Production of amino acids and enzymes used therefor

Abstract

A process for producing α-amino acid from starting material comprising α-amino amide enantiomers (A) and (B) is such that enantiomer (A) is converted preferentially over enantiomer (B) so that a time independent excess of at least 90%, preferably at least 98% is given. The reaction is catalysed by an amidase which may in particular be produced by specific Rhodococcus species.

1 . A process of converting an α-amino amide to an α-amino acid comprising conducting a conversion reaction catalysed by an amidase enzyme, in which the α-amino amide starting material comprises amide enantiomers (A) and (B) and in the conversion reaction enantiomer (A) is converted preferentially over enantiomer (B), 
 characterised in that the amidase enzyme is produced by Rhodococcus NCIMB 40795 or  Rhodococcus wratslaviensis  NCIMB 13082.  
 
     
     
         2 . A process according to  claim 1  in which the amidase enzyme is capable of converting enantiomer (A) such that it gives an enantiomeric excess of at least 98% independently of the conversion time.  
     
     
         3 . A process according to either preceding claim in which the α-amino amide starting material is a racemic mixture of enantiomers (A) and (B).  
     
     
         4 . A process according to  claim 1  or  claim 2  in which the enantiomer (B) is present in excess over enantiomer (A) for at least 30 minutes during the conversion reaction.  
     
     
         5 . A process according to  claim 4  in which the amount of enantiomer (B) is at least 150%, preferably at least 200%, of the amount of enantiomer (A) for at least 30 minutes during the conversion reaction.  
     
     
         6 . A process according to any preceding claim in which the α-amino amide has the formula I:  
       
         
           
           
               
               
           
         
       
       in which R is alkyl, cycloalkyl, alkenyl, cycloalkenyl, aryl, alkaryl, aralkyl, R 1 NHCOR 1 , R 1 CONHR 1 , SO 2 R 1  or SO 2 NHR 1  in which R 1  is alkyl, cycloalkyl, alkenyl, cycloalkenyl, aryl, alkaryl or aralkyl, or substituted versions of any of these, preferably C 4  to C 9  linear or branched alkyl or alkenyl, cyclic alkyl or alkenyl, phenyl, or substituted phenyl in which the substituent is selected from para-CH 3 , meta-CH 3 , ortho-CH 3 , para-CF 3 , para-Et, para-(CH 3 ) 3 C, para-Cl, para-CH 3 (CH 2 ) 3 O and para-OH.  
     
     
         7 . A process according to any preceding claim in which the α-amino acid is an unnatural amino acid.  
     
     
         8 . A process according to any preceding claim in which the enantiomer (A) is the L-enantiomer and is converted into the L α-amino acid.  
     
     
         9 . A process according to any preceding claim in which an enantiomeric excess of at least 98% is given independently of the conversion time.  
     
     
         10 . A process according to any preceding claim which is a fed batch process.  
     
     
         11 . A process according to any preceding claim in which the conversion reaction has a duration of at least 12 hours, preferably at least 24 hours.  
     
     
         12 . A process according to any preceding claim in which the α-amino acid corresponding to enantiomer (A) is present in the reaction mixture in excess over enantiomer (A) for at least 30 minutes during the conversion reaction.  
     
     
         13 . A process according to  claim 14  in which the ratio of α-amino acid corresponding to enantiomer (A) to enantiomer (A) itself is at least 3:2, preferably at least 2:1.  
     
     
         14 . A process of converting an α-amino amide to an α-amino acid comprising conducting a conversion reaction in which the α-amino amide starting material comprises amide enantiomers (A) and (B) and in the conversion reaction enantiomer (A) is converted preferentially over enantiomer (B), 
 characterised in that the conversion is brought about by contacting the starting material with cell material from Rhodococcus NCIMB 40795 or  Rhodococcus wratslaviensis  NCIMB 13082.  
 
     
     
         15 . A process of converting an α-methyl amide or an α-aminomethyl amide to an α-methyl acid or an α-aminomethyl acid, comprising conducting a conversion reaction catalysed by an amidase enzyme, in which the α-methylamide or α-aminomethyl amide starting material comprises amide enantiomers (A) and (B) and in the conversion reaction enantiomer (A) is converted preferentially over enantiomer (B), characterised in that the amidase enzyme is produced by Rhodococcus NCIMB 40795 or  Rhodococcus wratslaviensis  NCIMB 13082.  
     
     
         16 . A process of converting an α-methyl amide or an α-aminomethyl amide to an α-methyl acid or an α-aminomethyl acid, comprising conducting a conversion reaction catalysed by an amidase enzyme, in which the α-methylamide or α-aminomethyl amide starting material comprises amide enantiomers (A) and (B) and in the conversion reaction enantiomer (A) is converted preferentially over enantiomer (B), 
 characterised in that the amidase enzyme is capable of converting enantiomer (A) such that it gives an enantiomeric excess of at least 90% independently of the conversion time  
 characterised in that the conversion is brought about by contacting the starting material with cell material from Rhodococcus NCIMB 40795 or  Rhodococcus wratslaviensis  NCIMB 13082.  
 
     
     
         17 . Use of a product of a process of any of  claims 1  to  16  as an intermediate for the production of a pharmaceutical or agrochemical.  
     
     
         18 . A process for the production of a pharmaceutical or agrochemical comprising using as an intermediate a product of a process of any of  claims 1  to  16 .  
     
     
         19 . A microorganism which is Rhodococcus NCIMB 40795 or a mutant thereof having the ability to produce an amidase.  
     
     
         20 . An amidase enzyme obtainable by culturing a microorganism according to claim  19 .