title: The phage lytic proteins from the Staphylococcus aureus bacteriophage vB_SauS-phiIPLA88 display multiple active catalytic domains and do not trigger staphylococcal resistance.
abstract: The increase in antibiotic resistance world-wide revitalized the interest in the use of phage lysins to combat pathogenic bacteria. In this work, we analyzed the specific cleavage sites on the staphylococcal peptidoglycan produced by three phage lytic proteins. The investigated cell wall lytic enzymes were the endolysin LysH5 derived from the S. aureus bacteriophage vB_SauS-phi-IPLA88 (phi-IPLA88) and two fusion proteins between lysostaphin and the virion-associated peptidoglycan hydrolase HydH5 (HydH5SH3b and HydH5Lyso). We determined that all catalytic domains present in these proteins were active. Additionally, we tested for the emergence of resistant Staphylococcus aureus to any of the three phage lytic proteins constructs. Resistant S. aureus could not be identified after 10 cycles of bacterial exposure to phage lytic proteins either in liquid or plate cultures. However, a quick increase in lysostaphin resistance (up to 1000-fold in liquid culture) was observed. The lack of resistant development supports the use of phage lytic proteins as future therapeutics to treat staphylococcal infections.
authors.affiliation: DairySafe Group, Department of Technology and Biotechnology of Dairy Products, Instituto de Productos Lácteos de Asturias-Consejo Superior de Investigaciones Científicas, Villaviciosa, 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: Friedrich
authors.initials: F
authors.lastname: Götz
authors.forename: Pilar
authors.initials: P
authors.lastname: García
mesh_terms: D000595:Amino Acid Sequence; D020134:Catalytic Domain; D024881:Drug Resistance, Bacterial; D010450:Endopeptidases / Q000737:chemistry* / Q000378:metabolism; D008248:Lysostaphin / Q000737:chemistry* / Q000378:metabolism; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D009238:N-Acetylmuramoyl-L-alanine Amidase / Q000737:chemistry* / Q000378:metabolism; D010457:Peptidoglycan / Q000737:chemistry / Q000378:metabolism; D013204:Staphylococcus Phages / Q000378:metabolism*; D013211:Staphylococcus aureus / Q000821:virology*
chemical_list: D010457:Peptidoglycan; D010450:Endopeptidases; D008248:Lysostaphin; C001143:endolysin; D009238:N-Acetylmuramoyl-L-alanine Amidase