Polynucleotides Encoding Polypeptides Involved in Amino Acid Biosynthesis in Methylophilus Methylotrophus

Abstract

The present invention provides polypeptides and polynucleotides involved in amino acid biosynthesis in Methylophilus methylotrophus and methods of producing amino acids in microorganisms having enhanced or attenuated expression of these polypeptides and/or polynucleotides.

1 . An isolated polynucleotide, which encodes a protein comprising the amino acid sequences of SEQ ID NO:30 and SEQ ID NO:32.  
   
   
       2 . A vector comprising the isolated polynucleotide of  claim 1 .  
   
   
       3 . An isolated host cell comprising the isolated polynucleotide of  claim 1 .  
   
   
       4 . The host cell of  claim 3 , which is a  Methylophilus  bacterium.  
   
   
       5 . The host cell of  claim 4 , which is a  Methylophilus methylotrophus  bacterium.  
   
   
       6 . A method of producing at least one L-amino acid comprising 
 culturing the host cell of  claim 5  for a time and under conditions suitable for producing the amino acid; and    collecting the amino acid produced.    
   
   
       7 . An isolated polynucleotide comprising the nucleotide sequences of SEQ ID NO:29 and SEQ ID NO:31.  
   
   
       8 . A vector comprising the isolated polynucleotide of  claim 7 .  
   
   
       9 . An isolated host cell comprising the isolated polynucleotide of  claim 7 .  
   
   
       10 . The host cell of  claim 9 , which is a  Methylophilus bacterium.    
   
   
       11 . The host cell of  claim 10 , which is a  Methylophilus methylotrophus  bacterium.  
   
   
       12 . A method of producing at least one L-amino acid comprising 
 culturing the host cell of  claim 11  for a time and under conditions suitable for producing the amino acid; and    collecting the amino acid produced.    
   
   
       13 . An isolated polynucleotide, which hybridizes under stringent conditions to the isolated polynucleotide of  claim 7 , wherein the stringent conditions comprise hybridization in 50% formamide, 1M NaCl, and 1% SDS at 37° C., and a wash in 0.1×SSC at 65° C., 
 and wherein said polynucleotide encodes a polypeptide having pyruvate carboxylase activity.    
   
   
       14 . A vector comprising the isolated polynucleotide of  claim 13 .  
   
   
       15 . An isolated host cell comprising the isolated polynucleotide of  claim 13 .  
   
   
       16 . The method of  claim 15 , wherein said host cell is a  Methylophilus  bacterium.  
   
   
       17 . The method of  claim 16 , wherein said host cell is a  Methylophilus methylotrophus  bacterium.  
   
   
       18 . A method of producing at least one L-amino acid comprising 
 culturing the host cell of  claim 17  for a time and under conditions suitable for producing the amino acid; and    collecting the amino acid produced.    
   
   
       19 . An isolated polynucleotide, which is at least 95% identical to the polynucleotide of  claim 7 , and wherein said polynucleotide encodes a polypeptide having pyruvate carboxylase activity.  
   
   
       20 . A vector comprising the isolated polynucleotide of  claim 19 .  
   
   
       21 . An isolated host cell comprising the isolated polynucleotide of  claim 19 .  
   
   
       22 . The host cell of  claim 21 , wherein said host cell is a  Methylophilus  bacterium.  
   
   
       23 . The host cell of  claim 22 , wherein said host cell is a  Methylophilus methylotrophus  bacterium.  
   
   
       24 . A method of producing at least one L-amino acid comprising 
 culturing the host cell of  claim 23  for a time and under conditions suitable for producing the amino acid; and    collecting the amino acid produced.