Nucleotide sequences which code for the metY gene

Abstract

The present invention relates to isolated polynucleotides which encode proteins with O-acetylhomoserine sulfhydrylase, vectors and cells containing the polynucleotides as well as methods of preparing L-amino acids.

1 - 40 . (Cancelled).  
     
     
         41 . An isolated polynucleotide comprising: 
 (a) a polynucleotide which encodes a polypeptide comprising the amino acid sequence of SEQ ID NO:2; or    (b) a polynucleotide which hybridizes under stringent conditions to the complement of SEQ ID NO:1 and which encodes a protein with O-acetylhomoserine sulfhydrylase activity, wherein said stringent conditions comprises washing in 0.1×SSC at a temperature of from 50 to 68° C.    
     
     
         42 . The isolated polynucleotide of  claim 41 , which is (a).  
     
     
         43 . The isolated polynucleotide of  claim 41 , which is (b).  
     
     
         44 . A vector comprising the isolated polynucleotide of  claim 42 .  
     
     
         45 . A vector comprising the isolated polynucleotide of  claim 43 .  
     
     
         46 . A host cell comprising the isolated polynucleotide of  claim 42 .  
     
     
         47 . The host cell of  claim 46 , which is a coryneform bacteria.  
     
     
         48 . The host cell of  claim 46 , wherein the activity of the polypeptide encoded by the isolated polynucleotide is increased relative to the host cell without the isolated polynucleotide.  
     
     
         49 . The host cell of  claim 48 , wherein the activity of the polypeptide is increased at least 10% relative to the host cell without the isolated polynucleotide.  
     
     
         50 . A host cell comprising the isolated polynucleotide of  claim 44 .  
     
     
         51 . The host cell of  claim 50 , which is a coryneform bacteria.  
     
     
         52 . The host cell of  claim 50 , wherein the activity of the polypeptide encoded by the isolated polynucleotide is increased relative to the host cell without the isolated polynucleotide.  
     
     
         53 . The host cell of  claim 52 , wherein the activity of the polypeptide is increased at least 10% relative to the host cell without the isolated polynucleotide.  
     
     
         54 . A process for preparing L-amino acids, comprising 
 culturing the host cell of  claim 46  for a time and under conditions suitable for the production of the L-amino acid; and    isolating the L-amino acid produced.    
     
     
         55 . The process of  claim 54 , wherein the L-amino acid is L-lysine and/or L-methionine.  
     
     
         56 . The process of  claim 54 , wherein the host cell comprises one or more overexpressed polynucleotides which encode a protein selected from the group consisting of 
 glycerolaldehyde 3 phosphate dehydrogenase, triose phosphate isomerase, 3-phosphoglycerate kinase, pyruvate carboxylase, aspartate kinase, cystathionine-gamma-synthase, cystathionine-gamma-lyase, and serine hydroxymethyltransferase.    
     
     
         57 . The process of  claim 54 , wherein the host cell comprises one or more attenuated genes which encode proteins selected from the group consisting of phosphoenol pyruvate carboxykinase, glucose 6-phosphate isomerase, pyruvate oxidase, homoserine kinase, threonine dehydratase, threonine synthase, meso-diaminopimelate D-dehydrogenase, phosphoenol pyruvate carboxykinase, glucose 6-phosphate isomerase, and pyruvate oxidase.  
     
     
         58 . A process for preparing L-amino acids, comprising 
 culturing the host cell of  claim 50  for a time and under conditions suitable for the production of the L-amino acid; and    isolating the L-amino acid produced.    
     
     
         59 . The process of  claim 58 , wherein the L-amino acid is L-lysine and/or L-methionine.  
     
     
         60 . The process of  claim 58 , wherein the host cell comprises one or more overexpressed polynucleotides which encode a protein selected from the group consisting of 
 glycerolaldehyde 3 phosphate dehydrogenase, triose phosphate isomerase, 3-phosphoglycerate kinase, pyruvate carboxylase, aspartate kinase, cystathionine-gamma-synthase, cystathionine-gamma-lyase, and serine hydroxymethyltransferase.    
     
     
         61 . The process of  claim 58 , wherein the host cell comprises one or more attenuated genes which encode proteins selected from the group consisting of phosphoenol pyruvate carboxykinase, glucose 6-phosphate isomerase, pyruvate oxidase, homoserine kinase, threonine dehydratase, threonine synthase, meso-diaminopimelate D-dehydrogenase, phosphoenol pyruvate carboxykinase, glucose 6-phosphate isomerase, and pyruvate oxidase.  
     
     
         62 . A method of preparing an L-amino acid containing feedstuff additive, comprising 
 (a) culturing the host cell of  claim 46  in a fermentation broth for a time and under conditions suitable for the production of the L-amino acid;    (b) concentrating the L-amino acid produced;    (c) removing an amount of 0 to 100 wt % of biomass formed during the culturing; and    (d) drying the fermentation broth obtained in one or both of (b) and (c) to obtain the animal feedstuff additive.    
     
     
         63 . A method of preparing an L-amino acid containing feedstuff additive, comprising 
 (a) culturing the host cell of  claim 50  in a fermentation broth for a time and under conditions suitable for the production of the L-amino acid;    (b) concentrating the L-amino acid produced;    (c) removing an amount of 0 to 100 wt % of biomass formed during the culturing; and    (d) drying the fermentation broth obtained in one or both of (b) and (c) to obtain the animal feedstuff additive.