Carbamoyl-phosphate synthetase gene of coryneform bacteria and method for producing l-arginine

Abstract

A DNA fragment which encodes a polypeptide defined in the following (a) or (b), and a polypeptide defined in the following (c) or (d): (a) a polypeptide which has at least the amino acid sequence of the amino acid numbers 50 to 393 in SEQ ID NO: 2 shown in Sequence Listing, (b) a polypeptide which has at least the amino acid sequence of the amino acid numbers 50 to 393 in SEQ ID NO: 2 shown in Sequence Listing including substitution, deletion, insertion, addition, or inversion of one or several amino acids, and can constitute a protein having a carbamoyl-phosphate synthetase activity with a large subunit of carbamoyl-phosphate synthetase having the amino acid sequence of SEQ ID NO: 3, (c) a polypeptide which has the amino acid sequence of SEQ ID NO: 3 shown in Sequence Listing, (d) a polypeptide which has the amino acid sequence of SEQ ID NO: 3 shown in Sequence Listing including substitution, deletion, insertion, addition, or inversion of one or several amino acids, and can constitute a protein having a carbamoyl-phosphate synthetase activity with a small subunit of carbamoyl-phosphate synthetase having the amino acid sequence of the amino acid numbers 50 to 393 in SEQ ID NO: 2.

What is claimed is:  
     
         1 . A DNA fragment which encodes a polypeptide defined in the following (A) or (B): 
 (A) a polypeptide which has an amino acid sequence comprises at least the amino acid numbers 50 to 393 of the amino acid sequence of SEQ ID NO: 2,    (B) a polypeptide which has an amino acid sequence comprises at least the amino acid numbers 50 to 393 of the amino acid sequence of SEQ ID NO: 2 including substitution, deletion, insertion, addition, or inversion of one or several amino acids, and can constitute a protein having a carbamoyl-phosphate synthetase activity with a large subunit of carbamoyl-phosphate synthetase comprising the amino acid sequence of SEQ ID NO: 3.    
     
     
         2 . A DNA fragment which encodes a polypeptide defined in the following (C) or (D): 
 (C) a polypeptide which comprises the amino acid sequence of SEQ ID NO: 3,    (D) a polypeptide which comprises the amino acid sequence of SEQ ID NO: 3 including substitution, deletion, insertion, addition, or inversion of one or several amino acids, and can constitute a protein having a carbamoyl-phosphate synthetase activity with a small subunit of carbamoyl-phosphate synthetase having an amino acid sequence comprises at least the amino acid numbers 50 to 393 of the amino acid sequence of SEQ ID NO: 2.    
     
     
         3 . A DNA fragment encoding a polypeptide which comprises the amino acid sequence of SEQ ID NO: 3 including substitution, deletion, insertion, addition, or inversion of one or several amino acids, and can constitute a protein having a carbamoyl-phosphate synthetase activity.  
     
     
         4 . A DNA fragment which encodes a polypeptide defined in the following (a) or (b), and a polypeptide defined in the following (c) or (d): 
 (a) a polypeptide which has an amino acid sequence comprising at least the amino acid numbers 50 to 393 in SEQ ID NO: 2,    (b) a polypeptide which has an amino acid sequence comprising at least the amino acid numbers 50 to 393 in SEQ ID NO: 2 including substitution, deletion, insertion, addition, or inversion of one or several amino acids, and can constitute a protein having a carbamoyl-phosphate synthetase activity with a large subunit of carbamoyl-phosphate synthetase comprising the amino acid sequence of SEQ ID NO: 3,    (c) a polypeptide which comprises the amino acid sequence of SEQ ID NO: 3,    (d) a polypeptide which comprises the amino acid sequence of SEQ ID NO: 3 including substitution, deletion, insertion, addition, or inversion of one or several amino acids, and can constitute a protein having a carbamoyl-phosphate synthetase activity with a small subunit of carbamoyl-phosphate synthetase having an amino acid sequence comprising the amino acid numbers 50 to 393 in SEQ ID NO: 2.    
     
     
         5 . The DNA fragment according to  claim 1 , which has a nucleotide sequence comprising at least the nucleotide numbers 430 to 1461 in the nucleotide sequence of SEQ ID NO: 1.  
     
     
         6 . The DNA fragment according to  claim 2 , which has a nucleotide sequence comprising at least the nucleotide numbers 1756 to 4809 in the nucleotide sequence of SEQ ID NO: 1.  
     
     
         7 . The DNA fragment according to  claim 3 , which has a nucleotide sequence comprising at least the nucleotide numbers 430 to 4809 in the nucleotide sequence of SEQ ID NO: 1.  
     
     
         8 . A protein which comprises a polypeptide defined in the following (a) or (b), and a polypeptide defined in the following (c) or (d): 
 (a) a polypeptide which has an amino acid sequence comprising at least the amino acid numbers 50 to 393 in SEQ ID NO: 2,    (b) a polypeptide which has an amino acid sequence comprising at least the amino acid numbers 50 to 393 in SEQ ID NO: 2 including substitution, deletion, insertion, addition, or inversion of one or several amino acids, and can constitute a protein having a carbamoyl-phosphate synthetase activity with a large subunit of carbamoyl-phosphate synthetase comprising the amino acid sequence of SEQ ID NO: 3,    (c) a polypeptide which comprises the amino acid sequence of SEQ ID NO: 3,    (d) a polypeptide which comprises the amino acid sequence of SEQ ID NO: 3 including substitution, deletion, insertion, addition, or inversion of one or several amino acids, and can constitute a protein having a carbamoyl-phosphate synthetase activity with a small subunit of carbamoyl-phosphate synthetase having an amino acid sequence comprising at least the amino acid numbers 50 to 393 in SEQ ID NO: 2.    
     
     
         9 . A coryneform bacterium which is transformed with a DNA fragment according to any one of  claims 1  to  7 .  
     
     
         10 . A microorganism which has enhanced intracellular carbamoyl-phosphate synthetase activity, and has L-arginine productivity.  
     
     
         11 . The microorganism according to  claim 10 , wherein the enhanced intracellular carbamoyl-phosphate synthetase activity is obtained by increasing copy number of DNA encoding carbamoyl-phosphate synthetase of the microorganism, or by modifying an expression regulation sequence so that expression of the gene encoding carbamoyl-phosphate synthetase in the cell should be enhanced.  
     
     
         12 . The microorganism according to  claim 11 , wherein the DNA is a DNA fragment according to any one of  claims 1  to  7 .  
     
     
         13 . The microorganism according to  claim 12 , which is a coryneform bacterium.  
     
     
         14 . A method for producing of L-arginine, comprising the steps of culturing a coryneform bacterium according to any one of  claims 10  to  13  in a medium to produce and accumulate L-arginine in the medium, and collecting the L-arginine from the medium.