EC300: a phage-based, bacteriolysin-like protein with enhanced antibacterial activity against Enterococcus faecalis.
Bacteriophage lytic enzymes, either endolysins or virion-associated lysins, have been receiving considerable attention as potential antibacterial agents, particularly for the combat of antibiotic-resistant Gram-positive pathogens. A conclusion that easily emerges from the careful analysis of a great number of reports on the field is that the activity of phage lytic enzymes is rarely studied in conditions that support robust growth of the target bacteria. Here, we report the construction and study of a chimerical lysin, EC300, which was designed to target and kill Enterococcus faecalis in conditions supporting vigorous bacterial growth. EC300 resulted from the fusion of a predicted M23 endopeptidase domain of a virion-associated lysin to the putative cell wall binding domain of a previously characterized amidase endolysin, both produced by the E. faecalis phage F170/08. This bacteriolysin-like protein exhibited a clear enhanced lytic activity over the parental endolysin when both were assayed in a rich bacterial growth medium. We demonstrate the killing efficacy of EC300 against growing cells of a panel of typed E. faecalis clinical strains with high level of antibiotic resistance. The possible reasons for the marked difference between the lytic performance of EC300 and that of the amidase are discussed.
Technophage, SA, Av. Professor Egas Moniz, Ed. Egas Moniz, piso 2, 1649-028, Lisboa, Portugal.
Daniela
D
Proença
Clara
C
Leandro
Miguel
M
Garcia
Madalena
M
Pimentel
Carlos
C
São-José
D000900:Anti-Bacterial Agents / Q000302:isolation & purification / Q000378:metabolism / Q000494:pharmacology*; D001435:Bacteriophages / Q000201:enzymology* / Q000235:genetics; D013293:Enterococcus faecalis / Q000187:drug effects* / Q000254:growth & development; D016908:Gram-Positive Bacterial Infections / Q000382:microbiology; D006801:Humans; D009088:Mucoproteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000494:pharmacology*; D015202:Protein Engineering; D014764:Viral Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism / Q000494:pharmacology*
D000900:Anti-Bacterial Agents; D009088:Mucoproteins; D014764:Viral Proteins; C010393:lysin, gastropoda