caa a22 4500 60550069X CHVBK 20210128100557.0 cr unu---uuuuu 210128e20150601xx s 000 0 eng 10.1007/s00253-015-6483-7 doi (NATIONALLICENCE)springer-10.1007/s00253-015-6483-7 EC300: a phage-based, bacteriolysin-like protein with enhanced antibacterial activity against Enterococcus faecalis [Elektronische Daten] [Daniela Proença, Clara Leandro, Miguel Garcia, Madalena Pimentel, Carlos São-José] 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. Springer-Verlag Berlin Heidelberg, 2015 Virion-associated lysin nationallicence Endolysin nationallicence Chimerical enzyme nationallicence Peptidoglycan hydrolase nationallicence Enterococcus faecalis nationallicence Proença Daniela Technophage, SA, Av. Professor Egas Moniz, Ed. Egas Moniz, piso 2, 1649-028, Lisboa, Portugal aut Leandro Clara Technophage, SA, Av. Professor Egas Moniz, Ed. Egas Moniz, piso 2, 1649-028, Lisboa, Portugal aut Garcia Miguel Technophage, SA, Av. Professor Egas Moniz, Ed. Egas Moniz, piso 2, 1649-028, Lisboa, Portugal aut Pimentel Madalena Centro de Patogénese Molecular, Unidade dos Retrovírus e Infecções Associadas (CPM-URIA), Faculdade de Farmácia da Universidade de Lisboa, Av. Prof. Gama Pinto, 1649-003, Lisboa, Portugal aut São-José Carlos Centro de Patogénese Molecular, Unidade dos Retrovírus e Infecções Associadas (CPM-URIA), Faculdade de Farmácia da Universidade de Lisboa, Av. Prof. Gama Pinto, 1649-003, Lisboa, Portugal aut Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/12(2015-06-01), 5137-5149 0175-7598 99:12<5137 2015 99 253 https://doi.org/10.1007/s00253-015-6483-7 text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s00253-015-6483-7 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Proença Daniela Technophage, SA, Av. Professor Egas Moniz, Ed. Egas Moniz, piso 2, 1649-028, Lisboa, Portugal aut NATIONALLICENCE 700 1- Leandro Clara Technophage, SA, Av. Professor Egas Moniz, Ed. Egas Moniz, piso 2, 1649-028, Lisboa, Portugal aut NATIONALLICENCE 700 1- Garcia Miguel Technophage, SA, Av. Professor Egas Moniz, Ed. Egas Moniz, piso 2, 1649-028, Lisboa, Portugal aut NATIONALLICENCE 700 1- Pimentel Madalena Centro de Patogénese Molecular, Unidade dos Retrovírus e Infecções Associadas (CPM-URIA), Faculdade de Farmácia da Universidade de Lisboa, Av. Prof. Gama Pinto, 1649-003, Lisboa, Portugal aut NATIONALLICENCE 700 1- São-José Carlos Centro de Patogénese Molecular, Unidade dos Retrovírus e Infecções Associadas (CPM-URIA), Faculdade de Farmácia da Universidade de Lisboa, Av. Prof. Gama Pinto, 1649-003, Lisboa, Portugal aut NATIONALLICENCE 773 0- Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/12(2015-06-01), 5137-5149 0175-7598 99:12<5137 2015 99 253