JP2003093089A - Method for producing (s)-3-quinuclidinol - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a method for producing (S)-3-quinuclidinol useful as an intermediate of physiologically active substances (e.g. medicaments). SOLUTION: This method for producing the (S)-3-quinuclidinol comprises making an enzyme (a) or (b) act on 3-quinuclidinone or a salt thereof, wherein the enzyme (a) is an enzyme having a specific amino acid sequence, and the enzyme (b) is an enzyme having an amino acid sequence where one or several amino acid(s) are deleted, substituted or added from/for/to the amino acid sequence in the enzyme (a) and also having the enzymatic activity of reducing 3-quinuclidinone into the objective (S)-3-quinuclidinol.

Description

Description: BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a method for producing (S) -3-quinuclidinol, and more particularly, to the action of a certain enzyme on 3-quinuclidinone or a salt thereof. The present invention relates to a method for producing (S) -3-quinuclidinol, which is a feature. 2. Description of the Related Art Numerous physiologically active substances having a quinuclidine ring in a molecule, such as quinidine and quinine, are known. (S) -3-quinuclidinol is a compound useful as an intermediate of these physiologically active substances (for example, drugs), and development of an industrially advantageous production method thereof is desired. [0003] The present inventors have proposed (S) -3.
-As a result of intensive studies on the production method of quinuclidinol, 3-
A quinuclidinone or a salt thereof is an enzyme having an amino acid sequence represented by SEQ ID NO: 1 or an amino acid sequence in which one or several amino acids are deleted, substituted or added in SEQ ID NO: 1, and 3-quinuclidinone is represented by (S) The present inventors have found that (S) -3-quinuclidinol can be obtained by allowing an enzyme having an enzymatic activity to reduce -3-quinuclidinol to act, thereby completing the present invention. That is, the present invention provides the following inventions. Formula (1) 3-quinuclidinone or a salt thereof represented by the following a)
Or b) is acted upon to obtain formula (2) (S) -3-quinuclidinol represented by the following formula (hereinafter, referred to as the production method of the present invention). a) An enzyme having the amino acid sequence represented by SEQ ID NO: 1. b) An enzyme having an amino acid sequence in which one or several amino acids are deleted, substituted or added in SEQ ID NO: 1 and having an enzymatic activity of reducing 3-quinuclidinone to (S) -3-quinuclidinol. (Hereinafter a) and b) are collectively referred to as the present enzyme. BEST MODE FOR CARRYING OUT THE INVENTION A gene sequence encoding an enzyme having the amino acid sequence represented by SEQ ID NO: 1 which can be used in the production method of the present invention is represented by SEQ ID NO: 2 (Appl. Mi)
crobiol. Biotechnol (1999) 52,386-392). The gene having the base sequence encoding the amino acid sequence of the enzyme may be a naturally occurring gene obtained by subjecting a naturally occurring gene to a mutation treatment (partial mutation introduction method, mutation treatment, etc.). You may. The present enzyme can be produced, for example, by culturing a microorganism containing the gene represented by SEQ ID NO: 2 and producing an enzyme having the amino acid sequence represented by SEQ ID NO: 1 when the gene is expressed. . A microorganism that contains the gene represented by SEQ ID NO: 2 and that produces an enzyme having the amino acid sequence represented by SEQ ID NO: 1 when the gene is expressed is a naturally occurring microorganism,
It may be a transformed microorganism into which a gene having the nucleotide sequence of SEQ ID NO: 2 has been introduced. Here, a method for preparing a transformed microorganism into which a gene having the nucleotide sequence of SEQ ID NO: 2 has been introduced will be described. As a host cell into which a gene having the nucleotide sequence represented by SEQ ID NO: 2 is introduced, for example, Escherichi
a, Bacillus, Corynebacterium, Staphylococcus, Stre
ptomyces, Saccharomyces, Kluyveromyces and Aspergil
Microorganisms belonging to the genus lus can be mentioned. Method for introducing the gene into a host cell is not particularly limited as long as the normal methods used depending on the cell as a host, for example, "Molecular Cloning: ALaboratory Manual 2 ^nd ed
ition "(1989), Cold Spring Harbor Laboratory Pres
s, `` Current Protocols in Molecular Biology '' (198
7), John Wiley & Sons, Inc. ISBNO-471-50338-X, etc.
