JPS61271992A - Production of optically active diol - Google Patents

Abstract

PURPOSE:To make it possible to produce a diol even from an electron attractive substituent group and in the presence of a saccharide, by using a specific microoragnism of the genus Saccharomyces. CONSTITUTION:A microorganism belonging to the genus Saccharomyces, and having the ability to produce a 1-arylpropane-1,2-diol, e.g. Saccharomyces cerevisiae IFO 0203 or Saccharomyces cerevisiae IFO 0233, or a baker's yeast such as Oriental Dry Yeast (manufactured by Oriental Yeast Co., Ltd.) or Super Camellia (manufactured by Nisshin Flour Milling Co., Ltd.), is cultivated in a culture medium containing benzaldehyde or a substituted benzaldehyde expressed by formula I (X is Cl, F or methyl group at the o-position, F group at the m-position or nitro, Cl, F, methyl, methoxy group, etc. at the p-position) and a saccharide to produce a (1S,2R)-arylpropane-1,2-diol expressed by formula II, which is then collected.

Description

Detailed Description of the Invention (Industrial Application Field) The present invention relates to a method for producing optically active (+13,2R) -1-arylpropane-1,2-diol, and more specifically, to a method for producing an optically active (+13,2R) -1-arylpropane-1,2-diol, and more specifically, to .2R)-1
- After culturing bacteria capable of producing arylpropane-1,2-diol in water containing benzaldehyde or substituted benzaldehyde and sugar,
R) General formula for collecting -1-arylpropane-1,2-diol (in the formula, X is a substituent bonded to the rose position, meta position, or ortho position, and is O-chloro or O-fluoro.

This is a method for producing diols represented by 0-methyl, m-fluoro, p-nitros, p-chloro, p-fluoro, p-methyl, p-methquin, etc.

The diol obtained in the present invention is a raw material for pharmaceuticals such as optically active tatanosamine and ristosamine [for example, Journa
l of Organi c C! hami 5try
, Vol. 48, p. 909 (1983)], raw materials for bioactive compounds such as ephedrine (Journal of
AmericanC! chemical 5ociet
y, Vol. 79, pp. 197, 201 (1957)
] It is a useful compound that can be obtained.

(Prior Art) Conventionally, methods for producing diols are broadly classified into organic chemical methods and biochemical methods using oxygen and microorganisms. Generally known organic chemical methods include a method in which an olefin is converted into an epoxide using a peracid and then ring-opened, and a method in which the olefin is oxidized with potassium permanganate.

(Problems to be Solved by the Invention) However, these methods have drawbacks in that the oxidizing agent is explosive and dangerous, and it is difficult to produce optically active substances. As a biochemical method using microorganisms and dust particles, Saccharomyces cereviinae (8acchar
A method is known for producing (1) from benzaldehyde by opening a strain of P. omyce6 Cereviciae. (Chemistry and Industry
, p. 983 (1977)]. However, if this method is used, a compound in which X in (1) is hydrogen and p-methoxy will produce a diol, but if an electron-withdrawing group such as a chloro group or a nitro group is combined at the p-position, It was thought that it was impossible to produce diols.

(The rounded hand M that solves the problem!) The present inventors searched for a strain that can produce diol from benzaldehyde that has an electron-withdrawing substituent, and found that
In the presence of sugars, using a bacterial strain that belongs to the Buttucharomyces genus,
1! The present invention was completed by discovering that diols can be produced in good yield from benzaldehydes that are child-donating or absorbing, or from various benzaldehydes having substituents. Note that the operation according to the present invention is extremely simple and easy to set reaction conditions as compared to conventional methods, and has excellent features in that diol Ll) can be obtained in good yield.

The substituted benzaldehyde used in the present invention has the general formula (II) (wherein, X is O-chloro, 0-fluoro, o-methyl, m-fluoro, p-nitro% p-chloro, p-fluoro, p-methyl , p-methoxy, etc.), specifically, 0-chlorobenzaldehyde, 0-fluorobenzaldehyde, 0-methylbenzaldehyde, m-fluorobenzaldehyde, p-chlorobenzaldehyde, p- Fluorobenzaldehyde, p-nitrobenzaldehyde.

