JP3067817B2 - Method for producing optically active (-)-2-halo-1- (substituted phenyl) ethanol - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

The present invention relates to (-)-2-halo-1-
More specifically, the method for producing (substituted phenyl) ethanol is described in detail by contacting a microorganism with 2-halo-1- (substituted phenyl) ethanone to efficiently produce (-)-2-halo-1- (substituted phenyl) ethanol. It relates to a method of manufacturing. This compound or (-)-substituted styrene oxide obtained by ring-closing the compound under alkaline conditions is useful as a raw material for synthesizing pharmaceuticals, agricultural chemicals and the like that require optical activity.

[0002]

2. Description of the Related Art The present inventors have not found any patents, reports or the like showing the method for producing optically active (-)-2-halo-1- (substituted phenyl) ethanol.

[0003]

SUMMARY OF THE INVENTION The inventors of the present invention have studied to develop optically active (-)-2-halo-1- (substituted phenyl) ethanol, and as a result, have found that 2-halo-1- (substituted Stereospecific asymmetric reduction of phenyl) ethanone to give (-)
The present inventors have found that there is a microorganism capable of converting -2-halo-1- (substituted phenyl) ethanol, and completed the present invention.

[0004]

The first aspect of the present invention is as follows.
General formula [1]

[0005]

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(Wherein, X represents a chlorine atom or a bromine atom, and the substituents R ₁ , R ₂ , and R ₃ represent a hydrogen atom, a chlorine atom, a fluorine atom, a methyl group, or a methoxy group. 2-halo-1- (substituted phenyl) ethanone represented by the general formula [2]:

[0007]

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(Wherein X and the substituents R ₁ , R ₂ , R ₃ are the same as in the general formula [1], and * represents an asymmetric carbon atom). Acibia, Brettanomyces, Pichia, Rhodosporidium, Trigonopsis, Cryptococcus alvidas, Cryptococcus terreus, Candida intermedia, Candida with the ability to asymmetrically reduce to (substituted phenyl) ethanol Selected from the group of microorganisms belonging to Crusay, Candida magnoliae, Candida pinus, Candida cytoana, Candida tropicalis, Rhodetorla graminis, Rhodetorla minuta, Rhodetorla rubra, Saccharomyces cerevisiae It is represented by the general formula [2] produced by contacting with microorganisms ( ) -2-halo-1- (and collecting the substituted phenyl) ethanol (-) - 2
-Halo-1- (substituted phenyl) ethanol.

The 2-halo-1- (substituted phenyl) ethanone used in the present invention is asymmetrically reduced to give (-)-2-halo-1-
Microorganisms that convert to (substituted phenyl) ethanol can be found by the methods described below. For example,
40 g glucose, 3 g yeast extract (NH ₄ ) ₂ HPO ₄ 1
3 g, KH ₂ PO ₄ 7 g, MgSO ₄ .7H ₂ O 0.8 g, ZnSO ₄ .7H _{2
O 60mg, FeSO 4 · 7H 2} O 90mg, CuSO 4 · 5H 2 O 5mg, Mn
_{SO 4 · 4H 2 O 10mg,} NaCl0.1g ( per liter)
50 ml of the A medium having the above composition was placed in a 500 ml Sakaguchi flask, sterilized, and then inoculated with microorganisms, followed by shaking culture at 30 ° C. for 2 days. Thereafter, the cells were collected by centrifugation and 2-bromo-
0.1 M phosphate buffer (pH 7.0) containing 0.5% of 1- (3'-chlorophenyl) ethanone and 3% of glucose 2
Suspend in 5 ml and shake at 30 ° C. for 2-3 days in a 500 ml Sakaguchi flask. Thereafter, an equal amount of ethyl acetate was added to perform extraction, and the resulting (-)-2-bromo-1- (3'-chlorophenyl) ethanol was subjected to gas chromatography (column: silicon OV-7, φ0.3 × 200 cm, column temperature). The analysis is performed at 190 ° C. under a N ₂ gas pressure of 1.2 kg / cm ² ). The optical purity of (-)-2-bromo-1- (3'-chlorophenyl) ethanol was determined by distilling and extracting the extracted oil.
High performance liquid chromatography (Column: Chiralcel-OJ, manufactured by JASCO Corporation, elution solvent hexane-isopropanol (50: 1), flow rate 1.0 ml / min, detection 220 n
According to m), the (-) form is 44.8 minutes and the (+) form is 54.9 minutes.
Separation is achieved with a retention time of minutes and the optical purity can be determined.

