Bacteriolytic activity and specificity of Achromobacter beta-lytic protease.
Achromobacter beta-lytic protease (blp), one of the bacteriolytic proteases secreted by Achromobacter lyticus, exhibited both peptidase and bacteriolytic activities at alkaline pH. The protease was strongly inhibited by 1,10-phenanthroline, and one zinc atom was detected in the molecule by ion-spray mass spectrometry. The zinc-protease specifically cleaved Gly-X bonds in peptides and possibly possessed subsites S2, S1, S1', and S2' for binding substrate [Schecter, I. and Berger, A. (1967) Biochem. Biophys. Res. Commun. 27, 157-162]. Blp lysed Staphylococcus aureus and Micrococcus luteus cells more efficiently than Achromobacter alpha-lytic protease (alp) and lysozyme, thus being responsible for the high bacteriolytic activity of A. lyticus. In the lysis of bacterial cell walls, blp hydrolyzed both the D-Ala-Gly/Ala bond at the linkage between the peptide subunit and the interpeptide and the Gly-Gly bond in the interpeptide bridge. These results indicate that blp is a highly active bacteriolytic enzyme with a broad bacteriolytic spectrum, which acts primarily by splitting the linkage between the peptide subunit and the interpeptide in the peptidoglycan.
Li
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S
Division of Protein Chemistry, Institute for Protein Research, Osaka University, Suita, Osaka, 565-0871, Japan. lishao@protein.osaka-u.ac. jp
Norioka
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S
Sakiyama
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D000421:Alcaligenes / Q000201:enzymology*; D000596:Amino Acids / Q000032:analysis; D001433:Bacteriolysis / Q000187:drug effects; D004791:Enzyme Inhibitors / Q000494:pharmacology; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis / Q000187:drug effects; D013058:Mass Spectrometry; D009994:Osmolar Concentration; D010455:Peptides / Q000378:metabolism; D010457:Peptidoglycan / Q000378:metabolism; D012697:Serine Endopeptidases / Q000378:metabolism*; D013379:Substrate Specificity
D000596:Amino Acids; D004791:Enzyme Inhibitors; D010455:Peptides; D010457:Peptidoglycan; D012697:Serine Endopeptidases; C036813:lysyl endopeptidase