JPS6265687A - Production of optically active (s)-2-halo-1-phenylethanol - Google Patents

Abstract

PURPOSE:To obtain the titled ethanol compound suitable as a synthetic raw material for pharmaceuticals, agricultural chemicals, animal drugs, perfumes, etc., easily in high efficiency, by contacting a haloacetophenone with a specific microorganism. CONSTITUTION:The objective phenylethanol of formula II can be produced by contacting (A) an alpha-haloacetophenone of formula I (X is halogen atom) with (B) a microbial strain capable of asymmetrically reducing the compound of formula I to the R-2-halo-1-phenylethanol of formula II having R-configuration and belonging to Ambrosiozyma genus, Arthroascus genus, Botryoascus genus, Citromyces genus, Filobasidium genus, Hanseniaspora genus, Hormoascus genus, Kloeckera genus, Lipomyces genus, Pichia genus, Actinomucor genus, Chlamydomyces genus, Mortierella genus, etc.

Description

Detailed Description of the Invention [Industrial Field of Application 1] The present invention provides α-haloacetophenone represented by the general formula [I] (wherein, X represents a halogen atom) as a general Formula [1] (wherein,
genus Arthroascus, genus Potulioascus, genus Sitelomyces, genus Filobacidium, genus Guilliermondella, genus Hanzeniasbora, genus Hormoascus, genus Chloerachella, genus Lipomyces, genus Kluyveromyces, genus Picya, genus Stefanoascus, Schizosatucharomyces, Trigonopsis, Absidia, Actytumcor, Baraxeera, Basidiolus, Chlamydomyces, Circinella, Coriolus, Helicostelum, Rangetes, Microthecium, Morteierella, Mucor, Neurospora, Prototheca,
Optical activity (R) characterized by contacting microorganisms selected from microorganisms belonging to the genus Fuicinoporus and Rhizolapus and collecting the produced (R)-2-halo-1-phenylethanol represented by the general formula circle. -2-no10-1
- This relates to a method for producing phenylethanol.

Optically active (R)-2-halo l-phenylethanol has two types of functional groups and is highly reactive (as it can be easily converted into R1-styrene oxide, it is useful for pharmaceuticals and animals that require optical activity). It is an extremely useful substance as a synthetic raw material for medicines, pesticides, fragrances, etc.

[Prior art and problems] Regarding the production method of (R)-2-halo-1-phenylethanol, D. Ridley et al. [Journal of Chemical
Society Chemical Communication (J,
Chem, Soc, Chem, Comm, ).

400, 1975], α-chloroacetophenone was synthesized from (R)-2- by Satucharomyces cerevisiae.
For chloro-1-phenylethanol, Mata H, Ziffer et al. [Journal of Organic Chemistry (
J, Org, Chem,), volume 45, page 8852,
[1980] reported that α-bromoacetophenone was converted to (R)-2-fromo1- by Cryptococcus macerans.
Add α-chloroacetophenone to phenylethanol (
reported that it was converted to R)-2-chloro-1-phenylacetophenone. However, these methods using microorganisms have weak reducing ability, require a large amount of yeast, and require a long time for reaction, so they are not advantageous for industrial implementation.

[Means for Solving Problems] Therefore, as a result of searching for microorganisms with strong reducing ability, the present inventors found that various microorganisms other than the microorganisms known to have the above-mentioned reducing ability produce α-haloacetophenone. Asymmetric reduction,
It was found that R1-2-halo-l-phenylethanol was produced with high efficiency.

Microorganisms that can asymmetrically convert α-haloacetophenone to R)-2-halo-1-phenylethanol used in the present invention can be found by the method described below.

For example, 40g glucose, 8f yeast extract,
(NH4)2HP018 f, KH2PO479
, MgSO4・7H200,8j', ZnSO4"
1112060 "%', FeSO4・7H20901
1y, CuSO4・5H205'I, MnSO4・4
After sterilizing 800 g of A medium with the composition of H2010''I, NaCl O, 1f (11 parts) in a 21-volume sloped flask, microorganisms were planted and cultured with shaking at 80°C for 2 days. Centrifugation Nyolose is suspended in water 75+w/containing 5%, and shaken at 80°C for 2 to 8 days in a 21-Sakaro flask.After that, an equal amount of ethyl acetate is added to perform extraction to produce 2-chloro-1. - phenylethanol by gas chromatography (column: silicon 0
v-17, -〇, 8X200 (11, column temperature 185
'C.

Analyze with N2 gas pressure 1.2 & 9/d). On the other hand, the optical purity of α-chloro-1-phenylethanol is determined by distillation and purification of the extracted oil, followed by high performance liquid chromatography (column:
Chiralcel-QC manufactured by JASCO Corporation.

