JPH10304885A - Production of optically active alcohol compound - Google Patents

Abstract

PROBLEM TO BE SOLVED: To readily produce the subject compound used for cosmetics, pharmaceuticals, etc., by allowing a hydrolytic enzyme of (2R, 3S) isomer of an N-acylsphingoid ester to act on the ester, recovering the unreacted material, and hydrolyzing the recovered material. SOLUTION: An ester hydrolytic enzyme having activities for asymmetrically hydrolyze an ester bond part of (2R, 3S)-isomer of an N-acylsphingoid ester of formula I [R<1> and R<2> are each a 7-31C aliphatic hydrocarbon group (substituted with one or more hydroxyl groups); R<3> and R<4> are each a 2-7C acyl group] is allowed to act on the N-acylsphingoid ester or formula I to asymmetrically hydrolyze the N-acylsphingoid ester, and the unreacted (2S, 3R) isomer is recovered from the reaction mixture. Further, the recovered unreacted isomer is subjected to an ester hydrolysis to provide the objective optically active alcohol compound of formula II (one of R<5> and R<6> is H and the other is H or a 2-7C acyl) in the method for producing the optically active alcohol compound. The optically active compound is useful as cosmetics, pharmaceuticals, etc., and is safely produced without complex operation.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

[0001] The present invention relates to a method for producing an optically active alcohol compound.

[0002]

[Prior Art] General formula 3

[0003]

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[0004] (wherein, R ₁ and R ₂ are the same or different, represent 1 31 from seven carbon atoms which may be substituted with one or more hydroxyl groups of an aliphatic hydrocarbon, R ₅ And R ₆ at least one of them represents a hydrogen atom, and the other represents a hydrogen atom or an acyl group having 2 to 7 carbon atoms.) Useful as cosmetics and pharmaceuticals or intermediates for producing them. These compounds are usually extracted and separated mainly from epidermal tissues of animals such as cattle and pigs, but have problems such as limited production and difficulty in stable supply. Recently, an asymmetric synthesis method of an optically active sphingoid by an organic synthetic chemistry technique has been reported, and the optically active sphingoid can be converted to N-sphingoid using N-succimidyl octadecanoate, for example.
It is also known to lead to acyl sphingoids [R. Ju
lina et al., Helvetica Chemica Acta, 69, p. 368 (198
6)]. As an asymmetric synthesis method of optically active sphingoids by such an organic synthetic chemistry technique, for example, the chemical formula 4
An arpless asymmetric epoxidation is used to synthesize an epoxy compound represented by the formula (5), and an optically active formula (7) is obtained via the oxazolidinone compound represented by the formula (6).
R.Julina et al., Helevetica Chimica Acta, 69, p.368 (19
86).

[0005]

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[0006]

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[0007]

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[0008]

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Also, starting from Z-butene-1,4-diol as a starting material, Sharpless oxidation, epoxide cleavage with azide, oxidation to aldehyde, Wittig reaction, photoisomerization, and optically active formula H. Shibuya et al., Tetrahedron Letters, 30, p. 7205
(1989), etc., the method using Evans chiral auxiliary
Icolaou et al., J. Am. Chem. Soc., 110, p. 7910 (1988) et al., asymmetric hydrogenation of 2-N-acylamino-higher acyl acetate compounds with a ruthenium-optically active phosphine complex as a catalyst. A method for controlling asymmetric carbon by using the method is described in JP-A-6-80617 and the like.

[0010]

However, the production methods using these organic synthetic chemistry techniques are industrially carried out in view of the complexity of the operation, the number of required steps, the safety of the reaction materials and the amount used, and the like. However, it is not always a satisfactory method.

[0011]

Under such circumstances, the present inventors have developed an optically active compound represented by the above formula 3 which is simpler, safer, and more suitable for industrial scale implementation. As a result of intensive studies on a method for producing an alcohol compound, a biochemical method using an ester hydrolase having a specific ability was found, and the present invention was reached. That is, the present invention provides: 1) General formula 8

[0012]