oration: Gene Pulser /E.coli Pulser System '' Bio-Ra
d Laboratories, (1993) and the like. [0008] The transformed microorganism into which the gene has been introduced is
Selection can be performed using the phenotype of the selectable marker gene contained in the vector used when introducing the gene into the host cell as an index. The fact that the transformed microorganism has the gene means that the transformed microorganism
Is prepared, and the prepared DNA is subjected to a conventional method (confirmation of restriction enzyme site, nucleotide sequence, etc.) described in, for example, "Molecular cloning" (J. Sambrook et al., Cold Spring Harbor, 1989). Analysis, Southern hybridization, etc.). Next, to produce the enzyme used in the production method of the present invention, it contains the gene represented by SEQ ID NO: 2,
A method for culturing a microorganism that produces an enzyme having the amino acid sequence represented by SEQ ID NO: 1 when the gene is expressed will be described. [0010] As a medium for culturing the microorganism, there can be used various kinds of media which contain carbon sources and nitrogen sources, organic salts, inorganic salts and the like which are usually used for culturing microorganisms. Examples of the carbon source include sugars such as glucose, dextrin and sucrose, sugar alcohols such as glycerol, organic acids such as fumaric acid, citric acid and pyruvic acid, animal and vegetable oils, and molasses. The amount of these carbon sources added to the medium is usually 0.1 to 20% based on the total amount of the medium.
(W / v). Nitrogen sources include meat extract, peptone, yeast extract, malt extract, soy flour, corn steep,
Natural organic nitrogen sources and amino acids such as liquor (Corn Steep Liquor), cottonseed flour, dried yeast and casamino acid, ammonium salts and nitrates of inorganic acids such as sodium nitrate, ammonium chloride, ammonium sulfate and ammonium phosphate, and ammonium fumarate And ammonium salts of organic acids such as ammonium citrate, and organic or inorganic nitrogen sources such as urea. Of these, ammonium salts of organic acids, natural organic nitrogen sources, amino acids and the like can often be used as carbon sources. The amount of addition of the nitrogen source may be usually about 0.1 to 30% (w / v) based on the total amount of the medium. Examples of the organic and inorganic salts include chlorides, sulfates, acetates, carbonates and phosphates of potassium, sodium, magnesium, iron, manganese, cobalt, zinc and the like. Is sodium chloride,
Examples thereof include potassium chloride, magnesium sulfate, ferrous sulfate, manganese sulfate, cobalt chloride, zinc sulfate, copper sulfate, sodium acetate, calcium carbonate, sodium carbonate, monopotassium hydrogen phosphate, and dipotassium hydrogen phosphate. The amount of organic or inorganic salt added is usually
It is good to be about 0.0001-5% (w / v). Furthermore, a host cell into which a gene obtained by operably connecting a promoter of the type induced by allolactose, such as a tac promoter, a trc promoter, a lac promoter, and a gene encoding the present enzyme, is introduced. In some cases, as an inducer for inducing the production of the enzyme, for example, isopropyl thio-β-D-galactoside (IPT
G) may be added in a small amount to the medium. The culturing can be carried out according to a method usually used for culturing microorganisms.
Reciprocating shaking culture, Jar Fermente
r) Methods such as liquid culture such as culture and tank culture, and solid culture are possible. When using a jar fermenter,
It is necessary to introduce sterile air into the jar fermenter, and usually aeration conditions of about 0.1 to about 2 times / min of the volume of the culture solution are used. The culture temperature is preferably in the range of 35 to 42 ° C, and the pH of the medium is preferably in the range of about 6 to about 8. The culturing time varies depending on the culturing conditions, but is usually about 1 hour.
