Bacillus licheniformis chromosome

Abstract

The present invention relates to an isolated polynucleotide of the complete chromosome of Bacillus licheniformis . The present invention also relates to isolated genes of the chromosome of Bacillus licheniformis which encode biologically active substances and to nucleic acid constructs, vectors, and host cells comprising the genes as well as methods for producing biologically active substances encoded by the genes and to methods of using the isolated genes of the complete chromosome of Bacillus licheniformis.

1 . An isolated gene which encodes a polypeptide having aspartate 1-decarboxylase activity, selected from the group consisting of:
 (a) a gene comprising a nucleotide sequence having at least 90% sequence identity with the nucleotide sequence of SEQ ID NO: 2275; and   (b) a gene comprising a nucleotide sequence which hybridizes under high stringency conditions with the nucleotide sequence of SEQ ID NO: 2275, or the complementary strand thereof, wherein high stringency conditions are defined as prehybridization and hybridization at 42° C. in 5×SSPE, 0.3% SDS, 200 μg/ml sheared and denatured salmon sperm DNA, and 50% formamide, and washing three times each for 15 minutes using 2×SSC, 0.2% SDS at 65° C.   
     
     
         2 . The isolated gene of  claim 1 , which comprises a nucleotide sequence having at least 90% sequence identity with SEQ ID NO: 2275. 
     
     
         3 . The isolated gene of  claim 1 , which comprises a nucleotide sequence having at least 95% sequence identity with SEQ ID NO: 2275. 
     
     
         4 . The isolated gene of  claim 1 , which comprises a nucleotide sequence having at least 97% sequence identity with SEQ ID NO: 2275. 
     
     
         5 . The isolated gene of  claim 1 , which comprises the nucleotide sequence of SEQ ID NO: 2275. 
     
     
         6 . The isolated gene of  claim 1 , which hybridizes under high stringency conditions with the nucleotide sequence of SEQ ID NO: 2275, or the complementary strand thereof, wherein high stringency conditions are defined as prehybridization and hybridization at 42° C. in 5×SSPE, 0.3% SDS, 200 μg/ml sheared and denatured salmon sperm DNA, and 50% formamide, and washing three times each for 15 minutes using 2×SSC, 0.2% SDS at 65° C. 
     
     
         7 . The isolated gene of  claim 1 , which hybridizes under very high stringency conditions with the nucleotide sequence of SEQ ID NO: 2275, or the complementary strand thereof, wherein very high stringency conditions are defined as prehybridization and hybridization at 42° C. in 5×SSPE, 0.3% SDS, 200 μg/ml sheared and denatured salmon sperm DNA, and 50% formamide, and washing three times each for 15 minutes using 2×SSC, 0.2% SDS at 70° C. 
     
     
         8 . The isolated gene of  claim 1 , which is contained in  Bacillus licheniformis  ATCC 14580. 
     
     
         9 . A nucleic acid construct comprising the isolated gene of  claim 1  operably linked to one or more control sequences that direct the production of the polypeptide in a host cell. 
     
     
         10 . A recombinant expression vector comprising the nucleic acid construct of  claim 9 . 
     
     
         11 . A recombinant host cell comprising the nucleic acid construct of  claim 9 . 
     
     
         12 . An isolated gene which encodes a polypeptide having aspartate 1-decarboxylase activity, wherein the polypeptide has at least 90% sequence identity with the amino acid sequence of SEQ ID NO: 6472. 
     
     
         13 . The isolated gene of  claim 12 , wherein the polypeptide has at least 95% sequence identity with the amino acid sequence of SEQ ID NO: 6472. 
     
     
         14 . The isolated gene of  claim 12 , wherein the polypeptide has at least 97% sequence identity with the amino acid sequence of SEQ ID NO: 6472. 
     
     
         15 . The isolated gene of  claim 12 , wherein the polypeptide comprises the amino acid sequence of SEQ ID NO: 6472. 
     
     
         16 . A nucleic acid construct comprising the isolated gene of  claim 12  operably linked to one or more control sequences that direct the production of the polypeptide in an expression host. 
     
     
         17 . A recombinant expression vector comprising the nucleic acid construct of  claim 16 . 
     
     
         18 . A recombinant host cell comprising the nucleic acid construct of  claim 16 . 
     
     
         19 . A recombinant host cell comprising a polynucleotide which encodes a polypeptide having aspartate 1-decarboxylase activity and at least 90% sequence identity with the amino acid sequence of SEQ ID NO: 6472, wherein the polynucleotide is operably linked to a promoter heterologous to the host cell. 
     
     
         20 . The recombinant host cell  claim 19 , wherein the polypeptide has at least 95% sequence identity with the amino acid sequence of SEQ ID NO: 6472.