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(Wherein R ₁ and R ₂ are the same or different and each represents an aliphatic hydrocarbon group having 7 to 31 carbon atoms which may be substituted with one or more hydroxyl groups; _Three
And R ₄ are the same or different and represent an acyl group having 2 to 7 carbon atoms. ), An ester hydrolase capable of asymmetrically hydrolyzing the ester bond site of the (2R, 3S) form of the N-acylsphingoid ester acting on the N-acylsphingoid ester represented by the general formula 8 After asymmetric hydrolysis, the unreacted (2S, 3R) -form of the N-acyl sphingoid ester is recovered from the reaction solution, and this is subjected to ester hydrolysis to give a compound of the general formula 9

[0014]

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(Wherein R ₁ and R ₂ are the same or different and each represents an aliphatic hydrocarbon group having 7 to 31 carbon atoms which may be substituted with one or more hydroxyl groups; _Five
And R ₆ , at least one of which represents a hydrogen atom, and the other represents a hydrogen atom or an acyl group having 2 to 7 carbon atoms. A) a method for producing an optically active alcohol compound characterized by obtaining an optically active alcohol compound represented by the following formula (hereinafter referred to as the production method of the present invention): 2) an N-acyl sphingoid ester represented by the general formula: 3. The method for producing an optically active alcohol compound according to the above 1, which is an erythro form, 3) the ester hydrolase has an amino acid sequence represented by SEQ ID NO: 1 or one or more amino acids in the amino acid sequence added, deleted or substituted 3. The method for producing an optically active alcohol compound according to the above item 1 or 2, which is a protein having the sequence described above.

[0016]

BEST MODE FOR CARRYING OUT THE INVENTION Hereinafter, the present invention will be described in detail.
General formula 1 used as a raw material of the production method of the present invention
0

[0017]

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(Wherein R ₁ and R ₂ are the same or different and each represents an aliphatic hydrocarbon group having 7 to 31 carbon atoms which may be substituted with one or more hydroxyl groups; _Three
And R ₄ are the same or different and represent an acyl group having 2 to 7 carbon atoms. In the N-acyl sphingoid ester represented by), as the aliphatic hydrocarbon group having 7 to 31 carbon atoms which may be substituted with one or more hydroxyl groups represented by R ₁ and R ₂ , For example, carbon atom 7 optionally substituted with one or more hydroxyl groups
A linear or branched alkyl group having 7 to 31 carbon atoms, having 1 or more double bonds and optionally substituted by one or more hydroxyl groups; Examples thereof include a branched alkenyl group. Preferably, a linear or branched alkyl group having 7 to 31 carbon atoms, which may be substituted with about 1 to about 3 hydroxyl groups, having about 1 to about 3 double bonds. And a linear or branched alkenyl group having 7 to 31 carbon atoms which may be substituted with about 1 to about 3 hydroxyl groups. Specifically, for example, heptyl group, tridecyl group, tetradecyl group, pentadecyl group,
Hexadecyl group, heptadecyl group, octadecyl group, nonadecyl group, eicosyl group, heneicosyl group, docosyl group, tricosyl group, tetracosyl group, pentacosyl group, hexacosyl group, heptacosyl group, octacosyl group, nonacosyl group, triacontyl group, hentriacontyl group, 1-tridecenyl group, 1-tetradecenyl group,
1-pentadecenyl group, 1-hexadecenyl group, 1-heptadecenyl group, 1-octadecenyl group, 1-nonadecenyl group, 1-eicosenyl group, 1-hydroxytridecyl group, 1-hydroxytetradecyl group, 1-hydroxypentadecyl group , 1-hydroxyhexadecyl group, 1-
Hydroxyheptadecyl group, 1-hydroxyoctadecyl group, 1-hydroxynonadecyl group, 1-hydroxyeicosyl group, 1-hydroxyheneicosyl group, 1-hydroxydocosyl group, 1-hydroxytricosyl group, 1
-Hydroxytetracosyl group, 1-hydroxypentacosyl group, 1-hydroxyhexacosyl group, 1-hydroxyheptacosyl group, 12-methyl-tridecyl group, 14
-Methyl-pentadecenyl group, methylheptadecyl group and the like. Carbon atom 2 represented by R ₃ and R ₄
From 7 to 7 acyl groups include, for example,

[0019]

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(Wherein, R ₇ represents an alkyl group having 1 to 6 carbon atoms). More specifically, for example, an acetyl group, a propionyl group, a butyryl group and the like can be mentioned. Preferably, for example, an acetyl group is used.