Days to about 5 days. In the production method of the present invention, for example, cells containing the present enzyme obtained as described above, processed cells thereof, or purified products of the present enzyme can be used. Here, the treated cells may be, for example, freeze-dried cells, organic solvent-treated cells, dried cells, crushed cells,
Autolysed cells of the cells, sonicated cells, cell extracts,
Alkali-treated cells can be mentioned, and those immobilized by these commonly used methods can also be mentioned. The purified product of the present enzyme can be produced, for example, by purifying the present enzyme from a culture of a microorganism having the present enzyme. As a method for purifying the present enzyme from a culture of a microorganism having the present enzyme, a method used in ordinary protein purification can be applied, and examples thereof include the following methods. First, cells are collected from the culture of the microorganism by centrifugation or the like, and then subjected to ultrasonic treatment, dynomill treatment, a physical crushing method such as French press treatment, or a cell lysing enzyme such as a surfactant or lysozyme. Crushing is performed according to the chemical crushing method used. Insoluble matter is removed from the obtained crushed liquid by centrifugation, filtration with a membrane filter, etc. to prepare a cell-free extract, which is then subjected to cation exchange chromatography, anion exchange chromatography, hydrophobic chromatography, gel chromatography, etc. The present reductase can be purified by fractionation using the separation and purification method described above as appropriate. Examples of the carrier used for chromatography include resin carriers such as cellulose, dextran, and agarose, into which a carboxymethyl (CM) group, a DEAE group, a phenyl group, a butyl group, or the like has been introduced. A commercially available carrier-packed column can also be used, for example,
Q-Sepharose FF, Phenyl-Sepharose HP (trade names, all manufactured by Amersham Pharmacia Biotech), T
SK-gel G3000SW (trade name, manufactured by Tosoh Corporation)
And the like. Next, the production method of the present invention will be described.
In the production method of the present invention, the reaction for converting 3-quinuclidinone or a salt thereof to (S) -3-quinuclidinol is 3-
This is achieved by allowing the present enzyme to act on quinuclidinone. Examples of the salt of 3-quinuclidine used in the reaction include salts of inorganic acids such as hydrochloride and salts of organic acids such as acetate. The reaction is usually performed in the presence of water, and water may be in the form of a buffer. In this case, examples of the buffer include alkali metal phosphates such as sodium phosphate and potassium phosphate. And alkali metal salts of acetic acid such as sodium acetate and potassium acetate. In this case, the pH can be appropriately changed within a range in which the reaction proceeds. When a buffer is used as a solvent, the amount is usually 100 parts by weight or less based on 1 part by weight of 3-quinuclidinone or a salt thereof. The reaction temperature is from 0 to 70 ° C, preferably from 10 to 40 ° C, in view of the stability and reaction rate of the enzyme. The reaction can be carried out in the presence of an organic solvent in addition to water. As the organic solvent in this case, for example, tetrahydrofuran, t-butyl methyl ether,
Ethers such as isopropyl ether, hydrocarbons such as toluene, hexane, cyclohexane, heptane, isooctane and decane, t-butanol, methanol,
Examples thereof include alcohols such as ethanol, isopropanol and n-butanol, sulfoxides such as dimethyl sulfoxide, ketones such as acetone, nitriles such as acetonitrile, and mixtures thereof. The amount of the organic solvent used in the reaction is usually 100 times by weight or less, preferably 70 times by weight or less, based on 3-quinuclidinone or a salt thereof. The reaction further comprises a coenzyme (eg, NAD
