The upstream region of the -acetylmuramoyl-L-alanine amidase gene (; a major autolysin) was cloned into by chromosome walking. Sequencing of the region showed the presence of two open reading frames, one (designated as ) which starts at a UUG codon and encodes a polypeptide of 705 amino acids with an of 76725, and the other (designated as ), upstream of , comprising 102 amino acids and having a signal sequence characteristic of a lipoprotein. Purification of the CwbA protein and determination of its N-terminal amino acid sequence revealed that it contains a presumed signal peptide which is processed after Ala at position 25 from the N-terminal, and that the of the mature form is 75000. The amino acid sequences of the N-terminal and C-terminal regions of CwbA were found to be highly homologous with those of the cell wall binding domain of CwlB and the gene product, respectively. CwbA stimulated the major autolysin activity approximately threefold . These data indicate that CwbA is the modifier protein of the major autolysin reported by Herbold, D. R. & Glaser, L. (1975; 250, 1676–1682). In-frame fusion between the and genes demonstrated that is translated and expressed during the exponential growth phase.


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