The N-acyl sphingoid ester represented by the general formula (10) can be obtained by converting the corresponding N-acyl sphingoid into, for example, T. Kolter et al., Tetrahedron, 50, p. 13425.
(1994) and the like, and can be produced by esterifying the compound.

Specific examples of such N-acyl sphingoid esters include, for example, N-eicosanoyl-
1,3-O-diacetyl-2-aminohexadecane-1,3-diol, N-eicosanoyl-1,3-O-diacetyl-2-aminoheptadecane-1,3-diol, N-eicosanoyl-1,3 -O-Diacetyl-2-amino-4-octadecene-1,3-diol, N-eicosanoyl-1,3-O-diacetyl-2-aminooctadecane
-1,3,4-triol, N-eicosanoyl-1,3-O-diacetyl-2-aminoeicosane-1,3-diol, N-eicosanoyl-1,3-O-dibutyryl-2-aminoeicosane -1,3-diol, N-eicosanoyl-1,3-O-diacetyl-2-aminopheneicosane-1,3-diol, N-octadecanoyl-1,3-O
-Diacetyl-2-amino-15-methylhexadecane-1,3-diol, N-octadecanoyl-1,3-O-diacetyl-2-
Aminodocosan-1,3-diol, N-octadecanoyl-1,3-O-dibutyryl-2-amino-4-hexadecene-1,3-diol, N-octadecanoyl-1,3-O-diacetyl- 2-amino-4-octadecene-1,3-diol, N-octadecanoyl-1,3-O-diacetyl-2-aminooctadecane-1,3,4-
Triol, N-octadecanoyl-1,3-O-diacetyl-
2-amino-4-heptadecene-1,3-diol, N-octadecanoyl-1,3-O-dibutyryl-2-amino-4-eicosene-1,
3-diol, N-2'-hydroxyoctadecanoyl-1,3
-O-Diacetyl-2-amino-15-methylhexadecane-1,3-
Diol, N-2'-hydroxyhexadecanoyl-1,3-O
-Diacetyl-2-aminohexadecane-1,3-diol, N
-2'-hydroxyhexadecanoyl-1,3-O-dibutyryl-
2-aminohexadecane-1,3-diol, N-2'-hydroxyeicosanoyl-1,3-O-diacetyl-2-amino-4-octadecene-1,3-diol, N-2'-hydroxyeico Sanoyl-1,3-O-diacetyl-2-aminooctadecane-1,
3,4-triol, N-2'-hydroxytetracosanoyl
-1,3-O-diacetyl-2-aminoeicosane-1,3-diol, N-2'-hydroxyhexacosanoyl-1,3-O-dibutyryl-2-aminoheptacosane-1,3-diol , N-docosanoyl-1,3-O-diacetyl-2-amino-15-methylhexadecane-1,3-diol, N-tecolacosanoyl-1,3-O-diacetyl-2-aminohexadecane-1,3-diol , N-hexacosanoyl-1,3-O-dibutyryl-2-aminohexadecane-1,3-diol, N-methyloctadecanoyl-1,3-O-
Diacetyl-2-amino-4-octadecene-1,3-diol,
N-pentacosanoyl-1,3-O-diacetyl-2-amino-4-
Docosene-1,3-diol, N-pentacosanoyl-1,3-O-
Diacetyl-2-amino-4-octadecene-1,3-diol,
Examples include, but are not limited to, N-pentacosanoyl-1,3-O-diacetyl-2-aminoeicosane-1,3-diol.

The N-acyl sphingoid ester represented by the general formula (I) used as a raw material in the production method of the present invention is preferably an erythro isomer, and its steric configuration is (2S, 3R) or (2R, An arbitrary mixture of isomers which are 3S) forms can be given.

The ester hydrolase that can be used in the production method of the present invention includes asymmetric hydrolysis of the ester bond site of the (2R, 3S) form of the N-acylsphingoid ester represented by the general formula (10). Any enzyme having the ability may be used. Ester hydrolases having such a capability are:
Esterases and proteases including lipases in a narrow sense, derived from animals such as pigs and humans, and from plants such as castor, Aspergillus, Candida, Fusarium, Geotricum, Mucor, Nocardia, Penicillium , Rhizopus, Saccharomyces, Achromobacter, Acinetobacter, Alcaligenes, Chromobacterium, Escherichia, Pseudomonas, Sphingomonas, Bacillus, Burkholderia, Morexella, Lactobacillus, Star It may be derived from a microorganism belonging to the genus Philococcus, Serratia, Yarrowia or the like. In addition, the genes for these ester hydrolases were isolated by recombinant DNA technology,
An ester hydrolase produced by a transformant obtained by introduction into a host cell belonging to the same genus or a host cell belonging to a different genus may be used.

