Nucleotide sequences encoding the glk-gene

Abstract

The invention relates to an isolated polynucleotide containing a polynucleotide sequence selected from the group comprising: a) a polynucleotide that is at least 70% identical to a polynucleotide encoding a polypeptide that contains the amino acid sequence of SEQ ID NO:2, b) a polynucleotide encoding a polypeptide that contains an amino acid sequence that is at least 70% identical to the amino acid sequence of SEQ ID NO:2, c) a polynucleotide that is complementary to the polynucleotides of a) or b), and d) a polynucleotide containing at least 15 successive bases of the polynucleotide sequence of a), b) or c), and processes for the fermentative production of L-amino acids by enhancement of the glk-gene coding for the enzyme glucokinase.

What is claimed is:  
     
         1 . An isolated polynucleotide obtained from coryneform bacteria, containing a polynucleotide sequence selected from the group consisting of 
 a) a polynucleotide that is at least 70% identical to a polynucleotide coding for a polypeptide that contains the amino acid sequence of SEQ ID NO:2,    b) a polynucleotide encoding a polypeptide, that contains an amino acid sequence that is at least 70% identical to the amino acid sequence of SEQ ID NO:2,    c) a polynucleotide that is complementary to the polynucleotides of a) or b), and    d) a polynucleotide containing at least 15 successive nucleotides of the polynucleotide sequence of a), b) or c).    
     
     
         2 . The polynucleotide according to  claim 1 , wherein the polynucleotide is a DNA replicable in coryneform bacteria.  
     
     
         3 . The polynucleotide according to  claim 2  that is a recombinant DNA.  
     
     
         4 . The polynucleotide according to  claim 1 , wherein the polynucleotide is an RNA.  
     
     
         5 . The replicable DNA according to  claim 2 , containing 
 (i) the nucleotide sequence shown in SEQ ID NO:1, or    (ii) at least one sequence that corresponds to the sequence (i) within the region of degeneracy of the genetic code, or    (iii) at least one sequence that hybridises with the sequence that is complementary to the sequence (i) or (ii), and optionally    (iv) functionally neutral sense mutations in (i).    
     
     
         6 . The polynucleotide sequence according to  claim 2 , encoding a polypeptide that contains the amino acid sequence shown in SEQ ID NO:2.  
     
     
         7 . A vector containing a polynucleotide sequence according to  claim 1 .  
     
     
         8 . Coryneform bacteria containing a vector according to  claim 7 .  
     
     
         9 . A process for the fermentative production of L-amino acids, comprising the following steps: 
 a) fermentation of the coryneform bacteria producing the L-amino acid, in which at least the gene coding for the enzyme glucokinase is enhanced,    b) enrichment of the L-amino acid in the medium or in the cells of the bacteria, and    c) isolation of the L-amino acid.    
     
     
         10 . The process according to  claim 9  in which the gene is enhanced by overexpression.  
     
     
         11 . The process according to  claim 9 , wherein bacteria are used in which, additionally, further genes of the biosynthesis pathway of the desired L-amino acid are enhanced.  
     
     
         12 . The process according to  claim 9 , wherein bacteria are used in which the metabolic pathways that reduce the formation of the L-amino acid are at least partially switched off.  
     
     
         13 . The process according to  claim 9 , wherein a strain transformed with a plasmid vector is used, and said plasmid vector carries the nucleotide sequence of the gene encoding the enzyme glucokinase.  
     
     
         14 . The process according to one of  claims 9  to  13 , wherein coryneform bacteria are used that produce L-lysine.  
     
     
         15 . The process according to  claim 11 , wherein the dapA-gene encoding dihydrodipicolinate synthase is simultaneously overexpressed.  
     
     
         16 . The process according to  claim 11 , wherein a lysC-gene encoding a feedback-resistant aspartate kinase is simultaneously overexpressed.  
     
     
         17 . The process according to  claim 11 , wherein the gap-gene encoding gyceraldehyde-3-phosphate dehydrogenase is simultaneously overexpressed.  
     
     
         18 . The process according to  claim 11 , wherein the tpi-gene encoding triosephosphate isomerase is simultaneously overexpressed.  
     
     
         19 . The process according to  claim 11 , wherein the pgk-gene encoding 3-phosphate glycerate kinase is simultaneously overexpressed.  
     
     
         20 . The process according to  claim 11 , wherein the pyc-gene coding for pyruvate carboxylase is simultaneously overexpressed.  
     
     
         21 . The process according to  claim 11 , wherein the mqo-gene encoding malate-quinone oxidoreductase is simultaneously overexpressed.  
     
     
         22 . The process according to  claim 11 , wherein the lysE-gene encoding lysine export is simultaneously overexpressed.  
     
     
         23 . The process according to  claim 12 , wherein the pgi-gene encoding glucose-6-phosphate isomerase is simultaneously attenuated.  
     
     
         24 . The process according to  claim 12 , wherein the pck-gene encoding phosphoenolpyruvate carboxykinase is simultaneously attenuated.