The microorganism which can be used in the present invention includes 2-
Any microorganism capable of asymmetrically reducing halo-1- (substituted phenyl) ethanone to (-)-2-halo-1- (substituted phenyl) ethanol can be used. For example, Ashbya gossypii
IFO 0560, Brettanomyces Castellicyanus (Brett
anomyces custersianus) IFO 1585, Candida intermedia IFO 0761, Candida krusei IFO 0011, Candida magnoliae IFO 070
5, Candida pinus (Candida pinus) IFO 0741,
Candida saitoana IFO 076
8.Candida tropicalis
IFO 1403, Cryptococ Alvidas
cus albidus) IFO 0378, Cryptococcus terreus
(Cryptococcus terreus) IFO 0727, Pichia farinosa IFO 0574, Pichia membranaefaciens IFO 0460,
Rhodosporidium
toruloides) IFO 0871, Road Torla Graminis (Rho
dotorula graminis) IFO 0190, Lord Torla Minuta
(Rhodotorula minuta) IFO 0387, Rhodetorula rubra IFO 0383, Saccharomyces
Saccharomyces cerevisiae IFO 0614, Trigonopsis variabilis
IFO 0671 or the like can be used.

In culturing these microorganisms, usually any nutrient source that these microorganisms can utilize can be used.
For example, glucose, carbohydrates such as sucrose, ethanol, alcohols such as glycerol; hydrocarbons such as paraffin, organic acids such as acetic acid, propionic acid; carbon sources such as soybean oil or mixtures thereof, yeast extract, peptone, meat extract, Nitrogen-containing inorganic organic nutrients such as corn steep liquor, ammonium sulfate, and ammonia; phosphates, magnesium, iron,
A normal medium appropriately containing an inorganic nutrient such as manganese and potassium, and vitamins such as biotin and thiamine is used. As a culture method, the nutrient medium is cultured aerobically at a pH of 4.0 to 9.5 and at a temperature of 20 to 40 ° C for 1 to 5 days.

[0012] As a method for reduction, a method using the culture solution as it is, centrifugation or the like to separate the cells, resuspend the cells in a phosphate buffer or water, etc., and prepare 2-halo-1
There is a method of adding and reacting-(substituted phenyl) ethanone. During this reaction, a carbon source such as glucose or sucrose may be added as an energy source. The cells may be living cells, or may be treated with acetone, freeze-dried, or the like. In addition, these cells can be immobilized on a carrier and used. 2-halo-
The 1- (substituted phenyl) ethanone may be added as it is, or may be dissolved in an organic solvent so as not to affect the reaction and added all at once from the beginning of the reaction or added in portions. The reaction is carried out at a temperature of 10 to 60 ° C. and a pH of 3 to 120 at a pH of 5 to 9.
Perform under stirring for hours.

(-)-2-halo- formed by the reaction
1- (Substituted phenyl) ethanol can be collected from the reaction solution directly or after isolation of cells, extraction with a solvent such as ethyl acetate or dichloromethane, dehydration, and purification by distillation or silica gel chromatography or the like. −)
-2-Halo-1- (substituted phenyl) ethanol is readily obtained. Further, the optical purity is determined by the column, Chiral cel-OJ
, Elution solvent hexane / isopropanol (30-50
/ 1) can be measured in the same manner as described above by high performance liquid chromatography.