Examples of more specific ester hydrolases include a protein having the amino acid sequence of SEQ ID NO: 1 or one or more amino acids in the amino acid sequence of SEQ ID NO: 1, which are added, deleted or substituted. And a protein having the ability to asymmetrically hydrolyze the ester bonding site of the (2R, 3S) form of the N-acylsphingoid ester represented by the general formula (10). The “protein having the amino acid sequence represented by SEQ ID NO: 1” is deposited, for example, under the Budapest Treaty as FERM-BP5740 with the Ministry of International Trade and Industry, National Institute of Advanced Industrial Science and Technology, Institute of Biotechnology and Industrial Technology (acceptance date: November 7, 1996) ) Microorganisms (E. coli JM109 / pAL612 strain). In addition, “a sequence having one or more amino acids added, deleted or substituted in the amino acid sequence represented by SEQ ID NO: 1;
(2) of an N-acyl sphingoid ester represented by
The “protein having the ability to asymmetrically hydrolyze the ester bond site of the (R, 3S) form” is, for example, a “cloning and sequencing” (Watanabe) Screening of the above animal, plant, or microbial gene library using the hybridization method described in Jikken Supervision, edited by Masahiro Sugiura, 1989, published by Rural Culture Co., Ltd. It can be prepared by isolating and expressing the resulting gene in a host cell such as a microorganism. Further, for example, a gene having the nucleotide sequence represented by SEQ ID NO: 2
lar Cloning, 2nd ed., Cold Spring Harbor Laborator
y Press (1989) et al.
It can also be prepared by introducing a mutation in which an amino acid is added, deleted or substituted, and expressing the obtained gene in a host cell such as a microorganism.

The ester hydrolase used in the production method of the present invention may be used in the reaction in the form of a culture of a microorganism or a cell containing itself, but the culture or the ester hydrolase containing the ester hydrolase may be used. The enzyme may be separated from the target tissue and used for the reaction in the form of a crude enzyme, a purified enzyme, or the like. Such a crude enzyme or a purified enzyme can be obtained, for example, by 1) sonication, 2) grinding using glass beads or alumina,
3) French press treatment, 4) Enzyme treatment such as lysozyme, 5) Warm blender treatment or the like to crush bacterial cells, cells or tissues, etc., and 6) Salting out using ammonium sulfate or the like from the obtained crushed product, 7) Precipitation with an organic solvent, an organic polymer such as polyethylene glycol, 8) various kinds of chromatography such as ion exchange chromatography, hydrophobic chromatography, gel filtration chromatography, affinity chromatography, and 9) preparation by a usual method such as electrophoresis. be able to. Furthermore, the ester hydrolase is insolubilized by a method of immobilization such as a carrier binding method in which the ester hydrolase is bound to the carrier by covalent bond, ionic bond, adsorption, or the like, or an entrapment method in which the ester hydrolase is confined in a polymer network structure. It can also be used in the form of a fixed object that has been processed to be easily separable.

The reaction temperature for the asymmetric hydrolysis is in the range of about 20 ° C. to about 70 ° C., preferably about 25 ° C. to about 40 ° C. The reaction is carried out according to the above-mentioned general formula.
It is preferable to use a two-layer system consisting of an organic solvent for dissolving the N-acyl sphingoid ester represented by the formula and a buffer solution for dissolving the ester hydrolase. If necessary, a surfactant may be added. Examples of the reaction solvent include decane, and examples of the buffer include ordinary buffers having a pH of about 5 to about 8. The reaction time is about 1
From time to about one week. When recovering the (2S, 3R) form of the N-acyl sphingoid ester from the reaction solution,
For example, methods such as solvent extraction, fractional distillation, and column chromatography can be used as appropriate. (2S, 3R) of the N-acyl sphingoid ester thus recovered
The optically active alcohol compound represented by the above general formula 9 can be obtained, for example, by subjecting the compound to alkaline hydrolysis by a usual method.

