F0F1-ATPase and DNA encoding the same

Abstract

Provided are a protein complex having the F 0 F 1 -ATPase activity; a DNA encoding the protein complex; a method for producing the protein complex, using the DNA; and a method for producing nucleoside 5′-triphosphate, using the protein. The present invention further provides a recombinant DNA with the DNA inserted therein; a transformant carrying the recombinant DNA; and a method for producing a protein complex, using the transformant.

What is claimed is:  
     
         1 . A protein selected from the group consisting of the following proteins (a) to (c): 
 (a) a protein having the amino acid sequence represented by SEQ ID NO: 1;    (b) a protein having a modified one of the amino acid sequence represented by SEQ ID NO: 1, where one or more amino acids are deleted, substituted or added, and exerting the F 0 F 1 -ATPase activity when the protein forms a complex with all the individual proteins having the amino acid sequences represented by each of SEQ ID NOS: 2 to 8; and    (c) a protein having an amino acid sequence having 70% or more identical to the amino acid sequence represented by SEQ ID NO: 1 and exerting the F 0 F 1 -ATPase activity when the protein forms a complex with all the individual proteins having the amino acid sequences represented by each of SEQ ID NOS: 2 to 8.    
     
     
         2 . A protein selected from the group consisting of the following proteins (a) to (c): 
 (a) a protein having the amino acid sequence represented by SEQ ID NO: 2;    (b) a protein having a modified one of the amino acid sequence represented by SEQ ID NO: 2, where one or more amino acids are deleted, substituted or added, and exreting the F 0 F 1 -ATPase activity when the protein forms a complex with all the individual proteins having the amino acid sequences represented by each of SEQ ID NO: 1 and SEQ ID NO: 3 to 8; and    (c) a protein having an amino acid sequence having 70% or more identical to the amino acid sequence represented by SEQ ID NO: 2 and exerting the F 0 F 1 -ATPase activity when the protein forms a complex with all the individual proteins having the amino acid sequences represented by each of SEQ ID NO: 1 and SEQ ID NOS: 3 to 8.    
     
     
         3 . A protein selected from the group consisting of the following proteins (a) to (c): 
 (a) a protein having the amino acid sequence represented by SEQ ID NO: 3;    (b) a protein having a modified one of the amino acid sequence represented by SEQ ID NO: 3, where one or more amino acids are deleted, substituted or added, and exerting the F 0 F 1 -ATPase activity when the protein forms a complex with all the proteins having the individual amino acid sequences represented by each of SEQ ID NOS: 1 and 2 and SEQ ID NOS: 4 to 8; and    (c) a protein having an amino acid sequence having 70% or more identical to the amino acid sequence represented by SEQ ID NO: 3 and exerting the F 0 F 1 -ATPase activity when the protein forms a complex with all the individual proteins having the amino acid sequences represented by each of SEQ ID NOS: 1 and 2 and SEQ ID NOS: 4 to 8.    
     
     
         4 . A protein selected from the group consisting of the following proteins (a) to (c): 
 (a) a protein having the amino acid sequence represented by SEQ ID NO: 4;    (b) a protein having a modified one of the amino acid sequence represented by SEQ ID NO: 4, where one or more amino acids are deleted, substituted or added, and exerting the F 0 F 1 -ATPase activity when the protein forms a complex with all the individual proteins having the amino acid sequences represented by each of SEQ ID NOS: 1 to 3 and SEQ ID NOS: 5 to 8; and    (c) a protein having an amino acid sequence having 70% or more identical to the amino acid sequence represented by SEQ ID NO: 4 and exerting the F 0 F 1 -ATPase activity when the protein forms a complex with all the individual proteins having the amino acid sequences represented by each of SEQ ID NO: 1 to 3 and SEQ ID NOS: 5 to 8.    
     
     
         5 . A protein selected from the group consisting of the following proteins (a) to (c): 
 (a) a protein having the amino acid sequence represented by SEQ ID NO: 5;    (b) a protein comprising a modified one of the amino acid sequence represented by SEQ ID NO: 5, where one or more amino acids are deleted, substituted or added, and exerting the F 0 F 1 -ATPase activity when the protein forms a complex with all the individual proteins having the amino acid sequences represented by each of SEQ ID NOS: 1 to 4 and SEQ ID NOS: 6 to 8; and    (c) a protein having an amino acid sequence having 70% or more identical to the amino acid sequence represented by SEQ ID NO: 5 and exerting the F 0 F 1 -ATPase activity when the protein forms a complex with all the individual proteins having the amino acid sequences represented by each of SEQ ID NOS: 1 to 4 and SEQ ID NOS: 6 to 8.    
     
