title: Enhanced staphylolytic activity of the Staphylococcus aureus bacteriophage vB_SauS-phiIPLA88 HydH5 virion-associated peptidoglycan hydrolase: fusions, deletions, and synergy with LysH5.
abstract: Virion-associated peptidoglycan hydrolases have potential as antimicrobial agents due to their ability to lyse Gram-positive bacteria on contact. In this work, our aim was to improve the lytic activity of HydH5, a virion-associated peptidoglycan hydrolase from the Staphylococcus aureus bacteriophage vB_SauS-phiIPLA88. Full-length HydH5 and two truncated derivatives containing only the CHAP (cysteine, histidine-dependent amidohydrolase/peptidase) domain exhibited high lytic activity against live S. aureus cells. In addition, three different fusion proteins were created between lysostaphin and HydH5, each of which showed higher staphylolytic activity than the parental enzyme or its deletion construct. Both parental and fusion proteins lysed S. aureus cells in zymograms and plate lysis and turbidity reduction assays. In plate lysis assays, HydH5 and its derivative fusions lysed bovine and human S. aureus strains, the methicillin-resistant S. aureus (MRSA) strain N315, and human Staphylococcus epidermidis strains. Several nonstaphylococcal bacteria were not affected. HydH5 and its derivative fusion proteins displayed antimicrobial synergy with the endolysin LysH5 in vitro, suggesting that the two enzymes have distinct cut sites and, thus, may be more efficient in combination for the elimination of staphylococcal infections.
authors.affiliation: Instituto de Productos Lácteos de Asturias (IPLA-CSIC), Asturias, Spain.
authors.forename: Lorena
authors.initials: L
authors.lastname: Rodríguez-Rubio
authors.forename: Beatriz
authors.initials: B
authors.lastname: Martínez
authors.forename: Ana
authors.initials: A
authors.lastname: Rodríguez
authors.forename: David M
authors.initials: DM
authors.lastname: Donovan
authors.forename: Pilar
authors.initials: P
authors.lastname: García
mesh_terms: D000818:Animals; D000900:Anti-Bacterial Agents / Q000627:therapeutic use; D001433:Bacteriolysis; D002417:Cattle; D002418:Cattle Diseases / Q000382:microbiology / Q000628:therapy; D004357:Drug Synergism; D010450:Endopeptidases / Q000378:metabolism; D017353:Gene Deletion; D006801:Humans; D055624:Methicillin-Resistant Staphylococcus aureus / Q000821:virology*; D008969:Molecular Sequence Data; D009238:N-Acetylmuramoyl-L-alanine Amidase / Q000235:genetics / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D013203:Staphylococcal Infections / Q000382:microbiology* / Q000628:therapy; D013204:Staphylococcus Phages / Q000201:enzymology / Q000235:genetics / Q000502:physiology; D013211:Staphylococcus aureus / Q000821:virology*; D013212:Staphylococcus epidermidis / Q000821:virology*; D014771:Virion / Q000201:enzymology
chemical_list: D000900:Anti-Bacterial Agents; D011993:Recombinant Fusion Proteins; D010450:Endopeptidases; C001143:endolysin; D009238:N-Acetylmuramoyl-L-alanine Amidase