Novel amino alcohol dehydrogenase, method for producing said enzyme, and use of said enzyme

Abstract

The present invention provides amino alcohol dehydrogenases which catalyze a reaction to produce keto alcohols, ketones, aldehydes, keto acids from corresponding amino alcohols, amines, and amino acids in the presence of NAD + , and a reaction to produce amino alcohols, amines, amino acids from corresponding keto alcohols, ketones, aldehydes, and keto acids in the presence of NADH and ammonium ions. Also provided is a method for producing the enzymes and uses of the enzyme. The enzymes of the present invention can be obtained from microorganisms belonging to the genera Streptomyces, Pseudomonas, Burkholdenia, or Arthrobacter.

What is claimed is:  
     
         1 . An amino alcohol dehydrogenase that reductively converts keto alcohol into amino alcohol, and oxidatively converts amino alcohol into keto alcohol.  
     
     
         2 . The amino alcohol dehydrogenase of  claim 1 , which reductively converts keto acid into amino acid and oxidatively converts amino acid into keto acid.  
     
     
         3 . The amino alcohol dehydrogenase of  claim 1 , which reductively converts ketone or aldehyde into amine and oxidatively converts amine into ketone or aldehyde.  
     
     
         4 . The amino alcohol dehydrogenase of  claim 2 , which reductively converts ketone or aldehyde into amine and oxidatively converts amine into ketone or aldehyde.  
     
     
         5 . The amino alcohol dehydrogenase of  claim 1 , which is obtainable from a microorganism selected from the group consisting of the genera Streptomyces, Pseudomononas, Burkholdenia, and Arthrobacter.  
     
     
         6 . The amino alcohol dehydrogenase of  claim 5 , wherein the microorganism belonging to the genus Streptomyces is selected from the group consisting of the species  Streptomyces virginiae, Streptomyces griseus, Streptomyces avidinii,  and  Streptomyces pseudovenezulae.    
     
     
         7 . The amino alcohol dehydrogenase of  claim 5 , wherein the microorganism belonging to the genus Pseudomononas is the species  Pseudomonaonas fluorescens  or  Pseudomonas marginalis.    
     
     
         8 . The amino alcohol dehydrogenase of  claim 5 , wherein the microorganism belonging to the genus Burkholdenia is the species  Burkholdenia cepacia.    
     
     
         9 . The amino alcohol dehydrogenase of  claim 5 , wherein the microorganism belonging to the genus Arthrobacter is the species  Arthrobacter aurescens.    
     
     
         10 . An amino alcohol dehydrogenase having the following physicochemical properties: 
 (a) NAD(H)-dependent;    (b) a molecular weight of a part of the subunit of about 46,000 Da when determined by SDS-polyacrylamide gel electrophoresis, and of the whole molecule of about 100,000 Da when determined by gel filtration;    (c) substrate specificity, such that it acts on amino alcohols, amines, amino acids in the presence of NAD +  to produce keto alcohols, ketones, aldehydes, and keto acids, and acts on keto alcohols, ketones, aldehydes, and keto acids in the presence of NADH and ammonium ions to produce amino alcohols, amines, and amino acids;    (d) thermostability, such that it is relatively stable at 30° C. and inactivated at 40° C. or higher when heated at pH 7.0 for 30 min;    (e) optimum temperature of about 30° C. in reductive amination at pH 7.0;    (f) optimum pH of 10.0 in oxidative deamination and of 7.0 in reductive amination; and    (g) stability, such that its activity is stable in the presence of glycerol or serinol, or phenylmethylsulfonylfluoride, a protease inhibitor.    
     
     
         11 . A method for producing amino alcohol dehydrogenase, the method comprising culturing a microorganism, which produces the amino alcohol dehydrogenase of  claim 1 , and recovering the enzyme from the culture.  
     
     
         12 . A method for producing amino alcohol, the method comprising reacting keto alcohol with the amino alcohol dehydrogenase of  claim 1  in a reaction system, and recovering the corresponding amino alcohol from the reaction system.  
     
     
         13 . A method for producing amino acid, the method comprising reacting keto acid with the amino alcohol dehydrogenase of  claim 2  in a reaction system, and recovering the corresponding amino acid from the reaction system.  
     
     
         14 . A method for producing amine, the method comprising reacting ketone or aldehyde with the amino alcohol dehydrogenase of  claim 3  in a reaction system, and recovering the corresponding amine from the reaction system.  
     
     
         15 . A method for producing amine, the method comprising reacting ketone or aldehyde with the amino alcohol dehydrogenase of  claim 4  in a reaction system, and recovering the corresponding amine from the reaction system.  
     
     
         16 . A method for producing keto alcohol, the method comprising reacting amino alcohol with the amino alcohol dehydrogenase of  claim 1  in a reaction system, and recovering the corresponding keto alcohol from the reaction system.  
     
     
         17 . A method for producing keto acid, the method comprising reacting amino acid with the amino alcohol dehydrogenase of  claim 2  in a reaction system, and recovering the corresponding keto acid from the reaction system.  
     
     
         18 . A method for producing ketone or aldehyde, the method comprising reacting amine with the amino alcohol dehydrogenase of  claim 3  in a reaction system, and recovering the corresponding ketone or aldehyde from the reaction system.  
     
     
         19 . A method for producing ketone or aldehyde, the method comprising reacting amine with the amino alcohol dehydrogenase of  claim 4  in a reaction system, and recovering the corresponding ketone or aldehyde from the reaction system.  
     
     
         20 . A microorganism producing amino alcohol dehydrogenase of  claim 1 , which has the characteristics of the microorganism selected from the group consisting of  Arthrobacter aurescens  B151 identified by a deposit number of FERM P-17137,  Burkholdenia cepacia  B033 identified by a deposit number of FERM P-17138,  Pseudomonas fluorescens  B101 identified by a deposit number of FERM P-17139,  Pseudomonas marginalis  B102 identified by a deposit number of FERM P-17140,  Streptomyces griseus  TPC 33081 identified bya deposit number of FERM P-17141,  Streptomyces avidinii  A044 identified by a deposit number of FERM P-17142, and  Streptomyces pseudovenezulae  A161 identified by a deposit number of FERM P-17143.  
     
     
         21 . A method for producing amino alcohol, the method comprising reacting keto alcohol with the microorganism of  claim 20  or its treated product in a reaction system, and recovering the corresponding amino alcohol from the reaction system.  
     
     
         22 . A method for producing amino acid, the method comprising reacting keto acid with the microorganism of  claim 20  or its treated product in a reaction system, and recovering the corresponding amino acid from the reaction system.  
     
     
         23 . A method for producing amine, the method comprising reacting ketone or aldehyde with the microorganism of  claim 20  or its treated product in a reaction system, and recovering the corresponding amine from the reaction system.  
     
     
         24 . A method for producing keto alcohol, the method comprising reacting amino alcohol with the microorganism of  claim 20  or its treated product in a reaction system, and recovering the corresponding keto alcohol from the reaction system.  
     
     
         25 . A method for producing keto acid, the method comprising reacting amino acid with the microorganism of  claim 20  or its treated product in a reaction system, and recovering the corresponding keto acid from the reaction system.  
     
     
         26 . A method for producing ketone or aldehyde, the method comprising reacting amine with the microorganism of  claim 20  or its treated product in a reaction system, and recovering the corresponding ketone or aldehyde from the reaction system.