     
         6 . A protein selected from the group consisting of the following proteins (a) to (c): 
 (a) a protein having the amino acid sequence represented by SEQ ID NO: 6;    (b) a protein having a modified one of the amino acid sequence represented by SEQ ID NO: 6,wherein one or more amino acids are deleted, substituted or added, and which can exert the F 0 F 1 -ATPase activity when the protein forms a complex with all the individual proteins having the amino acid sequences represented by each of SEQ ID NOS: 1 to 5 and SEQ ID NOS: 7 and 8; and    (c) a protein having an amino acid sequence having 70% or more identical to the amino acid sequence represented by SEQ ID NO: 6 and exerting the F 0 F 1 -ATPase activity when the protein forms a complex with all the individual proteins having the amino acid sequences represented by each of SEQ ID NOS: 1 to 5 and SEQ ID NOS: 7 and 8.    
     
     
         7 . A protein selected from the group consisting of the following proteins (a) to (c): 
 (a) a protein having the amino acid sequence represented by SEQ ID NO: 7;    (b) a protein having a modified one of the amino acid sequence represented by SEQ ID NO: 7, where one or more amino acids are deleted, substituted or added, and exerting the F 0 F 1 -ATPase activity when the protein forms a complex with all the individual proteins having the amino acid sequences represented by each of SEQ ID NOS: 1 to 6 and SEQ ID NO: 8; and    (c) a protein having an amino acid sequence having 70% or more identity to the amino acid sequence represented by SEQ ID NO: 7 and exerting the F 0 F 1 -ATPase activity when the protein forms a complex with all the individual proteins having the amino acid sequences represented by each of SEQ ID NOS: 1 to 6 and SEQ ID NO: 8.    
     
     
         8 . A protein selected from the group consisting of the following proteins (a) to (c): 
 (a) a protein having the amino acid sequence represented by SEQ ID NO: 8;    (b) a protein having a modified one of the amino acid sequence represented by SEQ ID NO: 8, where one or more amino acids are deleted, substituted or added, and exerting the F 0 F 1 -ATPase activity when the protein forms a complex with all the individual proteins having the amino acid sequences represented by each of SEQ ID NOS: 1 to 7; and    (c) a protein having an amino acid sequence having 70% or more identity to the amino acid sequence represented by SEQ ID NO: 8 and exerting the F 0 F 1 -ATPase activity when the protein forms a complex with all the individual proteins comprising the amino acid sequences represented by each of SEQ ID NOS: 1 to 7.    
     
     
         9 . A protein complex comprising eight proteins respectively selected from the eight groups as defined by each of  claims 1  to  8 .  
     
     
         10 . A DNA encoding any one of the proteins according to  claims 1  to  8 .  
     
     
         11 . A DNA selected from the group consisting of the following DNAs (a) and (b): 
 (a) a DNA having the nucleotide sequence represented by SEQ ID NO:9; and    (b) a DNA hybridizing with the DNA under stringent conditions and encoding a protein exerting the F 0 F 1 -ATPase activity when the protein forms a complex with all the individual proteins having the amino acid sequences represented by each of SEQ ID NOS: 2 to 8.    
     
     
         12 . A DNA selected from the group consisting of the following DNAs (a) and (b): 
 (a) a DNA having the nucleotide sequence represented by SEQ ID NO: 10; and    (b) a DNA hybridizing with the DNA under stringent conditions and encoding a protein exerting the F 0 F 1 -ATPase activity when the protein forms a complex with all the individual proteins having the amino acid sequences represented by each of SEQ ID NO: 1 and SEQ ID NOS: 3 to 8.    
     
     
         13 . A DNA selected from the group consisting of the following DNAs (a) and (b): 
 (a) a DNA having the nucleotide sequence represented by SEQ ID NO: 11; and    (b) a DNA hybridizing with the DNA under stringent conditions and encoding a protein exerting the F 0 F 1 -ATPase activity when the protein forms a complex with all the individual proteins having the amino acid sequences represented by each of SEQ ID NOS: 1 and 2 and SEQ ID NOS: 4 to 8.    
     
     
         14 . A DNA selected from the group consisting of the following DNAs (a) and (b): 
 (a) a DNA having the nucleotide sequence represented by SEQ ID NO:12; and    (b) a DNA hybridizing with the DNA under stringent conditions and encoding a protein exerting the F 0 F 1 -ATPase activity when the protein forms a complex with all the individual proteins having the individual amino acid sequences represented by each of SEQ ID NOS: 1 to 3 and SEQ ID NOS: 5 to 8.    
     
