Process for the preparation of (s)-2-amino-non-8-enoic acid

Abstract

Disclosed herein is a process for preparing enantioenriched (S)-2-aminonon-8-enoic acid by amination of 2-oxonon-8-enoic acid in the presence of an enzyme and an ammonia source.

1 . A method for preparing enantioenriched 2-aminonon-8-enoic acid, comprising aminating 2-oxonon-8-enoic acid in the presence of an ammonia source and a leucine dehydrogenase (LeuDH) from  Chlamydia pneumonia, Thermoactinomyces intermedius, Bacillus subtilis, Bacillus licheniformis, Geobacillus stearothermophilus , or  Bacillus sphaericus.    
     
     
         2 . (canceled) 
     
     
         3 . (canceled) 
     
     
         4 . The method of  claim 1 , wherein the LeuDH comprises a leucine dehydrogenase mutant. 
     
     
         5 . The method of  claim 1 , wherein the aminating is conducted in the presence of nicotinamide adenine dinucleotide (NAD+), D-glucose, and a glucose dehydrogenase. 
     
     
         6 . The method of  claim 1 , wherein the ammonia source comprises an ammonium chloride and ammonium hydroxide buffer solution. 
     
     
         7 . The method of  claim 1 , further comprising the step of preparing a compound of formula (IV): 
       
         
           
           
               
               
           
         
         wherein R 1  is lower alkyl; 
         comprising reacting a reagent of formula (II) with a compound of formula (III), 
       
       
         
           
           
               
               
           
         
         wherein independently for each occurrence 
         R 1  is lower alkyl; and M is selected from Li, MgCl, MgBr, and MgI. 
       
     
     
         8 . The method of  claim 7 , wherein R 1  is ethyl. 
     
     
         9 . The method of  claim 7 , wherein M is MgBr. 
     
     
         10 . The method of  claim 7 , wherein R 1  is ethyl; and M is MgBr. 
     
     
         11 . The method of  claim 7 , further comprising hydrolyzing the compound of formula (IV) to produce 2-oxonon-8-enoic acid. 
     
     
         12 . (canceled) 
     
     
         13 . The method of  claim 1 , wherein the LeuDH is from  Chlamydia pneumonia.    
     
     
         14 . The method of  claim 1 , wherein the LeuDH is from  Thermoactinomyces intermedius.    
     
     
         15 . The method of  claim 1 , wherein the LeuDH is from  Bacillus subtilis.    
     
     
         16 . The method of  claim 1 , wherein the LeuDH is from  Bacillus licheniformis.    
     
     
         17 . The method of  claim 1 , wherein the LeuDH is from  Geobacillus stearothermophilus.    
     
     
         18 . The method of  claim 1 , wherein the LeuDH is from  Bacillus sphaericus.    
     
     
         19 . The method of  claim 1 , wherein the LeuDH is a wild-type enzyme, and comprises an amino acid sequence GPAXGG (SEQ ID NO: 3); wherein X represents leucine. 
     
     
         20 . The method of  claim 1 , wherein the LeuDH is a mutant enzyme, and comprises an amino acid sequence GPAXGG (SEQ ID NO: 3); wherein X represents an amino acid residue that is not leucine. 
     
     
         21 . The method of  claim 20 , wherein X represents glycine, valine, isoleucine, alanine, serine, or threonine.