[0028]

EXAMPLES Hereinafter, the present invention will be described in more detail with reference to examples, but the present invention is not limited to these examples.

Example 1 A mixture of 10 ml of decane and 100 ml of 100 mM phosphate buffer (pH 7.1) was mixed with a racemic and erythro erythro-3 erythro-3.
1.03 g of -acetoxy-2-stearoylaminohexadecyl acetate and 50 mg of ester hydrolase prepared in Reference Example 1 described below were added, and the mixture was stirred and reacted at 30 ° C. The reaction was followed by TLC [silica gel, chloroform: methanol (15: 1)]. After reacting for 2 days, the reaction solution was extracted with ethyl acetate and then with chloroform. The collected organic solvent layer was washed with brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain a residue. The obtained residue was separated by silica gel columnography. Elution was performed with hexane: ethyl acetate (5:
1) followed by chloroform: methanol (30: 1). Eluted (+)-(2S, 3R) -erythro-3-acetoxy-
2-stearoylaminohexadecyl acetate (264.
After hydrolyzing a part of (9 mg), p-toluenesulfonic acid, 2,2-dimethoxypropane, and acetone (4R, 5
The product was converted to (S) -erythro-2,2-dimethyl-5-stearoylamino-4-tridecyl-1,3-dioxane and analyzed for optical purity by ¹ H-NMR. As a result, it was> 95% ee. Next eluted
After partially hydrolyzing (-)-(2R, 3S) -erythro-3-acetoxy-2-stearoylaminohexadecane-1-ol (481.8 mg), (-)-(2R, 3S) -erythro- As a result of comparison with the optical rotation of 2-stearoylamino-1,3-hexadecanediol, the optical purity was 16% ee. Further eluted,
A portion of (-)-(2R, 3S) -erythro-2-stearoylamino-1,3-hexadecanediol (214.8 mg) was added in the presence of p-toluenesulfonic acid, 2,2-dimethoxypropane, acetone,
(4S, 5R) -erythro-2,2-dimethyl-5-stearoylamino
It was converted to -4-tridecyl-1,3-dioxane, and the optical purity was analyzed by ¹ H-NMR. As a result, it was> 95% ee.

Example 2 A mixture of 0.1 ml of decane and 1.0 ml of a 100 mM phosphate buffer (pH 7.1) was added to a mixture of racemic and erythro erythro-3.
-Acetoxy-2-stearoylaminohexadecyl acetate (10.2 mg) and immobilized ester hydrolase (10.8 mg) prepared in Reference Example 2 were added, and the mixture was stirred and reacted at 30 ° C. The reaction was performed using TLC [silica gel, chloroform:
Methanol (15: 1)]. After the reaction for 2 days, the product (+)-(2S, 3R) -erythro-3-acetoxy-2-stearoylaminohexadecyl acetate was separated according to Example 1. After hydrolyzing a part of the separated acetate, it is reacted with α-methoxy-α-trifluoromethylphenylacetate (hereinafter referred to as MTPA) in the presence of dicyclohexylcarbodiimide and dimethylaminopyridine to form a bis-MTPA ester. After that, the optical purity was analyzed by ¹ H-NMR. (+)-(2S, 3R) -erythro-3-acetoxy-
2-Stearoylaminohexadecyl acetate was obtained in 50% yield and its optical purity was> 95% ee.

Reference Example 1: Preparation of ester hydrolase E. coli recombinant E. coli JM109 / pAL612 strain (FERM-BP 574)
0) was cultured in 100 ml of LB medium (manufactured by Difco Co.) containing 50 mg / L of ampicillin and 1 mM of isopropyl thio-β-D-galactoside at 37 ° C. for 16 hours, followed by centrifugation (6000 rpm, (10 minutes) to collect the cells. The collected cells were suspended in 10 ml of 100 mM phosphate buffer (pH 7.0), sonicated (10 minutes), and the crushed product was centrifuged to obtain a crude enzyme extract. .
The obtained crude enzyme extract was freeze-dried to obtain a crude enzyme powder.