H, NADPH). The amount of coenzyme used in the reaction is usually 0.5 times by weight or less, preferably 0.1 times by weight or less, based on 3-quinuclidinone or a salt thereof. When a coenzyme is added to the reaction, it is preferable to further add the following in order to increase the efficiency of the coenzyme. 1) Compounds such as formic acid, glucose, isopropanol, 2-butanol, 2-pentanol, 2-hexanol, 2-heptanol and 2-octanol The amount of these compounds used in this case is 3-quinuclidinone or It is 100 times by weight or less, preferably 10 times by weight or less based on the salt. 2) Dehydrogenases such as formate dehydrogenase and glucose dehydrogenase The amount of dehydrogenase used in this case is 0.1 weight times or less, preferably 0.05 weight times, based on 3-quinuclidinone or a salt thereof. Less than twice. The reaction can be carried out, for example, by mixing water, 3-quinuclidinone or a salt thereof, the present enzyme and, if necessary, a coenzyme, an organic solvent and the like, and stirring and shaking. The end point of the reaction can be determined, for example, by following the amount of the starting compound in the reaction solution by liquid chromatography, gas chromatography or the like. The range of the reaction time is usually from 5 minutes to 4 days. After completion of the reaction, for example, the reaction mixture is extracted with an organic solvent such as hexane, heptane, tert-butyl methyl ether, ethyl acetate, toluene, and the like, and the organic layer is dried and concentrated to isolate the desired product. Can be released. The target substance can be purified by column chromatography or the like, if necessary. In the production method of the present invention, a) an amino acid sequence having the amino acid sequence shown in SEQ ID NO: 1 instead of b) an amino acid sequence in which one or several amino acids are deleted, substituted or added in SEQ ID NO. And an enzyme having an enzymatic activity of reducing 3-quinuclidinone to (S) -3-quinuclidinol. In this case, for example, a well-known technique for causing a point mutation or the like in the DNA shown in SEQ ID NO: 2, for example, in the DNA,
A site-directed mutagenesis method; a method of selectively cleaving DNA and then removing or adding selected nucleotides to ligate the DNA; or a method in which SEQ ID NO: 1 or It consists of an amino acid sequence in which several amino acids have been deleted, substituted or added, and has 3-quinuclidinone as (S)
A gene encoding an enzyme having an enzyme activity of reducing to -3-quinuclidinol is prepared, and a transformant is prepared, cultured, reacted, and the like in the same manner as described above to carry out the production method of the present invention. EXAMPLES The present invention will be described in more detail with reference to production examples and the like, but the present invention is not limited to these examples. Production Example A liquid medium (tryptone 10 in 1000 ml of water) was placed in a flask.
g, 5 g of yeast extract and 5 g of sodium chloride were dissolved, and pH was adjusted to 7.0 by dropwise addition of a 1N aqueous sodium hydroxide solution. ) Add 900ml and sterilize,
100 μg / ml ampicillin, isopropyl thio-β-
D-galactoside (IPTG) was added to 0.4 mM, and a plasmid pUAR containing the DNA shown in SEQ ID NO: 2 (Accession number: FERM P-1812) was added thereto.
The transformant E. coli JM109 / pUAR obtained by transforming the E. coli JM109 strain by the conventional method in 7) was inoculated with 1 ml of a culture solution cultured in a liquid medium having the above composition, and cultured with shaking at 37 ° C. for 14 hours. The culture was centrifuged (15000 × g, 15
For 30 minutes at 4 ° C.), and the obtained cells are treated with a 50 mM monopotassium phosphate-dipotassium phosphate buffer (pH 7.0) 30 m
l, and the suspension is centrifuged (15000 xg,
(15 minutes, 4 ° C.) to obtain washed cells. NAD was added to 7.5 ml of a 50 mM monopotassium phosphate-dipotassium phosphate buffer (pH 7.0) containing 5% of 2-propanol.
⁺ 7.5 mg and adding the washed cells 2.4 g. To this, 79.5 mg (0.50 mmol) of 3-quinuclidinone hydrochloride was added, and the mixture was stirred at room temperature for 4 days. Thereafter, 3 ml of water was poured into the reaction solution, stirred, and then centrifuged. 5 g of sodium carbonate was added to the resulting supernatant, and the mixture was vigorously shaken for 1 minute.
