JPS61205498A - Production of axial asymmetric compound - Google Patents

Abstract

PURPOSE:To produce (R)-binaphthol and (S)-binaphthol in large amount in separated state, by hydrolyzing one of (R)-isomer and (S)-isomer of acylated binaphthol derivative with a specific bacterial strain or yeast. CONSTITUTION:A mixture of an acylated derivative of (R)-1, 1'-bi-2-naphthol and an acylated derivative of (S)-1, 1'-bi-2-naphthol is treated with the bacterial cell of Bacillus genus or yeast cell of Saccharomyces genus capable of hydrolyzing only one of the above components, or their cultured liquid, or enzymes (or enzyme-containing substance) originated therefrom, to effect the hydrolysis of only one of the components. The acylated derivative of optically active 1, 1'-bi-2-naphthol and that of optically active 1, 1'-bi-2-naphthol can be produced by this process. The components are separated from each other and one of them is hydrolyzed. The kind of the hydrolyzed isomer [(R)-isomer or (S)- isomer] depends upon the kinds of enzymes or acyl groups.

Description

Detailed Description of the Invention [Industrial Field of Application] The present invention provides (R)-1,1'-bi-2-naphthol and (
S)-1,1'-bi-2-naphthol, e.g. from the racemate, enzymatic K (R) and/or (S
) Concerning the method of preparing the whole.

[Conventional technology]

Optically active binaphthol has been known as an optically active reagent. For example, a reducing agent consisting of lithium aluminum hydride and ethanol plus optically active binaphthol gives a high asymmetric vehicle [Journal of the American Chemical Society (J, Am, Chem, So.
c,)+101e3129+.

1979], it is recognized as a very important reagent for the synthesis of physiologically active substances with optical activity and related substances, which have been actively attempted in recent years, especially for the synthesis of prostaglandins [Journal of American・Chemical, Sosaya Tea (J.

Am, Chem, Seo, ) 1101, 5843
, 1979].

Furthermore, it is used stoichiometrically or catalytically as an organic titanium reagent or a catalyst for hydroboration in the synthesis of optically active substances [Herpetica chimica acta (He1)].
v, Chim, Acta) + 6 branches, 2485
゜1981: and Tetrahedron Letters (Tet
rabedron Letters) + 1879
+ 1977] and are widely used as important optically active reagents.

A method for enzymatically resolving a racemate into an optically active form is already known, but a method for enzymatically resolving a racemic form of an axially asymmetric compound such as binaphthol of the present invention into an optically active form is not known. Na-.

Conventionally, optically active binaphthol has been produced in its racemic form i1. ，
It was produced by converting it into 1'-binaphthyl-2,2'-diylhydrodiene phosphate, optically resolving this derivative using an optically active base such as cinchonidine, and then hydrolyzing it again. However, this method is complicated and difficult to produce the required high purity optically active substance in large quantities and at low cost.

[Problem that the invention seeks to solve]

The object of the present invention is to provide a method for producing optically active binaphthol by an enzymatic method, which does not have the drawbacks of the chemical methods described above.

[Means for solving problems]

As a result of repeated research to fully establish the above-mentioned production method, the present inventors found that bacteria belonging to the genus Bacillus and Saccharomyces spp.
The yeast belonging to
We discovered that an enzyme can be produced that selectively hydrolyzes one of the acylated derivatives of naphthol.

The present invention was completed based on this knowledge.

Therefore, the present invention relates to a method for producing optically active binaphthol, which comprises the steps of: (s)-1,1'-bi-2-naphthol acylated derivative and (s)-1,1'-bi-2-naphthol; A mixture of naphthol with an acylated derivative is treated with a bacterium of the genus Bacillus or a bacterium of the genus Saccharomyces that can selectively hydrolyze the ester bond of one derivative in the mixture.
), the acyl group of the derivative is removed by treating it with cells of yeast of the genus Yeast, a culture solution containing the same, or an enzyme-containing substance or enzyme derived from the bacteria or the yeast, and the optically active 1,1'- Bi-2-naphthol and the optically active acylated derivative of 1,1'-bi-2-naphthol are separated from each other, and if desired, the optically active acylated derivative of 1,1'-bi-2-naphthol is separated. Hydrolyzing to remove the acyl group and recovering (S)-1,,1'-bi-2-naphthol and/or (S)-1,1'-bi-2-naphthol. 6 [Specific Description] In the method of the present invention, an acylated derivative of (S)-1,1'-bi-2-naphthol and (S)-1,1'-bi-2
- Mixtures of naphthol with acylated derivatives are used. These compounds are represented by the following formula.