Reference Example 2: Immobilization of ester hydrolase 0.1 M phosphate buffer (p) in which Triton X-100 (registered trademark of Union Carbide Chemicals Ant Plastics Co., Ltd.) (300 mg) was dissolved.
H7.0) To 10 ml of the enzyme powder obtained in Reference Example 1 was added 1 g, and then florisil [registered trademark of USSilica Company, sold by Sigma-Aldrich Japan KK] in ice water (8.7).
mg) was added. The resulting mixture was frozen at −78 ° C. and freeze-dried to obtain the immobilized powder enzyme (1.4
g) was obtained.

[0033]

According to the present invention, there is provided a method for producing an optically active alcohol compound represented by the above formula (3) by a biochemical method which is not complicated, does not require multiple steps, and is excellent in safety. I do.

[0034]

[Sequence list]

 SEQ ID NO: 1 Sequence length: 363 Sequence type: Amino acid Topology: Linear Sequence type: Protein Sequence Met Ser Arg Ser Ile Arg Ala Lys Ala Val Ala Thr Val Val Ala Ile 1 5 10 15 Ala Met Asn Ala Ala Pro Ala Ala Ser Val Gly Thr Val Leu Ser Leu 20 25 30 Ala Gly Ala Gln Ala Ala Ser Ala Ala Thr Thr Ala Val Asp Asp Tyr 35 40 45 Ala Ala Ala Thr Arg Tyr Pro Ile Ile Leu Val His Gly Leu Thr Gly Thr 50 55 60 Asp Lys Tyr Gly Gly Val Val Glu Tyr Trp Tyr Arg Ile Pro Glu Asp 65 70 75 80 Leu Arg Ala His Gly Ala Ala Val Tyr Val Ala Asn Leu Ser Gly Phe 85 90 95 Gln Ser Asp Asp Gly Pro Asn Gly Arg Gly Glu Gln Leu Leu Ala Phe 100 105 110 Val Lys Gln Val Leu Ala Ala Thr Gly Ala Gln Lys Val Asn Leu Ile 115 120 125 Gly His Ser Gln Gly Gly Leu Thr Ser Arg Tyr Val Ala Ser Val Ala 130 135 140 Pro Glu Leu Val Ala Ser Val Thr Thr Ile Ser Thr Pro His Trp Gly 145 150 155 160 Ser Gln Phe Ala Asp Phe Val Gln Gln Leu Leu Gln Thr Asp Pro Thr 165 170 175 Gly Leu Ser Ser Thr Val Leu Gl y Ala Phe Ala Asn Ala Leu Gly Thr 180 185 190 Leu Thr Ser Ser Asn Phe Asn Thr Asn Gln Asn Ala Ile Gln Ala Leu 195 200 205 Ser Val Leu Thr Thr Ala Lys Ala Ala Ala Tyr Asn Gln Lys Phe Pro 210 215 220 Ser Ala Gly Leu Gly Ala Pro Gly Ser Cys Gln Thr Gly Ala Pro Thr 225 230 235 240 Glu Thr Val Gly Gly Asn Thr His Leu Leu Tyr Ser Trp Gly Gly Thr 245 250 255 Ala Ile Gln Pro Thr Ala Thr Val Ala Gly Val Thr Gly Ala Val Asp 260 265 270 Thr Ser Val Ser Gly Val Thr Asp Pro Ala Asn Ala Leu Asp Pro Ser 275 280 285 Thr Leu Ala Leu Leu Gly Ser Gly Thr Val Met Ile Asn Arg Ser Ala 290 295 300 Gly Pro Asn Asp Gly Val Val Ser Gln Cys Ser Ala Arg Phe Gly Gln 305 310 315 320 Val Leu Gly Thr Tyr His Trp Asn His Thr Asp Ala Ile Asn Gln Ile 325 330 335 Leu Gly Val Leu Gly Ala Asn Val Glu Asp Pro Val Ala Val Ile Arg 340 345 350 Thr Asp Ala Asn Arg Leu Lys Leu Ala Gly Val 355 360 363

