Bacillus subtilis CwlP of the SP-β Prophage Has Two Novel Peptidoglycan Hydrolase Domains, Muramidase and Cross-linkage Digesting dd-Endopeptidase

• Published in: The Journal of biological chemistry Vol. 285; no. 53; pp. 41232 - 41243
• Main Authors: Sudiarta, I Putu, Fukushima, Tatsuya, Sekiguchi, Junichi
• Format: Journal Article
• Language: English
• Published: 9650 Rockville Pike, Bethesda, MD 20814, U.S.A Elsevier Inc 31.12.2010; American Society for Biochemistry and Molecular Biology
• Subjects: Bacillus; Bacillus subtilis; Bacteria; Bacteriophage; Cell Surface Enzymes; Cell Surface Protein; Cell Wall; Enzymes; Enzymology; Escherichia coli; Gram-positive Bacteria; Peptidoglycan; Staphylococcus aureus; Streptococcus thermophilus; Cell Wall; Enzymes; Peptidoglycan; Cell Surface Protein; Bacillus; Bacteria; Bacteriophage; Cell Surface Enzymes; Gram-positive Bacteria
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1074/jbc.M110.156273

For bacteria and bacteriophages, cell wall digestion by hydrolases is a very important event. We investigated one of the proteins involved in cell wall digestion, the yomI gene product (renamed CwlP). The gene is located in the SP-β prophage region of the Bacillus subtilis chromosome. Inspection of the Pfam database indicates that CwlP contains soluble lytic transglycosylase (SLT) and peptidase M23 domains, which are similar to Escherichia coli lytic transglycosylase Slt70, and the Staphylococcus aureus Gly-Gly endopeptidase LytM, respectively. The SLT domain of CwlP exhibits hydrolytic activity toward the B. subtilis cell wall; however, reverse phase (RP)-HPLC and mass spectrometry revealed that the CwlP-SLT domain has only muramidase activity. In addition, the peptidase M23 domain of CwlP exhibited hydrolytic activity and could cleave d-Ala-diaminopimelic acid cross-linkage, a property associated with dd-endopeptidases. Remarkably, the M23 domain of CwlP possessed a unique Zn2+-independent endopeptidase activity; this contrasts with all other characterized M23 peptidases (and enzymes similar to CwlP), which are Zn2+ dependent. Both domains of CwlP could hydrolyze the peptidoglycan and cell wall of B. subtilis. However, the M23 domain digested neither the peptidoglycans nor the cell walls of S. aureus or Streptococcus thermophilus. The effect of defined point mutations in conserved amino acid residues of CwlP is also determined.