In the formula [2] obtained as described above, (-)
-2-Halo-1- (substituted phenyl) ethanol is converted to NaOH
And the like, and coexist in an equimolar or more amount, and the ring is easily closed by heating or standing at room temperature to obtain the (-)-of formula [3].
It can be converted to a substituted styrene oxide.

[0015]

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(The substituents R ₁ , R ₂ and R ₃ are the same as those in the general formulas [1] and [2], and * represents an asymmetric carbon atom.)

[0017]

The present invention will be described in more detail with reference to the following Examples, but it should not be construed that the invention is limited thereto. In the following description, “%” means “% by weight” unless otherwise specified.

Example 1 50 ml of the above-mentioned A medium was placed in a 500 ml Sakaguchi flask, sterilized, and inoculated with the microorganisms shown in Table 1. Then, shaking culture was performed aerobically at 30 ° C. for 2 days. The cells were collected from this culture by centrifugation, and 2-bromo-1-
(3′-chlorophenyl) ethanone 0.5% to 0.3%
0.1 M phosphate buffer (pH 7.0) containing glucose 25
suspended in a 500 ml Sakaguchi flask at 30 ° C.
The mixture was shaken for 48 hours. After the reaction, (-)-2-bromo-1- (3'-chlorophenyl) ethanol was extracted twice from the reaction solution with an equal volume of ethyl acetate, and the ethyl acetate layer was analyzed by gas chromatography to determine the reaction rate. Was. Next, ethyl acetate is dehydrated with anhydrous sodium sulfate, and the solvent is removed.
2-Bromo-1- (3'-chlorophenyl) ethanol was obtained. This was dissolved in methylene chloride and the optical purity was measured by high performance liquid chromatography. Table 1 shows the results.

[0019]

[Table 1]

The obtained (−)-2-bromo-1- (3 ′)
-Chlorophenyl) ethanol specific rotation [α] (2
0, D) -25.5 ° (c = 1.02 CH ₃ OH), (optical purity 100% by high performance liquid chromatography analysis e.
e.) H-NMR (90 MHz, CDCl ₃ ) δ ppm 2.88 (br. S, 1H), 3.3
5 to 3.90 (m, 4H), 4.90 (dd, J = 315, 8Hz, 1H) 6.98 to
7.51 (m, 4H).

Example 2 Using the microorganisms shown in Table 2, 2-bromo-1-substrate was used as a substrate.
A culture reaction and analysis were carried out in the same manner as in Example 1 except that 2-bromo-1- (2'-chlorophenyl) ethanone was used instead of (3'-chlorophenyl) ethanone.
(-)-2-Bromo-1- (2'-chlorophenyl) ethanol was obtained. Table 2 shows the results.

[0022]

[Table 2]

The obtained (-)-2-bromo-1- (2 ')
-Chlorophenyl) ethanol specific rotation [α] (2
0, D) -41.5 ° (C = 1.02, CH ₃ OH), (100% optical purity by high performance liquid chromatography analysis)
ee) H-NMR (90 MHz, CDCl ₃ ) δ ppm 2.78 to 3.06 (m, 1H),
3.39 (dd, J = 9, 10Hz, 1H), 3.73 (dd, J = 2.5, 10Hz, 1
H), 5.04 to 5.39 (m, 1H) and 6.68 to 7.68 (m, 4H).

Example 3 Using the microorganisms shown in Table 3, 2-bromo-1-substrate was used as a substrate.
A culture reaction and analysis were carried out in the same manner as in Example 1 except that 2-bromo-1- (4'-chlorophenyl) ethanone was used instead of (3'-chlorophenyl) ethanone.
(-)-2-Bromo-1- (4'-chlorophenyl) ethanol was obtained. Table 3 shows the results.

[0025]

[Table 3]

The obtained (-)-2-bromo-1- (4 ')
-Chlorophenyl) ethanol specific rotation [α] (2
0, D) -26.6 ° C (c = 1.10, CH ₃ OH), (100% optical purity by high performance liquid chromatography analysis)
ee) H-NMR (90MHz, CDCl ₃ ) δppm 2.77 (br, S, 1H),
3.18 to 3.70 (m, 2H), 4.82 (dd, J = 3.5, 7.5Hz, 1H),
6.82 to 7.44 (m, 4H).

