Novel nucleotide sequences coding for the glbO gene

Abstract

Isolated polynucleotides containing at least one nucleic acid sequence selected from the group a) polynucleotides which is at least 70% identical to a polynucleotide, b) b) polynucleotide which codes for a polypeptide which contains an amino acid sequence which is at least 70% identical to an amino acid sequence o, c) polynucleotide which is complementary to the polynucleotides of a) or b), and d) polynucleotide containing at least 15 successive nucleotides of the polynucleotide sequence of a), b) or c), and a method for the fermentative production of L-amino acids with amplification of the glb0 gene which codes for the hemoglobin-like protein and the use of the above polynucleotides as a primer or hybridization probe.

What is claimed is:  
     
         1 . An isolated polynucleotide from coryneform bacteria containing one of the following polynucleotide sequences: 
 a) a polynucleotide which is at least 70% identical to a polynucleotide which codes for a polypeptide containing the amino acid sequence of SEQ ID NO: 2,    b) a polynucleotide which codes for a polypeptide which contains an amino acid sequence which is at least 70% identical 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 containing at least 15 consecutive nucleotides of the polynucleotide sequence of a), b) or c).    
     
     
         2 . The polynucleotide of  claim 1 , wherein the polynucleotide is a recombinant DNA replicable in coryneform bacteria.  
     
     
         3 . The polynucleotide of  claim 1 , wherein the polynucleotide is an RNA.  
     
     
         4 . The polynucleotide of  claim 2 , wherein the replicable DNA comprises: 
 (i) the nucleotide sequence shown in SEQ ID NO: 1, or    (ii) at least one sequence which matches the sequence of (i) within the degeneration range of the genetic code, or    (iii) at least one sequence which hybridizes with the complementary sequence to sequence (i) or (ii), or    (iv) functionally neutral sense mutations in (i).    
     
     
         5 . The polynucleotide of  claim 2  and having a sequence which codes for a polypeptide which comprises the amino acid sequence as shown in SEQ ID NO: 2.  
     
     
         6 . A vector containing the polynucleotide sequence of  claim 1 .  
     
     
         7 . A coryneform bacterium containing the vector as claimed in  claim 6 .  
     
     
         8 . A method for the fermentative production of L-amino acids, comprising: 
 a) fermenting L-amino acid producing coryneform bacteria in which at least the gene which codes for the hemoglobin-like protein is amplified.    
     
     
         9 . The method of  claim 8 , wherein bacteria are used in which further genes of the biosynthetic pathway of the desired L-amino acid are additionally amplified.  
     
     
         10 . The method of  claim 8 , wherein bacteria are used in which the metabolic pathways which reduce the formation of the L-amino acid are at least partially suppressed.  
     
     
         11 . The method of  claim 8 , wherein bacteria transformed with a plasmid vector are used and the plasmid vector bears the nucleotide sequence of the gene which codes for the hemoglobin-like protein.  
     
     
         12 . The method of  claim 8 , wherein coryneform bacteria are used which produce L-lysine.  
     
     
         13 . The method  claim 9 , wherein one or more genes selected from the group consisting of the dapA gene which codes for dihydropicolinate synthase,the pyc gene which codes for pyruvate carboxylase,the tpi gene which codes for triosephosphate isomerase, the gap gene which codes for glyceraldehyde-3-phosphate dehydrogenase,the pgk gene which codes for 3-phosphoglycerate kinase, and the lysE gene which codes for lysine export is/are simultaneously amplified.  
     
     
         14 . The method of  claim 10 , wherein bacteria are fermented for the production of L-lysine in which one or more of the genes selected from the group consisting of the pck gene which codes for phosphoenolpyruvate carboxykinase and the pgi gene which codes for glucose-6-phosphate isomerase is/are simultaneously attenuated.  
     
     
         15 . The method of  claim 8 , wherein microorganisms of the genus  Corynebacterium glutamicum  are used.  
     
     
         16 . The method of  claim 8  further comprising: 
 b. accumulating L-amino acid in the medium or in the cells of the bacteria.  
 
     
     
         17 . The method of  claim 16  further comprising: 
 c. isolating the L-amino acid.  
 
     
     
         18 . A primer which comprises a polynucleotide sequence of  claim 1 , wherein the primer is used for the production of DNA of genes which code for the gene product of Glb0 by the polymerase chain reaction.  
     
     
         19 . A hybridization probe comprising a polynucleotide sequence of  claim 1.