However, the ratio of (S)integrate and (S)integrate can be variously different, but a racemic compound, which is an equal compound of these, is generally used. Various acyl groups can be used, such as acetyl group, glopionyl group, butyryl group,
Examples include benzoyl groups, and acylated products having these groups are 1,1'-binaphthyl-2,2, respectively.
'-diol diacetate), 1,1'-binaphthyl-2
, 2'-diol dipropionate, 1,1'-binaphthyl-2,2'-diol dibutyrate, and 1,1'-binaphthyl-2,2'-diol dipenzoate.

Whether the compound to be hydrolyzed in the method of the present invention is (R) monomer or (S) monomer differs depending on the strain used and the type of acyl group. For example, when using the L-75 strain,
When the substrate is an acetyl derivative, (S) is selectively hydrolyzed, whereas when the substrate is an n-butyryl derivative, (8) is selectively hydrolyzed.

The enzyme used in the method of the present invention is (lR)-1
, acylated derivatives of 1'-bi-2-naphthol and (S
)-1, 1'-Bi-2-naphthol acylated derivatives. This enzyme is a hydrolase that can selectively hydrolyze the ester bond of any of the acylated derivatives of acylated derivatives of 1,1'-bi-2-naphthol. Produced by yeast of the genus Satucharomyces. Examples of the bacteria of the genus Pacillus that produce the enzyme include Bacillus subtilis var.
1μ±) (IAM1633: ATCC' 9672
), Bacillus subtilis (IAM 1026:
ATCC9466), etc., and examples of yeasts of the genus Saecharomyces include, for example, Saecharomyces rouxii (Saecharomyces rouxii).
(IAM4962:ATCC2623), Saccharomyces ros
ei) (IAM-4991), etc.

These bacteria can be easily obtained by those skilled in the art from the above-mentioned depository institutions.

There are also microorganisms that produce the enzymes mentioned above. It can be easily separated from nature using conventional methods. For example, a microorganism is isolated from a source such as soil according to a conventional method, cultured in a medium in which the microorganism can grow, and the acetyl ester of carcinoid 1,1'-p-2-naphthol is added to the culture solution. dimethylformamide solution is added and inked under the same conditions as culture, and the reaction solution is analyzed by liquid chromatography, for example, to select microorganisms that selectively hydrolyze one of the optically active forms. do.

By such a method, it was found that about 10 strains of the isolated bacteria produced all of the above-mentioned enzymes. Therefore, those skilled in the art can easily obtain microorganisms having such activity from among preserved strains or from nature according to the disclosure of the present invention.

Examples of microorganisms isolated from the natural world in this way include 1. For example, Pacilrus s. and p.
, L-'75 strain.

This strain has the following mycological properties, namely:

(1) It is a bacillus: (R) It is a Durham-positive bacterium: (3) It has endospores: (4) It has pen hairs and is motile (migratory); Manual Op Determinative Bacteriology (Bergey & M
annual of DeterminativeBaC
It is classified into the genus Pasillas according to t@riology).

In culturing any of the above-mentioned microorganisms in the present invention, any conventional culture medium in which the microorganism to be cultured can grow can be used.

Either solid culture or liquid culture can be used as the culture method, but in order to implement it on a large scale, it is recommended to perform liquid culture under aerobic conditions, such as by aeration and stirring, or by taking pictures. Preferred layer.

As the culture temperature, any temperature within the range at which the specific microorganism to be cultured can be selected can be selected. For example, for the above-mentioned microorganisms, the temperature is 20°C to 38°C, preferably 30°C.
The three-incubation time that can be cultured at ~35°C varies depending on the microorganism, but is usually 12 to 48 hours.

The hydrolase of the present invention can be used in various forms, such as a culture solution containing cultured cells obtained by culturing the microorganism, cells isolated from the culture solution, Enzyme-containing liquid after the bacterial cells have been removed, crushed bacterial cells,
Partially purified enzyme preparations that have been purified to various stages can be used. Isolated and purified enzymes can also be used.

Further, it is also possible to use immobilized bacterial cells or immobilized enzymes in which these enzyme-containing substances or enzymes are immobilized according to a conventional method.

In the reaction, the enzyme or enzyme-containing substance is brought into contact with a mixture of (R) monomer and (S) monomer as raw materials in an entirely aqueous medium. As this aqueous medium, for example, water, a culture medium, or a buffer such as a phosphate buffer can be used. The reaction can be carried out under normal physiological conditions, for example at a temperature in the range of about 20-40°C, -about 5-8°C. The reaction time varies depending on the enzyme concentration in the reaction medium, raw material concentration, etc., but is approximately 0.5 to 12 days.