     
         15 . A DNA selected from the group consisting of the following DNAs (a) and (b): 
 (a) a DNA having the nucleotide sequence represented by SEQ ID NO:13; and    (b) a DNA hybridizing with the DNA under stringent conditions and encoding a protein exerting the F 0 F 1 -ATPase activity when the protein forms a complex with all the individual proteins having the amino acid sequences represented by each of SEQ ID NOS: 1 to 4 and SEQ ID NOS: 6 to 8.    
     
     
         16 . A DNA selected from the group consisting of the following DNAs (a) and (b): 
 (a) a DNA having the nucleotide sequence represented by SEQ ID NO: 14; and    (b) a DNA hybridizing with the DNA under stringent conditions and encoding a protein exerting the F 0 F 1 -ATPase activity when the protein forms a complex with all the individual proteins having the amino acid sequences represented by each of SEQ ID NOS: 1 to 5 and SEQ ID NOS: 7 and 8.    
     
     
         17 . A DNA selected from the group consisting of the following DNAs (a) and (b): 
 (a) a DNA having the nucleotide sequence represented by SEQ ID NO: 15; and    (b) a DNA hybridizing with the DNA under stringent conditions and encoding a protein exerting the F 0 F 1 -ATPase activity when the protein forms a complex with all the individual proteins having the amino acid sequences represented by each of SEQ ID NOS: 1 to 6 and SEQ ID NO: 8.    
     
     
         18 . A DNA selected from the group consisting of the following DNAs (a) and (b): 
 (a) a DNA having the nucleotide sequence represented by SEQ ID NO: 16; and    (b) a DNA hybridizing with the DNA under stringent conditions and encoding a protein exerting the F 0 F 1 -ATPase activity when the protein forms a complex with all the individual proteins having the amino acid sequences represented by each of SEQ ID NOS: 1 to 7.    
     
     
         19 . A DNA comprising the eight DNAs respectively selected from the eight groups as defined by each of  claims 11  to  18 .  
     
     
         20 . A DNA having the nucleotide sequences represented by SEQ ID NOS: 9 to 16.  
     
     
         21 . A DNA having the nucleotide sequence represented by SEQ ID NO: 21.  
     
     
         22 . The DNA according to any one of  claims 10  to  21 , where the DNA is derived from a microorganism belonging to the genus Corynebacterium.  
     
     
         23 . The DNA according to any one of  claims 10  to  21 , where the DNA is derived from a microorganism of the species  Corynebacterium ammoniagenes.    
     
     
         24 . A recombinant DNA constructed by inserting the DNA according to any one of  claims 10  to  18  into a vector.  
     
     
         25 . A recombinant DNA constructed by inserting the DNA according to any one of  claims 19  to  21  into a vector.  
     
     
         26 . A transformant obtained by transformation of a host cell with the recombinant DNA according to  claim 24  or  25 .  
     
     
         27 . A transformant according to  claim 26 , where the host cell is a microorganism of the species  Escherichia coli, Corynebacterium glutamicum  or  Corynebacterium ammoniagenes.    
     
     
         28 . A method for producing a protein according to any one of  claims 1  to  8 , which comprises culturing a transformant obtained by transformation of a host cell with the recombinant DNA according to  claim 24  in a culture medium, so as to allow the protein according to any one of  claims 1  to  8  to be expressed and accumulated in the culture and harvesting the protein from the culture.  
     
     
         29 . A method for producing a protein complex having the F 0 F 1 -ATPase activity, which comprises culturing a transformant obtained by transformation of a host cell with the recombinant DNA according to  claim 25  in a culture medium, so as to allow a protein complex having the F 0 F 1 -ATPase activity to be expressed and accumulated in the culture and recovering the protein complex from the culture.  
     
     
         30 . A method for producing nucleoside 5′-triphosphate, which comprises by use of a culture of a transformant obtained by transformation of a host cell with the recombinant DNA according to  claim 25  or a treated product of the culture as an enzyme source, allowing the enzyme source and a precursor of nucleoside 5′-triphosphate to co-exist with each other in an aqueous medium to generate and accumulate the nucleoside 5′-triphosphate and recovering the nucleoside 5′-triphosphate from the aqueous medium.  
     
     
         31 . The method according to claim  30 , where the precursor of nucleoside 5′-triphosphate is adenine, guanine, uracil, cytosine, hypoxanthine, adenosine, guanosine, uridine, cytidine, inosine, adenosine 5′-monophosphate, guanosine 5′-monophosphate, uridine 5′-monophosphate, cytidine 5′-monophosphate or inosine 5′-monophosphate.  
     
     
         32 . The method according to claim  30 , where the nucleoside 5′-triphosphate is adenosine 5′-triphosphate, guanosine 5′-triphosphate, uridine 5′-triphosphate or cytidine 5′-triphosphate.