Examples include substituted benzaldehydes such as p-methylbenzaldehyde and p-methoxybenzaldehyde, but are not limited thereto.

Benzaldehyde substituted with pressure heat can also be used.

The bacteria used in the present invention belong to the genus Bucharomyces.1 A typical example is Saccharomyces Cerevisiae.
iae) IFO0205, Satucharomyces φ serequisiae y.

Commercially available baker's yeast such as 0233, Oriental Dry Yeast (Oriental Yeast), and Super Camellia (Nissin Seifun) can be used.

The reaction is carried out by stirring a mixture of these yeast extracts, water, and sugars such as glucose and sucrose at a temperature of 20 to 40°C. 1. When fermentation begins, the substrate benzaldehyde or substituted benzaldehyde is added to the mixture or as an ethanol solution. This is done by adding and stirring for several hours to three days.

Further, when 0.3 to 1.2 t of acetaldehyde is added to the reaction solution per 1 t of substrate, reduction to benzyl alcohol can be prevented and the yield can be improved. pH of medium
A score of 5 to 10 is appropriate.

7 to 9 are more preferred. The concentration of the substrate to be used is not particularly limited, but is generally preferably about 0.1 to 5%. Diols are isolated from the culture solution after removing bacterial cells by centrifugation or suction filtration.

Alternatively, for example, use the culture solution without removing the bacterial cells.

Extraction with an organic solvent such as ethyl acetate or ethyl ether, followed by nine-column chromatography using silica gel, alumina, etc. as a packing material, centrifugal chromatography using silica gel, alumina, etc. as an adsorbent, distillation, recrystallization, or other conventional purification methods. Purify using.

Hereinafter, the present invention will be explained in more detail with reference to Examples.
The present invention is not limited to these.

Example 1 Phosphorus salt buffer 50m pH 9, dry yeast (Oriental Tri Yeast) 12.5F, glucose 10
The mixture was weighed into a ft-5001 L eggplant flask and stirred at room temperature for 10 minutes. In this, benzaldehyde 6101
IP (5.75 mmol) in ethanol solution (1s+
l) and acetaldehyde 480 q (11 mmol
) and stir using a mug mixer at room temperature.

After 1 hour, add dry yeast (Oriental dry yeast)
6.3 t and glucose 52 were added and further heated at room temperature for 3.
Stirred for 5 hours. Then, the reaction solution was poured into a beaker containing 35 tons of Celite, filtered and filtered, and extracted three times with 100 mg of ethyl acetate. The organic layers were combined and dried over anhydrous sodium sulfate, and the solvent was distilled off under reduced pressure. After death, silica gel column chromatography (eluent: hexane-ethyl acetate = 4:r) was performed to extract (IR, 2B)-1-phenylpropane. 385 Misaki was obtained by using an optically active form of -1,2-diol as an oil. The yield was 44%.

Infrared absorption (Neat) air' 5400, 3080.2900.1650.1500.
1450°+380.1200. +130.1080
．． 1040. 990° nuclear magnetic resonance absorption (cc14) δppm = 1.07 (d, 3H)2, 77 (
broad a, 2H) 4, Oo (axquart
IH) 4.63 (d, IH) 7, 16-7.5 (m, 5H) Mass spectrometry spectrum (m/z, rel 1ntens
ity)+35(99), 108(100), 79
(88), 45 (16), 27 (specific optical rotation [α) D-38,9° (C!=2.05. C!HO
1,) Example 2 Dry yeast 12.5 f, glucose IO? , tap water 50
d was weighed into an eggplant flask and stirred at room temperature for 10 minutes. Add 0.605 f of p-chlorobenzaldehyde to this.
(3.24 mmol) of ethanol solution was added thereto, and the mixture was stirred at tg temperature for 1 hour. Here, 6.5 tons of dry yeast
and glucose 5f were added, and the mixture was further stirred at room temperature for 3.5 hours. Next, the reaction solution was poured into a beaker containing 35 tons of Celite, and extracted with ethyl acetate in the same manner as in Example 1.
After drying over anhydrous sodium sulfate and distilling off the solvent, (IR, 28) -1-(
0.220 f of an optically active form of p-chlorophenyl)-globan-1,2-diol was obtained as an oil. The yield was 27X.