The concentration of the raw materials added varies depending on the enzyme titer in the reaction medium, etc., but is between 0.1 and 1. Ow/v 4 is convenient, preferably O13-0.4 w/v4. The raw materials can be added all at once, or can be added in multiple portions or continuously as the reaction progresses. The raw material can be added to the reaction medium in powder form, but if the concentration added is higher than the concentration dissolved in the reaction medium, the raw material can be added to the reaction medium at a high concentration in a suitable solvent in which the raw material can be dissolved at a high concentration, such as dimethylformamide. It is convenient to dissolve this solution in the reaction medium and add this solution to the reaction medium.

As described above, the reaction can be carried out in various manners, but from an industrial standpoint, it is convenient to carry out the following manner. That is, any one of the microorganisms of the present invention is inoculated into a liquid medium and cultured with aeration under the above conditions. After the bacteria have grown sufficiently, an acylated derivative of racemic 1,1'-p-2-naphthol is added, and the reaction is carried out under the same culture conditions as described above. This reaction is carried out with ventilation stopped, as the case may be.

After carrying out the reaction as described above, the optically active can)- or (S)-binaphthol in the reaction solution and (S)- or (8)-
)-acylated derivatives of binaphthol are separated from each other. This separation and purification can be carried out by any conventional method, such as column chromatography, solvent extraction, preparative thin layer chromatography, etc. The separated (8)- or (S)-binaphthol is then purified to obtain the final product. On the other hand, the unreacted acylated derivative of optically active (S)- or (phase)-binaphthol is deacylated by a conventional hydrolysis method to obtain optically active (S)- or (phase-binaphthol).

〔Example〕

Next, this invention will be explained in more detail with reference to Examples.

Example 1゜2$-) IJ Endo Broth (NutrlentBro
th) (trade name, manufactured by Difco) 20d with Na2H
The mixture was adjusted to pH 7.0 using PO4, placed in a 100-mm triangular container, and sterilized. In this medium, Pacilrus subtilis variety niger (IAN1633:
ATCC9672) was inoculated and cultured with shaking at 30 to 35°C for 2 days.

Racemic 1,1'-binaphthyl-2,2'-diol diacetate) 20! t-dimethylformamide 0.6dK
Dissolve and add this solution to the above culture solution.

Subsequently, the mixture was shaken and reacted at 30 to 35°C for 11 days.

After the reaction was completed, the reaction solution was extracted with ethyl acetate and subjected to preparative thin layer chromatography 4 (Merek F524 #20
1, manufactured by Merck & Co.) (Developing solvent is benzene: ethyl acetate 1)
00: IT use) Busy 1 separated and 9 unresolved 1,1'
-binaphthyl-212'-diol diacetate (8,
The optical purity of 5ffil;I) and hydrolyzed 1,1'-b-naphthol (7 mg) was measured as follows.

1.1'-binaphthyl-2,2'-diol diacetate was hydrolyzed using 51KOH-methanol solution at 25°C for 30 minutes, and the resulting 1,11-bi-2-naphthol was dissolved in pyridine. Treated with (+)-α-methoxyα-trifluoromethyl-α-phenylacetic acid chloride (pnph-ct) at 25°C for 12 hours,
It was made into a diester of MTPA. Add this to the liquid.
bax SIL column, 15 cm x 4.6 sm, manufactured by Shimadzu Tennis Co., Ltd.). Hexane-methylene chloride 4:1 was used as the solvent system, flow rate was 3 g/min, 24
Detection was performed by absorbance at 0 nm. Known samples as standard products (R)-1,1'-bi-2-naphthol and (
Each MTPA ester of S)-1, 1'-bi-2-naphthol was used. The peak times of (8) and (S) were 18.0 minutes and 15.0 minutes, respectively.

The optical purity of this product was approximately 54 mm. Enzymatically hydrolyzed 1,II-bis-2-7toll was analyzed in the same manner (however, 5-KOT (without hydrolysis with methanol, IA)), and the optical purity was 49. I regret it.

Example 2 A medium was prepared by dissolving 120 μl of yeast extract, 200 μl of pezoton, and 200 μl of gluyose in 20 ml of purified water, and the strains S. s., p., and L-75 were used as the bacterial strains. Racemic 1,1'-binaphthyl- as a substrate
2,2'-diol diacetate 801n9'i was used. The (S) monomer was selectively hydrolyzed, and the optical purity of the diacetate mainly composed of the (8) monomer was about 701.

Example 3 The method of Example 1 was followed. However, Pacilrus subtilis s, p, L-75 was used as the bacterial strain, and racemic 1,1'-binaphthyl-2,2'-dioluci n-butyrate was used as the substrate. sail) was selectively hydrolyzed.

Example 4 The method of Example 1 was repeated. However, various strains were used. The conversion yield and optical purity for each strain were as follows.

