-3-chloro-1,2-
2g of propanediol was added and reacted at 30'C for 72 hours with shaking.

Sterilization of 500 reactions/reaction liquid was performed by centrifugation, and the supernatant was
It was concentrated to 0 g/ml and extracted three times with 150 g/ml of ethyl acetate (total of 4505 g/ml). The extract was dehydrated over 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.

In addition, after each syrup was tosylated by a conventional method, an optical isomer separation column [Chiralcel O1C column (0,4
6c! llX25aI&)] (manufactured by JASCO Corporation) using a solvent of hexane-isopropyl alcohol (95:5), flow rate 1.0 tttl/min, wavelength 235 nm.
HPLC analysis under the following conditions (retention time 35 minutes for 8 bodies, 38 minutes for 2 bodies)
．． 5 minutes), it was confirmed that each of the samples contained the corresponding (R) isomer.

Table 1 Table 1 continued Table 1 continued However, the yield is (R,S)-3-chloro-1,
Determined from 2-propanediol. In addition, only in Example 37, a medium (pH 7.0) consisting of 2% glucose, 2% glycerin, 2% Corn Steepley's strength, 0.3% yeast extract, and 1% calcium carbonate was used, but in other cases The conditions were the same as in Examples 1-36.

Examples 38 to 74 Using the bacterial strains shown in Table 1, changing the substrate to (R,S)-3-bromo-1,2-propanediol, and otherwise performing the same operations as in Examples 1 to 37, Table 2 The results shown are obtained.

Table 2 Table 2 continued Table 2 continued However, the yield is the added (R,S)-8-bromo-1,
Determined from 2-propanediol. Further, for the culture in Example 74, the same medium as in Example 37 was used.

Patent applicant Kanebuchi Chemical Industry Co., Ltd. Agent Patent Attorney Makoto Asano - Procedural Amendment IO month 31, 1986

Claims (4)

[Claims] (1) Endomyces genus, Sporidiobolus genus, Geotrichum genus, Hansenula (H
an-senula), Rhodaromyces (Lodder)
Kluyveromyces genus, Kluyver
omyces), Nadsonia, Pichia, Saccharomyces
charomyces), Satucharomycopsis (Sa
ccharomycopsis), Rhodosporidium (Rhodosporidium), Rhodotorula (
Genus Rhodotorula, Wic
kerhamia), Ca-ndid
a) Genus Stepha-noas
cus), Torulopsis, Pachysolen, Schizosaccharomyces
Genus, Sporobolomyces
) genus, Brettanomyces
Genus, Escherichia, Aeromonas, Ac-idiphilium, Micro-coccus, Bacillus
Enterobacter spp.
ter) genus, Nocardia genus, Protaminobacter
Genus, Pseudomonas, Ar-throbacter, Hafnia, Klebsiella
iella) genus, Rhodococcus
s) genus, Gluconobacter
r) Microorganisms belonging to the genus (R,S)-3-halogeno-1
, 2-propanediol and the remaining (R)-
Optically active (R)-3-halogeno-1,2 characterized by collecting 3-halogeno-1,2-propanediol
-Production method of propanediol. (2) The microorganism is Endomyces magnusii (En-do).
myces magnusii), Sporidiobolus joh
nsonii), Geotrichum candidum (Ge
otrich-um candidum), Hansenula anomala
, Rhodaromyces elongysporus (Lodderom)
yces elong-isporus), Kluyveromyces fragilis (Kluyveromyces f.
ragilis), Nadsonia elongata (Nad
sonia elongata), Pichia burtonii, Saccharomyces cerevisiae
isiae), Satucharomycopsis lipolyteica (S
acchromycopsis lipolytica
), Rhodosporidium toruroides (Rhodo-
sporidium toruloides), Rhodotorula rubra
, Wickerhamia fluorescens
mia fluorescens), Candida humicola, Candida utilis
s), Stepha Noascus Schifferly
noascusciferrii), Torulopsis grope
ngiesseri), Pasisoren tannophyllus (
Pachysolentannophilus), Schizosaccharomyces pombe (Schizosacchar
omyces pombe), Sporobomyces salmonicolor (Sporobomyces sal)
monicolor), Brettanomyces custersianus (Brettanomyces custersi)
anus), Es-cheric
hia coli), Aeromonas hydrophila (
Aeromonas hydrophila), Acidiphilium cryptum (Acidiphilium-iu)
M cryptum), Micrococcus luteus (M
icrococcus luteus), Bacillus cereus, Enterobacter aerogenes
aerog-enes), Nocardia globulella (
Noc-ardia globerula), Protaminobacter alboflavus (Protaminoba
cter albo-flavus), Pse-udomonas fragi,
Pseudomonas crucibiae
cruciviae), Pseudomonas chlororaphuis (Pseudomonas chlororapis)
his), Arthrobacter simplex (Art
hrobacter simplex), Hafnia alvei, Klebsiella
Klebsiellapneumonia
as), Rhodococcus erythropolis (Rhodoc
occus erythropolis), Gluconobacter suboxidans (Glucon-obacter)
er suboxydans). (3) Claim 1 or 2, wherein the (R,S)-3-halogeno-1,2-propanediol is (R,S)-3-chloro-1,2-propanediol. manufacturing method. (4) Claim 1 or 2, wherein the (R,S)-3-halogeno-1,2-propanediol is (R,S)-3-bromo-1,2-propanediol. manufacturing method.