Infrared absorption (Neat) an'' 3400, 3000.2950.1910.1720.
1600°1500, +410.1200. 930
．． 850. 800°740, 720 Nuclear magnetic resonance absorption (cc14) δppm = 0.95 (d, 3H) 3, 15 (broad 8.2H) 3.85 (d X quart, IH) 4.59 ((
1, In) 7.08-7.43 (m, 4H) Mass spectrometry spectrum (+!!/2. rel 1nte
nsity) 186(2), 169(2), 1
41 (100), +07 (1).

77(40), 45(9), 28(17) The specific optical rotation [α] is also j −37,6° (C!=2.00.
cHcl, ) Box Example 3 Dry yeast + 2.5 f, Glue: ff -su 10 p, and tap water 50 d were weighed into an eggplant flask and stirred at room temperature for 10 minutes. To this, 0-chlorobenzaldehyde 0.607
? (4.32 mmol) of ethanol solution was added thereto, and the mixture was stirred at room temperature for 1 hour. Here dry yeast 6
．． 32 and Cucrose 5f were added, and the mixture was further stirred at room temperature for 3.5 hours. Next, the reaction solution was poured into a beaker containing 3 tons of Celite, extracted with ethyl acetate in the same manner as in Example 1, dried over anhydrous sodium sulfate, distilled off the solvent, and subjected to silica gel column chromatography (eluent :hexane-ethyl acetate = 4:+) (IFI, 2B) -1
An optically active form of -(0-chlorophenyl)-propane-1,2-diol was obtained as an oil with a weight of 0.256 F. The yield was 529.

Infrared absorption (Neat) cIn4 3400, 3000.2950. +710.1600
．． 1570°3440, 1200. 930. 850
．． 760. 710 Nuclear magnetic resonance absorption (ccx,) δppm = 0.90 (d, 3H) 3.70 (broad S, 2H) 4.16 ((I X quart, IH) 5.13 (
a, 11) 7.01-7.0 (m, 3H) 7.43-7.75 (m, IH) Mass spectrometry spectrum (le'z rel 1ntensi
ty) 186(1), 169(2), 152(2
), 141 (59), 105 (44 near 7 (10
0), 45 (47), 29 (22), 14 (60
) Specific optical rotation (α)''; -31,8° (C!=1.390H
CIg) Example 4 Dry yeast +2.5f, glucose 102, tap water 50d
(weighed into an eggplant flask) and stirred at room temperature for 10 minutes.

To this, 0.6049 p-fluorobenzaldehyde (
4.87 mmol) of ethanol solution (1 d) was added thereto, and the mixture was stirred as it was at room temperature for 1 hour. Here, 6.3v of dry yeast and 5f of glucose were added, and the mixture was stirred at room temperature for 5.5 hours. Next, Celite 35? The reaction solution was poured into a beaker containing 100% chloride, extracted with ethyl acetate in the same manner as in Example 1, dried over anhydrous sodium sulfate, the solvent was distilled off, and then subjected to silica gel column chromatography (eluent: hexane-ethyl acetate = 4 :1) to obtain 0.214 as an optically active form of (IR, 2B) -1-(p-fluorophenyl)-propane-1,2-diol as an oil. The yield is 2
It was 6%.

:.

Infrared absorption (Neat) arr' 3400.3000,2900. +900.1700.
1605°1510, +450. 1220. 9
30. 850. 820° nuclear magnetic resonance absorption (ccl,
) δppm = 0.90 (d, 3H)5, 80 (
dXquart, IH) 4.23 (broad S
, 2H) Now, 53 (d, IH) 6.73-7.50 (m, 4H) Mass spectrometry spectrum (n1/z rel 1ntens
ity) 170 (υ, 9), +53 (14),
+25 (100).