Elution solvent hexane-ether (80:1), flow rate 2.2
g//*, detection at 220 nm), the fR1 body was detected at 29
The (S) form is separated with a retention time of 88 minutes and the optical purity can be determined. Also, microorganisms that can convert α-chloroacetophenone to (R)-2-chloro-1-phenylethanol are similar to α-promacetophenone (R).
)-2-bromo-1-phenylethanol.

The vine microorganism used in the present invention is Ambrosiozymaphile
ntoma) I F 0 1847, Arthroascus ja
vanensis ) IFO1848, Botryoascus sinnaedendrus (Botryoascus 5
ynnaedendrus ) IFO1604, Citeromyces manritinsis (Citeromyces
matritensis) I F 00651,
Filobasidium capsuligenus (Filobas)
idium capsuligenus ) I F
01119, Guylier Mondella Serenospora (
Guilliermondella 5elenosp
ora) I F 01850, Hanseniaspora
Barbiensis (Hanseniaspora va)
byensis) IFO1758, Hormoascus pladiodis (Hormoascus platy
odia) I F O1471, Kroetsukela'
Africana (Kloeckera africana)
) IFo 0869, l? Mysse Stuckei (
Lipomyces 5tarkeyi) I F
0 0678, Kluyveromyces fragilis (Kl
uyvercxrycesfrag 1s) IFO05
41. Pich
ia membranaefaciens )IFo
0189, Stefanoaskas Ciferrii (5te
phanoascus ciferrii) I F
O1854, Schizosaccharomyces bombe (Schi
zosaccharomyces pombe)
IF 0085B,) Lygonopsis palabilis (
Trigonopsls variabilis )
IFo 0671, Absidia corymbifera (
Absldiacorymbifera ) I FO
40f O, Actum Fool Elegance (Act
inomucor elegans) IFo 402
B. Backusella circina
circina) I Fo 916 B, Basidiobolus meristosporus
us) IFO9168, Gramydomyces palmarum (Ch Iamydomyces palmarum)
IFO8861, Circinella simplex (C1r
icinella simplex) IFO6412
, Coriolus fersicolor (Coriolus v.
ersicolor) I FO80888, Helicostylum nigricans
igricans) I F O8091, Lonzites betulina
) IFo 496B, Microthecium compressum
) I F O80249, Mortierella 1sabellina
) IFO7884, Mucor Javanicus (81uc
or javanicus) I F 0 4572
, Neurospora c.
rassa ) IFO6067, Prototheca zopfii IFO
6998, Pycinobolus cocineus (Pycnop
oruscocclneus) I F 0 649
6. Rhizolapus primar (Rh)
lemar ) IFo 4697, etc.

Any nutrient source that can be assimilated by these microorganisms can be used as a culture medium for these microorganisms. For example, carbohydrates such as glucose and sucrose; alcohols such as ethanol and glycerol; hydrocarbons such as paraffin; organic acids such as acetic acid and propionic acid; carbon sources such as soybean oil; or mixtures thereof; yeast extract, peptone, Nitrogen-containing inorganic organic nutrients such as meat extract, cornstap liquor, ammonium sulfate, ammonia; phosphates, magnesium, iron,
A conventional medium containing an appropriate amount of inorganic nutrients such as manganese and potassium; and vitamins such as biotin and thiamine is used. The culture method is to adjust the pH of the nutrient medium to 4.0-9.
Incubate aerobically at 20-40°C for 1-5 days.

The reduction reaction can be carried out using the culture solution as it is, or by separating the bacterial cells by centrifugation, resuspending them in a phosphate buffer or water, and adding α-haloacetophenone for one reaction. There are ways to do this. During this reaction,
Carbon sources such as glucose and sucrose may be added as energy sources. Furthermore, the bacterial cells may be kept as viable cells, or may be treated with acetone, freeze-drying, or the like. Furthermore, these bacterial cells can also be used by immobilizing them on a carrier.

The α-haloacetophenone may be added at tt of 1, or may be dissolved in an organic solvent that does not affect the reaction and added all at once or in portions from the beginning of the reaction. The reaction is carried out at a temperature of 10 to 60 °C at a pH of 5 to 9 and a temperature of 8 to 120 °C.
Carry out under stirring for hours.

The (R1-2-halo-1-phenylethanol produced by the reaction can be collected directly from the reaction solution or after separating one substance, extracted with a solvent such as ethyl acetate or dichloromethane, dehydrated, and then distilled to obtain highly pure (R1-2-halo-1-phenylethanol). ) -2-halo-1-phenylethanol is obtained with 6 easy IC.