After vigorous shaking for minutes, it was centrifuged. The obtained organic layer and aqueous layer were separated, 7.5 ml of ethyl acetate was added to the aqueous layer, and the mixture was vigorously shaken for 1 minute. The operation of liquid separation was repeated 5 times, and the obtained organic layers were combined and concentrated. And 10 mg of (S) -3-quinuclidinol. (Chemical purity 76%, optical purity 100% ee) MS (EI, m / z) 127 (M ⁺ ), 112, 9
8, 42 The absolute configuration of the obtained (S) -3-quinuclidinol was determined by the obtained compound, commercially available (R)
-3-Quinuclidinol and racemic 3-quinuclidinol were subjected to gas chromatography analysis using an optically active column, and the results were determined. Chemical Purity Analysis Conditions Gas chromatography column: DB-1 (Shinwa processing) 0.53 mmφ × 30
m Membrane pressure 1.5 μm Inlet temperature: 120 ° C. Column chamber temperature: 50 ° C. (4 ° C./min)→170° C. Detector temperature: 300 ° C. Carrier gas: helium 10 ml / min Optical purity analysis gas chromatography Column: Beta DEX120 0.25mmφ × 30m
Membrane pressure 0.25 μm Inlet temperature: 250 ° C. Column chamber temperature: 140 ° C. Detector (FID) temperature: 250 ° C. Carrier gas: helium 1 ml / min Split ratio 1
/ 50 Retention time: S-form = 18.5 minutes, R-form = 19.0 minutes According to the present invention, a (S) -3-quinuclidinol compound useful as an intermediate for producing a physiologically active substance Can be manufactured. [Sequence List] SEQUENCE LISTING <110> Sumitomo Chemical Co., Ltd. <120> Preparation of (S) -3-Quinuclidinol <130> P153361 <160> 2 <210> 1 <211> 385 <212> PRT <213> Corynebacterium sp. <400> 1 Met Lys Ala Ile Gln Tyr Thr Arg Ile Gly Ala Glu Pro Glu Leu Thr 1 5 10 15 Glu Ile Pro Lys Pro Glu Pro Gly Pro Gly Glu Val Leu Leu Glu Val 20 25 30 Thr Ala Ala Gly Val Cys His Ser Asp Asp Phe Ile Met Ser Leu Pro 35 40 45 Glu Glu Gln Tyr Thr Tyr Gly Leu Pro Leu Thr Leu Gly His Glu Gly 50 55 60 Ala Gly Lys Val Ala Ala Val Gly Glu Gly Val Glu Gly Leu Asp Ile 65 70 75 80 Gly Thr Asn Val Val Val Tyr Gly Pro Trp Gly Cys Gly Asn Cys Trp 85 90 95 His Cys Ser Gln Gly Leu Glu Asn Tyr Cys Ser Arg Ala Gln Glu Leu 100 105 110 Gly Ile Asn Pro Pro Gly Leu Gly Ala Pro Gly Ala Leu Ala Glu Phe 115 120 125 Met Ile Val Asp Ser Pro Arg His Leu Val Pro Ile Gly Asp Leu Asp 130 135 140 Pro Val Lys Thr Val Pro Leu Thr Asp Ala Gly Leu Thr Pro Tyr His 145 150 155 160 Ala Ile Lys Arg Ser Leu Pro Lys Leu Arg Gly Gly Ser Tyr