A NOVEL AMIDOHYDROLASE GENE FROM BACILLUS-SUBTILIS CLONING - DNA-SEQUENCE ANALYSIS AND MAP POSITION OF AMHX

The nucleotide sequence of a new Bacillus subtilis gene (amhX) was det ermined that encodes a protein (AmhX) with strong sequence identity to amidohydrolases from both plant and bacterial species and a carboxype ptidase from the archaeon Sulfolobus sulfataricus. The amhX gene encod es a hydrophilic polypeptide of 383 amino acids with a molecular mass of 41.5 kDa. The amhX gene was overexpressed in E. coli by using the T 7 RNA polymerase/promoter system and the transcription initiation site s for the amhX mRNAs in B. subtilis were determined by primer extensio n analysis. Chromosomal amhX mutations were constructed by marker repl acement and the amhX gene was positioned at 25 degrees on the genetic and physical map of the B. subtilis chromosome.