title: LambdaSa2 prophage endolysin requires Cpl-7-binding domains and amidase-5 domain for antimicrobial lysis of streptococci.
abstract: Streptococcal pathogens contribute to a wide variety of human and livestock diseases. The routine use of antibiotics to battle these pathogens has produced a new class of multidrug-resistant streptococci. Thus, there is a need for new antimicrobials. Bacteriophage endolysins (peptidoglycan hydrolases) comprise one group of new antimicrobials that are reportedly refractory to resistance development. The LambdaSa2 prophage endolysin gene was recently isolated from a Group B streptococcal genome, expressed on an Escherichia coli plasmid, and shown by homology screening and biochemical analysis to harbor an amidase-5 (endopeptidase) domain, an amidase-4 (glycosidase) domain, and two Cpl-7 cell wall-binding domains. In this study, turbidity reduction and plate lysis assays indicate that this hydrolase shows strong lytic activity toward Streptococcus pyogenes, Streptococcus dysgalactiae, Streptococcus uberis, Streptococcus equi, GES, and GGS. Deletion analysis indicates that the N-terminal endopeptidase domain with both Cpl-7 domains can lyse with a higher specific activity than the full-length protein (against some strains). This dual Cpl-7 domain truncated version also shows weak lytic activity against methicillin-resistant Staphylococcus aureus (MRSA) and the coagulase negative staphylococci, Staphylococcus xylosus. The truncated constructs harboring the glycosidase domain are virtually inactive, showing only minimal activity on plate lysis assays.
authors.affiliation: Animal Biosciences and Biotechnology Lab, ANRI, ARS, USDA, Beltsville, MD 20705-2350, USA. david.donovan@ars.usda.gov
authors.forename: David M
authors.initials: DM
authors.lastname: Donovan
authors.forename: Juli
authors.initials: J
authors.lastname: Foster-Frey
mesh_terms: D000581:Amidohydrolases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D000900:Anti-Bacterial Agents / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology*; D001433:Bacteriolysis / Q000187:drug effects; D002473:Cell Wall / Q000378:metabolism; D010450:Endopeptidases / Q000737:chemistry* / Q000235:genetics / Q000494:pharmacology*; D008826:Microbial Sensitivity Tests; D039002:Prophages / Q000201:enzymology* / Q000235:genetics / Q000378:metabolism; D017434:Protein Structure, Tertiary; D017384:Sequence Deletion; D013291:Streptococcus / Q000187:drug effects / Q000821:virology; D017108:Streptococcus Phages / Q000201:enzymology*
chemical_list: D000900:Anti-Bacterial Agents; D010450:Endopeptidases; C001143:endolysin; D000581:Amidohydrolases; C019665:amidase