SEQ ID NO: 2 Sequence length: 1089 Sequence type: nucleic acid Number of strands: double-stranded Topology: linear Sequence type: Genomic DNA sequence ATG AGC AGA TCG ATA CGA GCG AAG GCA GTG GCG ACC GTG GTG GCG ATC 48 AAC GCG GCC CCG GCC GCG AGT GTT GGA ACC GTT CTG GCC ATG TCG CTG 96 GCC GGC GCA CAG GCC GCT TCC GCC GCG ACG ACC GCC GTT GAC GAC TAC 144 GCG GCG ACC CGG TAC CCG ATC ATT CTC GTG CAGGG CTG ACC GGC ACC 192 GAC AAG TAC GGT GGC GTC GTC GAG TAC TGG TAT CGC ATT CCG GAG GAC 240 CTG CGG GCG CAC GGC GCG GCG GTA GTA TAC GTT GCC AAC CTG TCC GGC TTC 288 CAG AGC GAC GAT GGC CCG AAC GGG CGT GGC CAA TTG CTT GCA TTC 336 GTG AAG CAG GTG CTC GCG GCG ACG GGC GCG CAG AAG GTG AAT CTG ATC 384 GGC CAT AGC CAG GGC GGC CTG ACA TCG CGT TAT GTT GCG TCC GTT GCA 432 CCG GAA CTG GTC GCA TCG ATG ACG AGT ACG CCG CAC TGG GGC 480 TCG CAA TTC GCG GAC TTC GTC CAG CAA CTG TTG CAG ACG GAC CCG ACC 528 GGC CTG TCG TCG ACC GTG CTC GGC GCA TTC GCG AAT GCG CTC GGC ACG 576 TTG ACG A GC AGC AAC TTC AAT ACG AAC CAG AAT GCG ATT CAG GCG TTG 624 TCG GTG CTG ACG ACG GCA AAG GCC GCC GCA TAC AAC CAG AAA TTC CCG 672 AGC GCC GGT CTC GGT GCG CCG GGC TCG TGT CAA ACC GGC GCG CCA ACG 720 GAG ACT GTC GGC GGC AAT ACG CAT CTG CTT TAT TCG TGG GGC GGC ACG 768 GCG ATC CAG CCG ACA GCG ACG GTG GCC GGC GTG ACA GGG GCC GTC GAT 816 ACG AGC GTG AGC GGG GTC ACC GAT CCG GCG AAC GCG CTC GAT CCCATC ACG CTG GCA CTC CTC GGC AGC GGC ACG GTG ATG ATC AAT CGC AGC GCC 912 GGT CCG AAC GAT GGC GTC GTG TCG CAA TGC AGC GCG CGG TTT GGC CAG 960 GTG CTC GGC ACG TAT CAC TGG AAT CAC ACC GAT GCG ATC AAC CATC 1008 CTC GGC GTG CTC GGC GCG AAT GTG GAG GAT CCG GTT GCG GTA ATC CGC 1056 ACG GAC GCG AAC CGG TTG AAG CTC GCA GGC GTG 1089

 ──────────────────────────────────────────────────続 き Continuing from the front page (72) Inventor Ken Kenta 4-2-1 Takashi Takarazuka-shi, Hyogo Prefecture Sumitomo Chemical Industry Co., Ltd.

Claims (3)

[Claims] 1. A compound represented by the general formula: (Wherein, R ₁ and R ₂ are the same or different, represent a seven carbon atoms which may be substituted with one or more hydroxyl groups to 31 aliphatic hydrocarbon group, R ₃ and R ₄ is the same or different and represents an acyl group having 2 to 7 carbon atoms.) The ability to asymmetrically hydrolyze the ester bonding site of the (2R, 3S) form of N-acyl sphingoid ester represented by Is reacted with the N-acylsphingoid ester represented by the general formula 1 and subjected to asymmetric hydrolysis, and then the unreacted N-acylsphingoid ester (2S, 3R ) Is recovered and the ester is hydrolyzed to give a compound of the general formula 2 (Wherein, R ₁ and R ₂ are the same or different, represent one or more substituted with a hydroxyl group 31 from a good carbon atoms 7 also aliphatic hydrocarbon group, R ₅ and R ₆ is an optically active alcohol compound characterized in that at least one of them represents a hydrogen atom and the other represents a hydrogen atom or an acyl group having 2 to 7 carbon atoms. Manufacturing method. 2. The method for producing an optically active alcohol compound according to claim 1, wherein the N-acyl sphingoid ester represented by the general formula 1 is an erythro compound. 3. The ester hydrolase is a protein having an amino acid sequence represented by SEQ ID NO: 1 or a sequence in which one or more amino acids in the amino acid sequence are added, deleted or substituted. A method for producing the optically active alcohol compound according to claim 1 or 2.