Reference Example 1 Each of 10 g of (-)-2-halo-1- (chloro-substituted phenyl) ethanol was mixed with 5 ml of a 40% aqueous NaOH solution and 10 ml of methylene chloride and reacted at 50 ° C. for 6 hours. After cooling, methylene chloride (20 ml) was added, the methylene chloride layer was washed with saturated saline, dehydrated and filtered, and methylene chloride was removed under reduced pressure to obtain a crude epoxide oil. This was purified by distillation under reduced pressure to obtain each (-)-chloro-substituted styrene oxide shown in Table 4.

[0028]

[Table 4]

[0029]

According to the present invention, as shown in Examples, optically active (-)-2-halo-1- (substituted phenyl) ethanol can be produced efficiently.

──────────────────────────────────────────────────の Continued on the front page (51) Int.Cl. ⁷ Identification symbol FI (C12P 7/22 C12R 1:74) (C12P 7/22 C12R 1:74) (C12P 7/22 C12R 1: 865) (56 References European Patent Application Publication No. 198440 (EP, A1) Org. Chem. 45, no. 16 (1980) p. 3352-3355 (58) Field surveyed (Int. Cl. ⁷ , DB name) C12P 7/22 CA (STN) REGISTRY (STN)

Claims (3)

(57) [Claims] 1. A compound of the general formula [1] (In the formula, X represents a chlorine atom or a bromine atom, and the substituents R ₁ , R ₂ , and R ₃ represent a hydrogen atom, a chlorine atom, a fluorine atom,
It represents a methyl group or a methoxy group. However, the case where all three substituents are hydrogen atoms is excluded. The 2-halo-1- (substituted phenyl) ethanone represented by the general formula [2] (Wherein X and the substituents R ₁ , R ₂ , R ₃ are represented by the general formula [1]
And * indicates an asymmetric carbon atom).
Acibia, Bretanomyces, Pichia, Rhodespolidium, Trigonopsis, Cryptococcus albidus, Cryptococcus terreus, Candida, having the ability to asymmetrically reduce to 2-halo-1- (substituted phenyl) ethanol Intermedia, Candida Crusay, Candida Magnoliae, Candida Pinus, Candida Cytoana, Candida Tropicalis, Lord Torla Graminis,
(-)-2-halo-1- (substituted phenyl) represented by the general formula [2] produced by contacting a microorganism selected from the group of microorganisms belonging to Rhodotorula minuta, Rhodotorula rubra, and Saccharomyces cerevisiae. (-)-2-halo-1- characterized by collecting ethanol
A method for producing (substituted phenyl) ethanol. 2. The method according to claim 1, wherein the microorganism is Acibia gossipii, Bretanomyces castellicanus, Pichia farinosa,
2. The process according to claim 1, wherein the process is Pichia membrane aefaciens, Rhodes polydium toluroides, or Trigonopsis variabilis. 3. The method according to claim 1, wherein the microorganism is Ashbya gossypi.
gossypii) IFO 0560, Brettanomyces custersianus IFO 1585, Candida intermedia IFO 0
761, Candida krusei IFO 0
011-Candida magnolia
e) IFO 0705, Candida pinus I
FO 0741, Candida saitoan
a) IFO 0768, Candida tropicalis
ropicalis) IFO 1403, Cryptococcus albidas
(Cryptococcus albidus) IFO 0378, Cryptococcus terreus (Cryptococcusterreus) IFO 0727, Pichia farinosa IFO 0574, Pichia membranaefaciens
IFO 0460, Rhodosporium toluroides
sporidium toruloides) IFO 0871, Rhodotorula graminis IFO 0190, Rhodotorula minuta IFO 0387, Rhodetorula rubra IFO 0383, Saccharomyces IFO 0vis
0614, Trigonopsis variabilis
riabilis) IFO 0671.