〔Example〕

The present invention will be specifically explained below with reference to Examples, but the present invention is not limited to these Examples. The indicated microorganisms were inoculated. and 8
The culture was carried out aerobically for 2 days at 0°C with shaking. Bacterial cells were collected from this culture solution by centrifugation, suspended in 75 g of 0.1M phosphate buffer (pH 7.0) containing 0.5% α-chloroacetophenone and 5% sucrose, and placed in a 21-saka flask. A shaking reaction was performed at 80°C for 120 hours. After the reaction, the reaction solution was extracted with equal amounts of ethyl acetate (twice) to extract (R1-2-chloro-1-phenylethanol), and the ethyl acetate layer was analyzed by gas chromatography to examine the reaction rate. After dehydrating ethyl acetate with anhydrous sodium sulfate, the solvent was removed to obtain an oily substance. This was purified by mini-distillation to obtain (R)-2-chloro-1-phenylethanol. This was dissolved in methanol. The optical purity was measured using high performance liquid chromatography.The results are shown in Table 1.

Table 1 Continued from Table 1 Example 2 The microorganisms shown in Table 2 were cultured in the same manner as in Example 1, and α-bromoacetophenone was used instead of α-chloroacetophenone, and the reaction and analysis were conducted in the same manner. Table 2 shows the results.
Shown below.

Table 2 Example 8 A 51-volume mini jar containing A medium 31 was inoculated with Zeniaspora barbiensis IFO1758 and cultured for 24 hours at 80°C, aeration at 1 vvm, and stirring at 500 rpm. After culturing, the bacterial cells were collected by centrifugation, suspended in 750 ml of water, 7.5 f of α-chloroacetophenone and 88 g of sucrose were added, and the pH was adjusted to N.
Stir at 150 rp at 80°C while maintaining the temperature at 7.0 with aOH.
The reaction was carried out for 72 hours at m. After the reaction was completed, the mixture was extracted twice with 750 g/ethyl acetate. Ethyl acetate I! After dehydrating I with anhydrous sodium sulfate, the solvent was removed under reduced pressure to obtain an oily substance 8.21.

Distill this (75-76°C/21111H9) L,
6.2f of (R)-2-chloro-1-phenylethanol was obtained as a colorless oil. Its specific optical rotation is [α]25-5
2.8°■) (c=2.cyclohexane), and the optical purity was 100% e, e, according to high performance liquid chromatography analysis.

Example 4 Hanseniusbora barbiensis IF0 1758 was cultured in the same manner as in Example 8, and reacted with α-bromoacetophenone as a substrate. , / 0
．． 5 HL) was obtained. This specific optical rotation is [αID
49.0 (c-=1.rioform), and the optical purity as determined by high performance liquid chromatography analysis is 100.
%e, e.

[Effect of the invention 1 According to the present invention, as shown in Examples, optical activity (R1-
2-halo-1-phenylethanol can be produced efficiently.

Claims (3)

[Claims] (1) General formula [I] ▲There are mathematical formulas, chemical formulas, tables, etc.▼[I] (In the formula, X represents a halogen atom) Formula [II] ▲There are mathematical formulas, chemical formulas, tables, etc.▼ [II] (In the formula, X is the same as above) Asymmetrically reduced to (R)-2-halo-1-phenylethanol Ambrosiodymma, which has the ability to
Arthroascus, Botryoascus, Sitelomyces, Filobacidium, Guyilliermondella, Hanseniaspora, Hormoascus, Kloetschella, Lipomyces, Kluyveromyces, Pichia, Stephanoasca Schizosatucharomyces, Trigonopsis, Absidia, Acteinomucor, Batuxeella, Basideoborus, Chlamydomyces, Circinella, Coriolus, Helicosterum, Rangetes, Microthecium, Morteierella, Mucor Genus, Neurospora, Prototheca,
An optical system characterized in that the (R)-2-halo-1-phenylethanol produced by the general formula [II] is collected by contacting a microorganism selected from the microorganism group belonging to the genus Fuicinoporus and the genus Rhizotpus. Active (R)-2-halo-
Method for producing 1-phenylethanol. (2) The production method according to claim 1, wherein in the general formulas [I] and [II], the halogen atom is Cl or Br. (3) Microorganisms such as Ambrosiodymma florentoma, Arthroascus jabanensis, Botryoascus
Cinnaedendrus, Cytelomyces maturitensis, Filobacidium capsurigenus, Guyilielmondella selenospora, Hanseniaspora barbiensis, Phormoascus plateioides, Chloetsuchella africana, Lipomyces stucaei, Kluyveromyces fragilis, Pichia・Membrane aefaciens, Stephanoascus schifferii, Schizosatucharomyces pombe, Trigonopsis palabilis, Absidia corimbihuela, Acteinomucor elegans, Batuxera circina, Basideiobolus melistosporus, Chlamydomyces palmarum, Circinella Simplex, Coriolus fuersicolor, Helicosterum nigricans, Rangetes vetulina, Microthesium complexium, Morteierella isabelina, Mucor jiavanicus,
3. The method of manufacturing according to claim 1 or 2, which is Neurospora cratusa, Prototheca zoffii, Fuicinoporus cocineus, and Rhizopus delimar.