title: Bacillus amyloliquefaciens phage endolysin can enhance permeability of Pseudomonas aeruginosa outer membrane and induce cell lysis.
abstract: To determine the function of the C-terminal region of Bacillus amyloliquefaciens phage endolysin on Pseudomonas aeruginosa lysis, the permeabilization of the outer membrane of P. aeruginosa was analyzed. Glu-15 to His (E15H) and Thr-32 to Glu (T32E) substitutions were introduced into the Bacillus phage endolysin. Neither E15H nor T32E substitution induced enzymatic and antibacterial activities. These two, Glu-15 and Thr-32, were considered to be the active center of the enzyme. The addition of purified E15H and T32E proteins to P. aeruginosa cells induced the release of periplasmic beta-lactamase from the cells, indicating that both proteins enhance permeabilization of the outer membrane. However, the addition of E15H and T32E proteins to P. aeruginosa cells did not induce the release of cytoplasmic ATP from the cells. These results indicate that the antibacterial activity of the endolysin requires both the C-terminal enhancement of the permeabilization of the P. aeruginosa outer membrane and N-terminal enzymatic activity.
authors.affiliation: Department of Bioengineering, Tokyo Institute of Technology, 4259 Nagatsuta, Midori, 226-8501Yokohama, Japan.
authors.forename: Y
authors.initials: Y
authors.lastname: Orito
authors.forename: M
authors.initials: M
authors.lastname: Morita
authors.forename: K
authors.initials: K
authors.lastname: Hori
authors.forename: H
authors.initials: H
authors.lastname: Unno
authors.forename: Y
authors.initials: Y
authors.lastname: Tanji
mesh_terms: D000900:Anti-Bacterial Agents; D001407:Bacillus / Q000235:genetics / Q000821:virology*; D017103:Bacillus Phages / Q000737:chemistry* / Q000235:genetics; D001425:Bacterial Outer Membrane Proteins / Q000378:metabolism; D002463:Cell Membrane Permeability / Q000187:drug effects*; D010450:Endopeptidases / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology*; D011550:Pseudomonas aeruginosa / Q000187:drug effects* / Q000502:physiology
chemical_list: D000900:Anti-Bacterial Agents; D001425:Bacterial Outer Membrane Proteins; D010450:Endopeptidases; C001143:endolysin