97 (51), 77 (15), 45 (+3), 2
8 (22) Specific optical rotation (a)"a' -ss, 9° (a = 2.Of c
aclg) Example 5 Dry yeast 12.5r, glucose! Of, tap water 501
The mixture was weighed in an IJt-Eggplant flask and stirred at room temperature for 10 minutes. To this, m-fluorobenzaldehyde 0.605
9 (4.88 mmol) in ethanol was added thereto, and the mixture was stirred at room temperature for 1 hour. Here dry yeast 6.
32 and glucose 52 were added, and the mixture was further stirred at room temperature for 3.5 hours. Then, the reaction solution was poured into a beaker containing 35 tons of Celite, extracted with ethyl acetate in the same manner as in the previous example, dried over anhydrous sodium sulfate, and the solvent was distilled off.
(IR, 2B) -l-m by silica gel column chromatography (eluent: hexane-ethyl acetate = +:1)
0.263 r of an optically active form of -fluorophenyl-propane-12-diol was obtained as an oil. The yield was 31%.

Infrared absorption (Neat) equipment-1 5400, 5000, 2950, 16+5.1590.
+490°1450, 1250. +130.107
0. +040.1000° nuclear magnetic resonance absorption (cc 1
4) δppm = 0.93 (d, 3H) 349 (bro
ad El, 2H) 3,85 (dXquart, IH) 4.65 (d, IH) 6.63-7.23 (m, 4H) Mass spectrometry spectrum (m//zrθ11ntensit
y) 170(0,3), 152(2), 126
(100), 105 (2s.

97 (81), 77 (16), 45 (20),
28 (N) Specific optical rotation (a piece' -2a, s° (0 = 1.97 (HC
I,) Example 6 Dry yeast 12.5f, Glue: l-su IOS', tap water 5
0d was weighed into an eggplant flask and stirred at N temperature for 10 minutes. This GCO-Fluoropenzartecht 0.602
1 d of an ethanol solution of f (4.85 mmol) was added, and the mixture was stirred at room temperature for 1 hour. Here, 6.32 ml of dry yeast and 52 ml of glucose were added, and the mixture was further stirred at room temperature for 3.5 hours. Next, the reaction solution was poured into a beaker containing Celite 35f, extracted with ethyl acetate in the same manner as in Example 1, dried over anhydrous sodium sulfate, distilled off the solvent, and subjected to silica gel column chromatography (eluent:
to Φsan-ethyl acetate = 4:I) (IR, 2B) -
The optically active form of 1-(0-fluorophenyl)-70by-1,2-diol as an oil is 0.245? Obtained.

The yield was 50%.

Infrared absorption (Neat) an-' 3400, 3000.2950.1620.1585.
1490°1450, 1220.1+80.1130.
990. 930° nuclear magnetic resonance absorption (CIC1*) δppm = 0, 90 (d, 3H) 193 (
m, 2H+1H) 4.95 (d, IH) 6.80-7.63 (m, 4H) Mass spectrometry spectrum (aI/z rel 1ntens
ity) 170(0,5), 152(2), 12
5 (100), 105 (52).

97 (59), 77 (35), 45 (22), 2
g(70) Specific optical rotation [α) D −24,s° ((! = 2.440H
C'1. ) Example 7 12.5 f of dry yeast, 50 g of glucose, and 50 d of tap water were weighed into an eggplant flask and stirred at room temperature for 10 minutes. To this, p-methylbenzaldehyde 0.606
1 d of an ethanol solution of f (5.05 mmol) was added, and the mixture was stirred at room temperature for 1 hour. Here dry yeast 6.
32 and glucose 5f were added, and the mixture was further stirred at room temperature for 3.5 hours. Next, Celite 35? The reaction solution was poured into a beaker containing , extracted with ethyl acetate in the same manner as in Example 1, dried over anhydrous sodium sulfate, and the solvent was distilled off.
(IR, 2B) - by silica gel column chromatography (eluent: hexane-ethyl acetate = 4: +)
0.255 f of an optically active form of 1-(p-methylphenyl)-propane-1,2-diol was obtained as an oil.