Ala Val 165 170 175 Val Ile Gly Thr Gly Gly Leu Gly His Val Ala Ile Gln Leu Leu Arg 180 185 190 His Leu Ser Ala Ala Thr Val Ile Ala Leu Asp Val Ser Ala Asp Lys 195 200 205 Leu Glu Leu Ala Thr Lys Val Gly Ala His Glu Val Val Leu Ser Asp 210 215 220 Lys Asp Ala Ala Glu Asn Val Arg Lys Ile Thr Gly Ser Gln Gly Ala 225 230 235 240 Ala Leu Val Leu Asp Phe Val Gly Tyr Gln Pro Thr Ile Asp Thr Ala 245 250 255 Met Ala Val Ala Gly Val Gly Ser Asp Val Thr Ile Val Gly Ile Gly 260 265 270 Asp Gly Gln Ala His Ala Lys Val Gly Phe Phe Gln Ser Pro Tyr Glu 275 280 285 Ala Ser Val Thr Val Pro Tyr Trp Gly Ala Arg Asn Glu Leu Ile Glu 290 295 300 Leu Ile Asp Leu Ala His Ala Gly Ile Phe Asp Ile Gly Gly Gly Asp 305 310 315 320 Leu Gln Ser Arg Gln Arg Cys Arg Ser Val Ser Thr Thr Gly Cys Arg 325 330 335 Asn Ala Gln Arg Pro Cys Gly Cys Gly Pro Trp Ser Val Val Pro Thr 340 345 350 Ala Val Glu Arg Gln Arg Lys Asn Thr Asp Ala Arg Pro Asn Ser Ile 355 360 365 Arg Pro Gly Ile Ser Val Arg Asn Ser Val Cys Ala Ser Cys Thr Pro 370 375 380 Arg 385 <210> 2 <211> 1158 <212> DNA <213> Corynebacterium sp. <220><221> CDS <222> (1) .. (1158) <400> 2 atg aag gcg atc cag tac acg cga atc ggc gcg gaa ccc gaa ctc acg 48 Met Lys Ala Ile Gln Tyr Thr Arg Ile Gly Ala Glu Pro Glu Leu Thr 1 5 10 15 gag att ccc aaa ccc gag ccc ggt cca ggt gaagt ctg gaa gtc 96 Glu Ile Pro Lys Pro Glu Pro Gly Pro Gly Glu Val Leu Leu Glu Val 20 25 30 acc gct gct ggc gtc tgc cac tcg gac gac ttc atc atg agc ctg ccc 144 Thr Ala Ala Gly Val Cys His Ser Asp Asp Phe Ile Met Ser Leu Pro 35 40 45 gaa gag cag tac acc tac ggc ctt ccg ctc acg ctc ggc cac gaa ggc 192 Glu Glu Gln Tyr Thr Tyr Gly Leu Pro Leu Thr Leu Gly His Glu Gly 50 55 60 gca ggc aag gtc gcc gcc gtc ggc gag ggt gtc gaa ggt ctc gac atc 240 Ala Gly Lys Val Ala Ala Val Gly Glu Gly Val Glu Gly Leu Asp Ile 65 70 75 80 gga acc aat gtc gtc gtc tac ggg cct tgg ggt tgc ggc aac tgt tgg Thr Asn Val Val Val Tyr Gly Pro Trp Gly Cys Gly Asn Cys Trp 85 90 95 cac tgc tca ca a gga ctc gag aac tat tgc tct cgc gcc caa gaa ctc 336 His Cys Ser Gln Gly Leu Glu Asn Tyr Cys Ser Arg Ala Gln Glu Leu 100 105 110 gga atc aat cct ccc ggt ctc ggt gca ccc ggc gcg ttg gcc gtt Gly Ile Asn Pro Pro Gly Leu Gly Ala Pro Gly Ala Leu Ala Glu Phe 115 120 125 atg atc gtc gat tct cct cgc cac ctt gtc ccg atc ggt gac ctc gac 432 Met Ile Val Asp Ser Pro Arg His Leu Val Pro Ile Gly Asp Leu Asp 130 135 140 ccg gtc aag acg gtg ccg ctg acc gac gcc ggt ctg acg ccg tat cac 480 Pro Val Lys Thr Val Pro Leu Thr Asp Ala Gly Leu Thr Pro Tyr His 145 