Journal Article

Klebsiella phages representing a novel clade of viruses with an unknown DNA modification and biotechnologically interesting enzymes

Cava, Felipe, Drulis-Kawa, Zuzanna, Espaillat, Akbar, Briers, Yves, Kęsik-Szeloch, Agata, Lavigne, Rob, Kropinski, Andrew M., Roszniowski, Bartosz, Maciejewska, Barbara, Majkowska-Skrobek, Grazyna

2017

Overview

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.

• Publisher: Springer Berlin Heidelberg,Springer,Springer Nature B.V
• Subject: alanine / Analysis / Applied Genetics and Molecular Biotechnology / Bacteria / Bacteriophage / Bacteriophages / Bacteriophages - classification / Bacteriophages - enzymology / Bacteriophages - genetics / Bacteriophages - isolation & purification / Binding sites / Biomedical and Life Sciences / Biotechnology / Chromatography / Deoxyribonucleic acid / DNA / DNA modification / DNA sequencing / DNA, Viral - chemistry / DNA, Viral - genetics / endolysin / Endopeptidases - metabolism / Enterobacter / Enzymes / epithelial cells / Gene Order / Genes / Genetic aspects / Genomes / humans / Internet resources / Klebsiella / Klebsiella oxytoca / Klebsiella oxytoca - virology / Klebsiella pneumoniae / Klebsiella pneumoniae - virology / Klebsiella spp / Kp15virus / Laboratories / Life Sciences / Methods / Microbial Genetics and Genomics / Microbiology / Multidrug resistant organisms / multiple drug resistance / Myoviridae / Myoviridae - classification / Myoviridae - enzymology / Myoviridae - genetics / Myoviridae - isolation & purification / Nosocomial infections / nucleotide sequences / peptidoglycans / Phages / Phylogeny / Proteins / sequence analysis / Sequence Homology / Studies / Thermostable endolysin / toxicity / Transfer RNA / Virulence / Viruses