The yield was 28%.

Infrared absorption (N8at) cm-'3400 3
000 2950 1920 +720 1
7201710 +620 1520 145
0 1380 12001s5o, jooo,
930. 860. 84o, goo.

Nuclear magnetic resonance absorption (cc14) δppm = 0.96 (d, 5H) 2.33 (S, 3H) 2.83 (broad S, 2H) 3.80 (dXquart, IH) 4.53 (d,
IH) 6.87-7.35 (m, 4H) Mass spectrometry spectrum (m/z rel tntensl
ty) 166(0,5), +49(Isu, 121(
100), 93(48).

? ? (28), 43(8), 28(7) Specific rotation [α]! Also' -42,2° (0= 2.10
cHclm) Example 8 12.5 f of dry yeast, lof of GuA course, and 50 d of tap water were weighed into an eggplant flask and stirred at room temperature for 10 minutes. Add O-methylbenzaldehyde 0,6+3
1 d of an ethanol solution of f (5,II mmol) was added, and the mixture was stirred at room temperature for 1 hour. Here dry yeast 6
．． 32 and glucose were added, and the mixture was further stirred at room temperature for 3.5 hours. Next, the reaction solution was poured into a beaker containing Celite 55V, extracted with ethyl acetate in the same manner as in Example 1, dried over anhydrous sodium sulfate, distilled off one solvent, and then subjected to silica gel column chromatography (eluent: hexane). Ethyl monoacetate = 4: J) (IR, 2S)
0.127 f of an optically active form of -J-(0-methylphenyl)-propane-1,2-diol was obtained as an oil. The yield was 7%.

Infrared absorption (Neat) country-1 5400, 3000, 2950, 1700, 1450,
1380°50. +070. 990. 950.
850. 750 Nuclear Magnetic Resonance Absorption (0014) a ppm = +, 05(d, 3H)2jO(S
, 3H) 2.82 (broad 8, 2H) 5.83 (dXquart, IH) 4.83 (d, IH) 6.83 ~ 7.23 (m, 5H) 7.3G ~ 7.50 (m, IH) Mass spectrometry spectrum (Le/2 red intθnθ1
ty) 182(5), 164(3), +37(
100), +21 (+9).

+09 (+9), 94 (eyes), 77 (10), 2
B(39) Specific optical rotation [α)”; −55,9° (0= 1,020
HO'ls) Example 9 Dry yeast 12.5? , glucose lof, tap water 50
d was weighed into an eggplant flask and stirred at room temperature for 10 minutes. To this, p-methoxypenzaltehyde 0.608 F
(4.47 mmol) of ethanol solution 1- was added,
The mixture was stirred for 1 hour at 11°C. Here dry yeast 6.
3f and glucose 5f were added, and the mixture was further stirred at room temperature for 345 hours. Next, the reaction solution was poured into a beaker containing Celite 35V, and extracted with ethyl acetate in the same manner as in Example 1, dried over anhydrous sodium sulfate, and the solvent was distilled off.
(IR, 2B) -1-(
0.183 was obtained as a crystal of an optically active form of p-methoxyphenyl)-propane-1,2-diol. The yield was 23%, melting point 94-95°C.

Infrared absorption (l θat) m-' 3450, 3350.2970.1720.1610.
1580°+510.1450.1410.1240.
1080.1020°990, 860. 840.
800. 670. 580° nuclear magnetic resonance absorption (cc1
4) δppm = 1.08((1,3)1)2.0(br
oad S, 2H) 3.73-4.10 (m, 3H) 4.90 (d, IH) 6.87 (d, 2) i) 7.27 (d, 2H) Mass spectrometry spectrum (m/z rel 1nten++
1ty) 1i(0,2), +49(2), 122(
100), 107(54).