150 155 160 gcg atc aag cgt tct ctg ccg aaa ctt cgc gga ggc tcg tac gcg gtt 528 Ala Ile Lys Arg Ser Leu Pro Lys Leu Arg Gly Gly Ser Tyr Ala Val 165 170 175 gtc att ggt acc ggc ggt ctc ggc cac gtc gct att cag ctc ctc cgc Gly G Val I Gly Leu Gly His Val Ala Ile Gln Leu Leu Arg 180 185 190 cac ctc tcg gcg gca acg gtc atc gct ttg gac gtg agc gcg gac aag 624 His Leu Ser Ala Ala Thr Val Ile Ala Leu Asp Val Ser Ala Asp Lys 195 200 205 ct c gaa ctg gca acc aag gta ggc gct cac gaa gtg gtt ctg tcc gac 672 Leu Glu Leu Ala Thr Lys Val Gly Ala His Glu Val Val Leu Ser Asp 210 215 220 aag gac gcg gcc gag aac gtc cgc aag atc act gga agt cagt ggc gcc 720 Lys Asp Ala Ala Glu Asn Val Arg Lys Ile Thr Gly Ser Gln Gly Ala 225 230 235 240 gca ttg gtt ctc gac ttc gtc ggc tac cag ccc acc atc gac acc gcg 768 Ala Leu Val Leu Asp Phe Val Gly Tyr Gln Pro Thr Ile Asp Thr Ala 245 250 255 atg gct gtc gcc ggc gtc gga tca gac gtc acg atc gtc ggg atc ggg 816 Met Ala Val Ala Gly Val Gly Ser Asp Val Thr Ile Val Gly Ile Gly 260 265 270 270 gac ggc cag gcc cac gcc aaa gtc ggg ttc ttc caa agt cct tac gag 864 Asp Gly Gln Ala His Ala Lys Val Gly Phe Phe Gln Ser Pro Tyr Glu 275 280 285 gct tcg gtg aca gtt ccg tat tgg ggt gcc cgc aac gag ttg atcga Atta gga ttg atc ga Val Thr Val Pro Tyr Trp Gly Ala Arg Asn Glu Leu Ile Glu 290 295 300 ttg atc gac ctc gcc cac gcc ggc atc ttc gac atc ggc ggt gga gac 960 Leu Ile Asp Leu Ala His Ala Gly Ile Phe Asp Ile Gly Gly Gly Gly 30 5 310 315 320 ctt cag tct cga caa cgg tgc cga agc gta tcg acg act ggc tgc cgg 1008 Leu Gln Ser Arg Gln Arg Cys Arg Ser Val Ser Thr Thr Gly Cys Arg 325 330 335 aac gct cag cgg ccg tgc ggt tgt ggt ccc tgg tct gta gta ccg aca 1056 Asn Ala Gln Arg Pro Cys Gly Cys Gly Pro Trp Ser Val Val Pro Thr 340 345 350 gcg gta gaa cga cag cgg aaa aac act gat gcc cgg ccg aat tcg att 1104 Ala Val Glu Arg Gln Arg Lys Asn Thr Asp Ala Arg Pro Asn Ser Ile 355 360 365 cgg ccg ggc atc agt gtc aga aat tcg gtg tgc gct agc tgc acg cct 1152 Arg Pro Gly Ile Ser Val Arg Asn Ser Val Cys Ala Ser Cys Thr Pro 370 375 380 380 cga tga 1158 Arg 385

Claims (1)

[Claim 1] (S)-characterized by reacting the following a) or b) on 3-quinuclidinone or a salt thereof.
A method for producing 3-quinuclidinol. a) An enzyme having the amino acid sequence represented by SEQ ID NO: 1. b) An enzyme comprising an amino acid sequence in which one or several amino acids are deleted, substituted or added in SEQ ID NO: 1 and having an enzymatic activity of reducing 3-quinuclidinone to (S) -3-quinuclidinol.