Muralytic activity and modular structure of the endolysins of Pseudomonas aeruginosa bacteriophages φKZ and EL

• Published in: Molecular microbiology Vol. 65; no. 5; pp. 1334 - 1344
• Main Authors: Briers, Yves, Volckaert, Guido, Cornelissen, Anneleen, Lagaert, Stijn, Michiels, Chris W., Hertveldt, Kirsten, Lavigne, Rob
• Format: Journal Article
• Language: English
• Published: Oxford, UK Blackwell Publishing Ltd 01.09.2007; Blackwell Science
• Subjects: Bacillus; Bacillus subtilis; Bacteriology; Biological and medical sciences; Clostridium; Fundamental and applied biological sciences. Psychology; Microbiology; Miscellaneous; Phage hkz; Pseudomonas aeruginosa; Pseudomonadales; Bacteria; Pseudomonadaceae; Pseudomonas aeruginosa
• ISSN: 0950-382X; 1365-2958
• DOI: 10.1111/j.1365-2958.2007.05870.x

Summary Pseudomonas aeruginosa bacteriophage endolysins KZ144 (phage φKZ) and EL188 (phage EL) are highly lytic peptidoglycan hydrolases (210 000 and 390 000 units mg−1), active on a broad range of outer membrane‐permeabilized Gram‐negative species. Site‐directed mutagenesis indicates E115 (KZ144) and E155 (EL188) as their respective essential catalytic residues. Remarkably, both endolysins have a modular structure consisting of an N‐terminal substrate‐binding domain and a predicted C‐terminal catalytic module, a property previously only demonstrated in endolysins originating from phages infecting Gram‐positives and only in an inverse arrangement. Both binding domains contain conserved repeat sequences, consistent with those of some peptidoglycan hydrolases of Gram‐positive bacteria. Fusions of these domains with green fluorescent protein immediately label all outer membrane‐permeabilized Gram‐negative bacteria tested, isolated P. aeruginosa peptidoglycan and N‐acetylated Bacillus subtilis peptidoglycan, demonstrating the broad range of peptidoglycan‐binding capacity by these domains. Specifically, A1 chemotype peptidoglycan and fully N‐acetylated glucosamine units are essential for binding. Both KZ144 and EL188 appear to be a natural chimeric enzyme, originating from a recombination of a cell wall‐binding domain encoded by a Bacillus or Clostridium species and a catalytic domain of an unknown ancestor.