93(71), 77(31), 45(8), 28
(38) Specific rotation [α] -3-35.9° (a: 1,02 cH
clg) Example 10 Dry yeast 12.5F, glucose IOf, tap water 50d
The mixture was weighed out into an eggplant flask and stirred at room temperature for 10 minutes.

Add p-nitropenzaltehyde 0.601 to this? (
3.98 mmol) of ethanol solution 1- was added thereto, and the mixture was stirred with Na for 1 hour. Here, 6.52 ml of dry yeast and 52 ml of glucose were added, and the mixture was further stirred at room temperature for 3.5 hours. Then, the reaction solution was poured into a beaker containing 35 tons of Celite, extracted with ethyl acetate in the same manner as in Example 1, dried over anhydrous sodium sulfate, distilled off the solvent, and subjected to silica gel column chromatography (eluent: hexane-hexane). The optically active form of (IR, 2B) -1-(p-nitrophenyl)-propane-1,2-diol was prepared as an oil using ethyl acetate=+:+). I got +099. The yield is 14
%Met.

Infrared absorption (Neat) cm-' 300. 3000. 2900. +940
．． 1605. 1520°13'+0.1200
．． 930. 11140. 810. 750° Nuclear Magnetic Resonance Absorption (OC14) δppm = 1.05 (d, 3H) 2.86 (1,1road S, 2H) 4.10 (dX
quart, IH) 7.50 (d, 2H) 8° + 6 (d, 2H) Mass spectrometry spectrum (m/z rel 1nto-sty) +98 (0,6), +81 (su, +64 (
0,8), +53 (100).

+56 (83), +06 (52), 77 (21),
45 (26).

Specific optical rotation (α) -107,2° (C= 1.+54 c
Hcx,) Patent Applicant Central Glass Co., Ltd. Procedural Amendment Homare (Voluntary) January 1985 Special FF Agency Director General Ubuyaki Michibe 1, ◆Indication of 1985 Special FF Issue No. 3159 2 Name of the invention Process for producing optically active diols 3 Relationship with the case of the person making the amendment Patent applicant address 5253% Oki-Ube, Ube-shi, Yamaguchi Prefecture Name (2)
20) San Ito, Representative of Centcil Glass Co., Ltd.
5. Number of inventions increased by amendment None 6.
Claims column of the specification subject to amendment and details of the invention Ii5! Column 7, Contents of the Amendment "Written Amendment" and Feature a' are amended to the extent that the scope of the claims is as shown in the appendix.

A bacterium belonging to the genus Saccharomyces and capable of producing l-aryl7opan-1,2-diol was cultured with stirring in the presence of benzaldehyde or substituted benzaldehyde and sugars, and from the culture (IR , 28-1-arylgloban-1,2-diol (IR, 28
) -Method for producing 1-arylpropane-1,2-diol, λ The Detailed Description of the Invention column is edited as follows.

(Li Specifications page 1, line 15, and line 17, 4 layers 2
Correct the 1-(1S, 2R month of Teiju 2 line to r(IR, 2s month).

(2) Amend the ``general formula for sampling'' on page 2, line 5 of the same document to read ``take 1 sample''.

(4) On page 2, line 5, [(in the formula, X is] the ku place)
Correct it with 1 (in the formula or hydrogen or) (position, j).

(5) 1p-Metkin, etc. in the 8th to 9th lines of the 2nd Sada)
" is corrected to "p-methoxy, etc.)".

(6) 1°Oxygen in the 3rd row of the same prefecture and the 1st O to 11th lines is corrected to ``# element J.

(7) Correct "X is 0-chloro," in the first line of No. 5, to "rx is hydrogen or O-chloro."

(8) +-(m/2.jt-r on page 10, line 7)
(Correct as m/Z and J.

Claims (1)

[Claims] A bacterium belonging to the genus Saccharomyces and capable of producing 1-arylpropane-1,2-diol is cultured with stirring in the presence of benzaldehyde or substituted benzaldehyde and sugars, and from the culture (1S
,2R)-1-Arylpropane-1,2-diol is collected. Optical activity (1S,2R)-1
- Method for producing arylpropane-1,2-diol.