Methods of making L-amino acids in coryneform bacteria using the sigE gene

Abstract

The present invention relates to an isolated polynucleotide from Corynebacterium glutamicum comprising a polynucleotide sequence chosen from the group consisting of (a) a polynucleotide which is identical to the extent of at least 70% to a polynucleotide which codes for a polypeptide which comprises the amino acid sequence of SEQ ID NO: 2; (b) a polynucleotide which codes for a polypeptide which comprises an amino acid sequence which is identical to the extent of at least 70% to the amino acid sequence of SEQ ID NO: 2; (c) a polynucleotide which is complementary to the polynucleotides of (a) or (b); and (d) a polynucleotide comprising at least 15 successive nucleotides of the polynucleotide sequence of (a), (b), or (c), and a process for the fermentative preparation of L-amino acids using coryneform bacteria in which at least the sigE gene is present in enhanced form, and the use of polynucleotides which comprise the sequence according to the invention as hybridization probes.

1 - 20 . (canceled)  
     
     
         21 . An isolated nucleic acid comprising a nucleotide sequence selected from the group consisting of: 
 (a) a nucleotide sequence as set forth in SEQ ID NO: 1;    (b) a nucleotide sequence encoding the polypeptide as set forth in SEQ ID NO: 2; and    (c) a nucleotide sequence fully complementary to (a) or (b).    
     
     
         22 . A vector comprising the nucleic acid of  claim 21 .  
     
     
         23 . A bacterium comprising the vector of  claim 22 , wherein said bacterium is an  E. coli  or coryneform bacterium.  
     
     
         24 . A recombinant  Corynebacterium glutamicum  comprising an overexpressed polynucleotide encoding a polypeptide having the amino acid sequence of SEQ ID NO: 2, wherein overexpression is achieved by increasing the copy number of said polynucleotide or by operably linking promoter to said polynucleotide.  
     
     
         25 . An isolated nucleic acid that encodes a polypeptide comprising an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO: 2, wherein said polypeptide has a biological activity of a sigma factor E protein.  
     
     
         26 . A vector comprising the nucleic acid of  claim 25 .  
     
     
         27 . A host cell comprising the vector of  claim 26 .  
     
     
         28 . An isolated polynucleotide primer or probe of a nucleic acid fragment, wherein said fragment consists of at least 30 consecutive nucleotides from SEQ ID NO: 1 or the full complement of said fragment.  
     
     
         29 . An isolated primer or probe consisting of a nucleic acid fragment, wherein said fragment consists of at least 40 consecutive nucleotides from SEQ ID NO: 1 or the full complement of said fragment.  
     
     
         30 . The shuttle vector pEC-T 18mob2sigEexp having 
 (a) a 1930 bp nucleic acid fragment of SEQ ID NO: 4 which harbors the  C. glutamicum  gene; and    (b) a restriction map as set forth in  FIG. 2 .    
     
     
         31 . The shuttle vector of  claim 30 , wherein the vector has been deposited in  Corynebacterium glutamicum  strain DSM5715/pEC-T18mob2sigEexp under accession no. DSM 14229.  
     
     
         32 . An isolated polynucleotide comprising nucleotides 236 to 907 of SEQ ID NO: 1.