Isolation and characterisation of KP34—a novel φKMV-like bacteriophage for Klebsiella pneumoniae

Drulis-Kawa, Zuzanna; Mackiewicz, Paweł; Kęsik-Szeloch, Agata; Maciaszczyk-Dziubińska, Ewa; Weber‐Dąbrowska, Beata; Dorotkiewicz-Jach, Agata; Augustyniak, Daria; Majkowska-Skrobek, Grażyna; Bocer, Tomasz; Empel, Joanna; Kropinski, Andrew M.

doi:10.1007/s00253-011-3149-y

Cited by 60 publications

(59 citation statements)

References 40 publications

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“…Such high homology within morphologically similar phages may indicate the presence in this part of a domain anchoring the tail fiber to the phage particle. In turn, the exhibition of a C-terminal sequence similarity to the gp57 of Klebsiella phage KP34 from Podoviridae , and the identification of common host spectrum for both phages [21,44] suggests that it is the region responsible for recognizing polysaccharides and binding to the host cell receptor. Also, a pectate lyase domain fragment, found in the sequence alignment, indicates the involvement of depoKP36 in the modification or cleavage of glycoside bonds in the capsular polysaccharide.…”

Section: Discussionmentioning

confidence: 99%

Capsule-Targeting Depolymerase, Derived from Klebsiella KP36 Phage, as a Tool for the Development of Anti-Virulent Strategy

Majkowska-Skrobek

Łątka

Berisio

et al. 2016

Viruses

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120

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The rise of antibiotic-resistant Klebsiella pneumoniae, a leading nosocomial pathogen, prompts the need for alternative therapies. We have identified and characterized a novel depolymerase enzyme encoded by Klebsiella phage KP36 (depoKP36), from the Siphoviridae family. To gain insights into the catalytic and structural features of depoKP36, we have recombinantly produced this protein of 93.4 kDa and showed that it is able to hydrolyze a crude exopolysaccharide of a K. pneumoniae host. Using in vitro and in vivo assays, we found that depoKP36 was also effective against a native capsule of clinical K. pneumoniae strains, representing the K63 type, and significantly inhibited Klebsiella-induced mortality of Galleria mellonella larvae in a time-dependent manner. DepoKP36 did not affect the antibiotic susceptibility of Klebsiella strains. The activity of this enzyme was retained in a broad range of pH values (4.0–7.0) and temperatures (up to 45 °C). Consistently, the circular dichroism (CD) spectroscopy revealed a highly stability with melting transition temperature (Tm) = 65 °C. In contrast to other phage tailspike proteins, this enzyme was susceptible to sodium dodecyl sulfate (SDS) denaturation and proteolytic cleavage. The structural studies in solution showed a trimeric arrangement with a high β-sheet content. Our findings identify depoKP36 as a suitable candidate for the development of new treatments for K. pneumoniae infections.

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Section: Discussionmentioning

confidence: 99%

Capsule-Targeting Depolymerase, Derived from Klebsiella KP36 Phage, as a Tool for the Development of Anti-Virulent Strategy

Majkowska-Skrobek

Łątka

Berisio

et al. 2016

Viruses

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120

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“…Recently, Pseudomonas fluorescens phage phi-2 has been proposed as a member of this genus (46). In addition, Klebsiella phage KP34 (13), currently assigned to the unclassified Podoviridae, also shares the typical genome architecture. In this paper we were able to extend the genus "phiKMV-like viruses" with the novel Pantoea phages LIMEzero and LIMElight.…”

Section: Resultsmentioning

confidence: 99%

“…The former has two tail fiber proteins and a tail spike protein, like phiKMV; the latter encodes a single tail fiber protein with a predicted EPS depolymerase. This EPS depolymerase showed strong homology to counterparts in Erwinia amylovora phages phi-Ea1h (27) and Era103 (56), ex- (8,13,25,34). VP93 is the only phage with two tail fiber genes (5).…”

Section: Resultsmentioning

confidence: 99%

Bacteriophages LIMElight and LIMEzero of Pantoea agglomerans, Belonging to the “phiKMV-Like Viruses”

Adriaenssens

Ceyssens

Dunon

et al. 2011

Appl Environ Microbiol

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Pantoea agglomerans is a common soil bacterium used in the biocontrol of fungi and bacteria but is also an opportunistic human pathogen. It has been described extensively in this context, but knowledge of bacteriophages infecting this species is limited. Bacteriophages LIMEzero and LIMElight of P. agglomerans are lytic phages, isolated from soil samples, belonging to the Podoviridae and are the first Pantoea phages of this family to be described. The double-stranded DNA ( Pantoea agglomerans bacteria are Gram-negative, nonsporulating, facultatively anaerobic rods that belong to the family of the Enterobacteriaceae. Synonyms of this species name are Enterobacter agglomerans, Erwinia herbicola, and Erwinia milletiae (6, 19). The genus name Pantoea is derived from the Greek word pantoios, which means "of all sorts and sources," a descriptor well reflected in the habitat, since species of the genus have been isolated from plant surfaces, seeds, water, soil, and even humans and animals (3,19,41). Pantoea agglomerans is known in plant production as a biocontrol agent. It is used in postharvest control of a number of fungi (for instance, Penicillium expansum, Rhizopus stolonifer, Monilinia laxa, and Botrytis cinerea) on a wide variety of fruits (18,44,45,54).

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“…K. pneumoniae -specific bacteriophages were isolated from sewage samples by enrichment technique described previously (Drulis-Kawa et al 2011; Kęsik-Szeloch et al 2013). Strain K. pneumoniae ATCC 700603 and clinical strain K. pneumoniae 767 ESBL(+) (Polish Collection of Microorganisms (PCM) B/F/00064) were used for amplification of KP15 (PCM F/00063) and KP27 (PCM F/00064) respectively.…”

Section: Methodsmentioning

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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Isolation and characterisation of KP34—a novel φKMV-like bacteriophage for Klebsiella pneumoniae

Cited by 60 publications

References 40 publications

Capsule-Targeting Depolymerase, Derived from Klebsiella KP36 Phage, as a Tool for the Development of Anti-Virulent Strategy

Capsule-Targeting Depolymerase, Derived from Klebsiella KP36 Phage, as a Tool for the Development of Anti-Virulent Strategy

Bacteriophages LIMElight and LIMEzero of Pantoea agglomerans, Belonging to the “phiKMV-Like Viruses”

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

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