Procedural Amendment (Spontaneous) Date: July 71, 1985 Manabu Shiga, Commissioner of the Japan Patent Office1, Indication of the case: Patent Application No. 45755, filed in 19852, Title of the invention: Process for producing axially asymmetric compounds 3, Amended sound Patent applicant name (190) Suntory Ltd. 4, agent address Shizumitsu Toranomon Building, 8-1o Toranomon-chome, Minato-ku, Tokyo 105 Phone number (504) 0721 Name Patent attorney (65)
79) Akira Aoki (4 others) 5. Specification to be amended 6. Contents of amendment (no change in content) 7. Catalog-type written specification of attached documents 1 written procedural amendment (voluntary) Showa June 7, 1961 Michibe Uga, Commissioner of the Patent Office1, Indication of the case, Patent Application No. 45755 of 19852, Name of the invention, Process for producing axially asymmetric compounds3, Relationship with the amended person case Patent applicant name (190) Suntory Ltd. 4, agent address 5-8-10 Toranomon-chome, Minato-ku, Tokyo 105
Subject of amendment (1) “Claims” column of the specification.

(R) "Detailed Description of the Invention" column of the specification.

6. Contents of correction (1) Amend the claims as per the attached sheet.

(R)■Specification (typed specification based on procedural amendment submitted on April 18, 1985, the same applies hereinafter) page 7, line 3 rL-75 strain'' rThe inventors have submitted the specification in Adachi, Tokyo. Bacillus strain L-75 (its properties will be described later) isolated from the soil in the district.

■Page 9, line 6, "Meguro-ku" is corrected to "Adachi-ku".

■Page 15, line 9, "Bacillus" is corrected to "Bacillus".

■Page 15, lines 16 to 20 (contents of Example 3)
is corrected as follows.

'Uni) Nutrient Bro
Th.

pFI adjusted to 7.0 with NatHPOa) 100ml
In, Bacillus subtilis 3. p, L, 75 were inoculated and cultured with shaking at 30-35°C for 2 days. To this, 100 μ of 1,1'-binaphthyl-2,2'-
A solution of diol-di-robethyrate dissolved in 3 ml of dimethylformamide was added to the above culture solution,
The reaction was carried out by shaking at 35°C for 122 days.

The reaction solution was extracted with ethyl acetate, and the extract was separated and purified using liquid chromatography to obtain the (R)-diol form with an optical purity of 91% and the (S)-diacetate form with an optical purity of 98%. Ta. The yield was 92%. ” ■The following statement is added after Example 4 on page 16.

Using a commercially available pig liver-derived esterase (purchased from Sigma) as an asymmetric hydrolase, a racemic 1.1'-binaphthyl-2,2'-diol- of an axially asymmetric compound was prepared. Although a hydrolysis reaction using diacetate and racemic 1,1'-binaphthyl-2,2'-diol-dipropionate as substrates was attempted, no optically active diol or monoacetate was obtained. 2. Claim 1: A mixture of an acylated derivative of (R)-1,1'-bi-2-naphthol and an acylated derivative of (S)-1,1'-bi-2-naphthol. , bacteria of the genus Bacillus or yeast of the genus Saccharomyces that can selectively hydrolyze the ester bond of one of the derivatives in the mixture;
Or, the acyl group of the derivative is removed by treating it with a culture solution containing it, or an enzyme-containing substance or enzyme derived from the bacteria or the yeast, and the optical activity is 1,1.
'-B-2-naphthol and optical activity m1,1 '- and -2
- acylated derivatives of naphthol, and if desired, the optically active acylated derivatives of 1,1'-bi-2-naphthol are hydrolyzed to remove the acyl group, and (R)-1, 1'-bi-2-naphthol and/or (
(R)-1,1'-bi-2-naphthol and/or (S)-1,1'-bi-2, characterized in that S)-1,1'-bi-2-naphthol is recovered. - A method for producing naphthol.

Claims (1)

[Claims] 1. Into a mixture of an acylated derivative of (R)-1,1'-bi-2-naphthol and an acylated derivative of (S)-1,1'-bi-2-naphthol, one of the acylated derivatives in the mixture is added. A bacterial cell of a bacterium of the genus Bacillus or a yeast of the genus Saccharomyces that can selectively hydrolyze the ester bond of a derivative of , or a culture solution containing the same, or an enzyme derived from the bacterium or the yeast. The acyl group of the derivative is removed by the action of a compound or an enzyme, and the optically active 1,1'-
Bi-2-naphthol and the optically active acylated derivative of 1,1'-bi-naphthol are separated from each other, and if desired, the optically active acylated derivative of 1,1'-bi-2-naphthol is hydrolyzed. to remove the acyl group and (R)-
1,1'-bi-2-naphthol and/or (S)-1,
(R)-1,1'-bi-2-naphthol and/or (S) characterized in that 1'-bi-2-naphthol is recovered.
)-1,1'-bi-2-naphthol production method.