title: Klebsiella phages representing a novel clade of viruses with an unknown DNA modification and biotechnologically interesting enzymes.
abstract: Lytic bacteriophages and phage-encoded endolysins (peptidoglycan hydrolases) provide a source for the development of novel antimicrobial strategies. In the present study, we focus on the closely related (96 % DNA sequence identity) environmental myoviruses vB_KpnM_KP15 (KP15) and vB_KpnM_KP27 (KP27) infecting multidrug-resistant Klebsiella pneumoniae and Klebsiella oxytoca strains. Their genome organisation and evolutionary relationship are compared to Enterobacter phage phiEap-3 and Klebsiella phages Matisse and Miro. Due to the shared and distinct evolutionary history of these phages, we propose to create a new phage genus "Kp15virus" within the Tevenvirinae subfamily. In silico genome analysis reveals two unique putative homing endonucleases of KP27 phage, probably involved in unrevealed mechanism of DNA modification and resistance to restriction digestion, resulting in a broader host spectrum. Additionally, we identified in KP15 and KP27 a complete set of lysis genes, containing holin, antiholin, spanin and endolysin. By turbidimetric assays on permeabilized Gram-negative strains, we verified the ability of the KP27 endolysin to destroy the bacterial peptidoglycan. We confirmed high stability, absence of toxicity on a human epithelial cell line and the enzymatic specificity of endolysin, which was found to possess endopeptidase activity, cleaving the peptide stem between L-alanine and D-glutamic acid.
authors.affiliation: Institute of Genetics and Microbiology, University of Wroclaw, S. Przybyszewskiego 63/77, 51-148, Wrocław, Poland.
authors.forename: Barbara
authors.initials: B
authors.lastname: Maciejewska
authors.affiliation: Institute of Genetics and Microbiology, University of Wroclaw, S. Przybyszewskiego 63/77, 51-148, Wrocław, Poland.
authors.forename: Bartosz
authors.initials: B
authors.lastname: Roszniowski
authors.affiliation: Laboratory for Molecular Infection Medicine Sweden, Molecular Biology Department, Umeå University, SE-901 87, Umeå, Sweden.
authors.forename: Akbar
authors.initials: A
authors.lastname: Espaillat
authors.affiliation: Institute of Genetics and Microbiology, University of Wroclaw, S. Przybyszewskiego 63/77, 51-148, Wrocław, Poland.
authors.forename: Agata
authors.initials: A
authors.lastname: Kęsik-Szeloch
authors.affiliation: Institute of Genetics and Microbiology, University of Wroclaw, S. Przybyszewskiego 63/77, 51-148, Wrocław, Poland.
authors.forename: Grazyna
authors.initials: G
authors.lastname: Majkowska-Skrobek
authors.affiliation: Departments of Food Science, Molecular and Cellular Biology, and Pathobiology, University of Guelph, 50 Stone Road East, Guelph, ON, N1G 2W1, Canada.
authors.forename: Andrew M
authors.initials: AM
authors.lastname: Kropinski
authors.affiliation: Department Applied Biosciences, Ghent University, Valentin Vaerwyckweg 1, 9000, Ghent, Belgium.
authors.forename: Yves
authors.initials: Y
authors.lastname: Briers
authors.affiliation: Laboratory for Molecular Infection Medicine Sweden, Molecular Biology Department, Umeå University, SE-901 87, Umeå, Sweden.
authors.forename: Felipe
authors.initials: F
authors.lastname: Cava
authors.affiliation: Laboratory of Gene Technology, KULeuven, Leuven Kasteelpark Arenberg 21 Box 2462, 3001, Leuven, Belgium.
authors.forename: Rob
authors.initials: R
authors.lastname: Lavigne
authors.affiliation: Institute of Genetics and Microbiology, University of Wroclaw, S. Przybyszewskiego 63/77, 51-148, Wrocław, Poland. zuzanna.drulis-kawa@uwr.edu.pl.
authors.forename: Zuzanna
authors.initials: Z
authors.lastname: Drulis-Kawa
mesh_terms: D001435:Bacteriophages / Q000145:classification / Q000201:enzymology* / Q000235:genetics / Q000302:isolation & purification*; D004279:DNA, Viral / Q000737:chemistry* / Q000235:genetics; D010450:Endopeptidases / Q000378:metabolism*; D023061:Gene Order; D041121:Klebsiella oxytoca / Q000821:virology*; D007711:Klebsiella pneumoniae / Q000821:virology*; D017900:Myoviridae / Q000145:classification / Q000201:enzymology / Q000235:genetics / Q000302:isolation & purification; D010802:Phylogeny; D017385:Sequence Homology
chemical_list: D004279:DNA, Viral; D010450:Endopeptidases; C001143:endolysin
keywords: Bacteriophage; DNA modification; Klebsiella spp.; Kp15virus; Thermostable endolysin