CLONING AND NUCLEOTIDE-SEQUENCE OF GAMMA-POLYGLUTAMATE PRODUCTION STIMULATING FACTOR ON BACILLUS-SUBTILIS (NATTO) PLASMID, PUH1

The gene coding for gamma-polyglutamate production stimulating factor (psf) from Bacillus subtilis (natto) plasmid pUHl was cloned and seque nced in Bacillus subtilis host. The activity of gamma-glutamyltranspep tidase (gamma-GTP) in B. subtilis host, which was introduced the psf g ene was relatively high level, while its activity in E. coli host coul d not be detected. The nucleotide sequence of psf gene was determined and an open reading frame encoding a polypeptide composed of 420 amino acid residues (Mr, 49, 356) was identified. The putative -35 and -10 sequences, TTCAAA and TATTAT, were observed as the consensus sequence for the promoter recognized by the sigma(43) RNA polymerase of B. subt ilis, and the ribosome binding site, whose sequence was AACGAG, was co mplementary to the binding sequence of B. subtilis 16S rRNA except for one base. The amino acid sequence of psf with the segment of putative protein C403 of staphylococcal plasmid pE194 indicates homology, wher eas that with E. coli and mammalian gamma-GTPs does not show any simil arity at all.