Summary: Degenerate PCR primers were designed from the N-terminal amino acid sequence of a glutamyl aminopeptidase (PepA) from . These primers were used to screen a lambda library for clones containing the gene () encoding PepA. The DNA sequence of a 2-1 kb fragment containing was determined. The sequence revealed the presence of one complete and two incomplete open reading frames (ORFs). The complete ORF encodes a putative protein of 353 amino acids with a predicted N-terminal sequence identical to that determined for purified PepA. The gene was subcloned on an plasmid vector and production of active PepA was confirmed by means of a zymogram. Mutants of in which the gene was inactivated grew to normal cell densities in milk but exhibited a reduced growth rate during the exponential phase. Thus whilst PepA is required for optimal growth it is not essential.


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