Genome analysis of Cronobacter phage vB_CsaP_Ss1 reveals an endolysin with potential for biocontrol of Gram-negative bacterial pathogens

Endersen, Lorraine; Guinane, Caitriona M.; Johnston, Christopher D; Neve, Horst; Coffey, Aidan; Ross, R. Paul; McAuliffe, Olivia; O’Mahony, Jim

doi:10.1099/vir.0.068494-0

Cited by 23 publications

(13 citation statements)

References 99 publications

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“…A special interest is focussed on phage enzymes involved in the first step of viral infection responsible for bacterial envelope degradation, named depolymerases, as well as on proteins encoded by lysis cassette genes such as endolysins (Drulis-Kawa et al 2015). Although the most studies on endolysin efficacy focus on Gram-positive bacteria, it is already known that these enzymes may be successfully used against Gram-negative representatives, as well (Endersen et al 2015; Gerstmans et al 2016). The mixture or fusion with peptides with OM-disrupting properties makes these enzymes permissible though outer membrane barrier reaching the target—the murein.…”

Section: Discussionmentioning

confidence: 99%

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

Maciejewska

Roszniowski

Espaillat

et al. 2016

Appl Microbiol Biotechnol

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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.Electronic supplementary materialThe online version of this article (doi:10.1007/s00253-016-7928-3) contains supplementary material, which is available to authorized users.

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Section: Discussionmentioning

confidence: 99%

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

Maciejewska

Roszniowski

Espaillat

et al. 2016

Appl Microbiol Biotechnol

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“…Analysis of the genome sequence of bacteriophages considered for use as a biocontrol agent is essential. This is to ensure that the phage is strictly lytic and does not encode any phage lysogeny factors, virulence-related genes and/or antibiotic resistance genes ( Endersen et al, 2015 ). The complete genomic sequence analysis of bacteriophage phiC119 revealed the absence of virulence-encoding genes, potential immunoreactive allergens, and lysogeny genes.…”

Section: Discussionmentioning

confidence: 99%

Characterization of novel bacteriophage phiC119 capable of lysing multidrug-resistant Shiga toxin-producingEscherichia coliO157:H7

Amarillas

Cháidez

González‐Robles

et al. 2016

PeerJ

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BackgroundShiga toxin-producing Escherichia coli (STEC) is one of the most common and widely distributed foodborne pathogens that has been frequently implicated in gastrointestinal and urinary tract infections. Moreover, high rates of multiple antibiotic-resistant E. coli strains have been reported worldwide. Due to the emergence of antibiotic-resistant strains, bacteriophages are considered an attractive alternative to biocontrol pathogenic bacteria. Characterization is a preliminary step towards designing a phage for biocontrol.MethodsIn this study, we describe the characterization of a bacteriophage designated phiC119, which can infect and lyse several multidrug-resistant STEC strains and some Salmonella strains. The phage genome was screened to detect the stx-genes using PCR, morphological analysis, host range was determined, and genome sequencing were carried out, as well as an analysis of the cohesive ends and identification of the type of genetic material through enzymatic digestion of the genome.ResultsAnalysis of the bacteriophage particles by transmission electron microscopy showed that it had an icosahedral head and a long tail, characteristic of the family Siphoviridae. The phage exhibits broad host range against multidrug-resistant and highly virulent E. coli isolates. One-step growth experiments revealed that the phiC119 phage presented a large burst size (210 PFU/cell) and a latent period of 20 min. Based on genomic analysis, the phage contains a linear double-stranded DNA genome with a size of 47,319 bp. The phage encodes 75 putative proteins, but lysogeny and virulence genes were not found in the phiC119 genome.ConclusionThese results suggest that phage phiC119 may be a good biological control agent. However, further studies are required to ensure its control of STEC and to confirm the safety of phage use.

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“…These opportunistic pathogens are now under much scrutiny because of their ability to survive heat, desiccation, and acid stress, which poses a risk of contamination of various milk powders, herbal teas, and other dried products ( ). Enderson et al [ 129 , 130 ] expressed and purified a peptidoglycan hydrolase (LysCs4) from C. sakazakii that had the highest sequence similarity to a putative lysozyme from the temperate Cronobacter phage ES2. The purified lysozyme could degrade the peptidoglycan from both Gram-negative and Gram-positive bacteria belonging to six different genera, and could lyse outer membrane-permeabilized C. sakazakii .…”

Section: Endolysins As Food and Environmental Decontaminantsmentioning

confidence: 99%

Potential for Bacteriophage Endolysins to Supplement or Replace Antibiotics in Food Production and Clinical Care

et al. 2018

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There is growing concern about the emergence of bacterial strains showing resistance to all classes of antibiotics commonly used in human medicine. Despite the broad range of available antibiotics, bacterial resistance has been identified for every antimicrobial drug developed to date. Alarmingly, there is also an increasing prevalence of multidrug-resistant bacterial strains, rendering some patients effectively untreatable. Therefore, there is an urgent need to develop alternatives to conventional antibiotics for use in the treatment of both humans and food-producing animals. Bacteriophage-encoded lytic enzymes (endolysins), which degrade the cell wall of the bacterial host to release progeny virions, are potential alternatives to antibiotics. Preliminary studies show that endolysins can disrupt the cell wall when applied exogenously, though this has so far proven more effective in Gram-positive bacteria compared with Gram-negative bacteria. Their potential for development is furthered by the prospect of bioengineering, and aided by the modular domain structure of many endolysins, which separates the binding and catalytic activities into distinct subunits. These subunits can be rearranged to create novel, chimeric enzymes with optimized functionality. Furthermore, there is evidence that the development of resistance to these enzymes may be more difficult compared with conventional antibiotics due to their targeting of highly conserved bonds.

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Genome analysis of Cronobacter phage vB_CsaP_Ss1 reveals an endolysin with potential for biocontrol of Gram-negative bacterial pathogens

Cited by 23 publications

References 99 publications

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

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

Characterization of novel bacteriophage phiC119 capable of lysing multidrug-resistant Shiga toxin-producingEscherichia coliO157:H7

Potential for Bacteriophage Endolysins to Supplement or Replace Antibiotics in Food Production and Clinical Care

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