Isolation and Characterization of a Novel Lytic Bacteriophage against the K2 Capsule-Expressing Hypervirulent Klebsiella pneumoniae Strain 52145, and Identification of Its Functional Depolymerase.

Botond Zsombor Pertics,Tamás Kovács,Alysia Cox,Adrienn Nyúl,György Schneider,Nóra Szamek

doi:10.3390/microorganisms9030650

Botond Zsombor Pertics, Tamás Kovács + Show 4 more

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https://doi.org/10.3390/microorganisms9030650

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Abstract

Klebsiella pneumoniae is among the leading bacteria that cause nosocomial infections. The capsule of this Gram-negative bacterium is a dominant virulence factor, with a prominent role in defense and biofilm formation. Bacteriophages, which are specific for one bacterial strain and its capsule type, can evoke the lysis of bacterial cells, aided by polysaccharide depolymerase enzymes. In this study, we isolated and characterized a bacteriophage against the nosocomial K. pneumoniae 52145 strain with K2 capsular serotype. The phage showed a narrow host range and stable lytic activity, even when exposed to different temperatures or detergents. Preventive effect of the phage in a nasal colonization model was investigated in vivo. Phlyogenetic analysis showed that the newly isolated Klebsiella phage B1 belongs to the Webervirus genus in Drexlerviridae family. We identified the location of the capsule depolymerase gene of the new phage, which was amplified, cloned, expressed, and purified. The efficacy of the recombinant B1dep depolymerase was tested by spotting on K. pneumoniae strains and it was confirmed that the extract lowers the thickness of the bacterium lawn as it degrades the protective capsule on bacterial cells. As K. pneumoniae strains possessing the K2 serotype have epidemiological importance, the B1 phage and its depolymerase are promising candidates for use as possible antimicrobial agents.

Highlights

Klebsiella pneumoniae is a Gram-negative, encapsulated bacterium and one of the most important opportunistic nosocomial pathogens [1]
K. pneumoniae is a member of the ESKAPE (Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa and Enterobacter spp.) group of microorganisms [6] as isolates are frequently resistant to multiple antibiotics
Bacteria were routinely grown on lysogeny broth agar (LBA) plates at 37 ◦ C or liquid lysogeny broth (LB) medium (37 ◦ C at 125 rpm)

Summary

Introduction

Klebsiella pneumoniae is a Gram-negative, encapsulated bacterium and one of the most important opportunistic nosocomial pathogens [1]. It is ubiquitous in the environment [2]. Colonizes the human skin and mucosal surfaces including the gastrointestinal tract and oropharynx, where it can be a source of serious infection of the urinary and respiratory tracts, surgical sites, and catheter entry points. These infections can progress to potentially life-threatening conditions and septicemia [1,3], most often in hospitalized and immune-compromised patients [4,5]. K. pneumoniae is a member of the ESKAPE (Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa and Enterobacter spp.) group of microorganisms [6] as isolates are frequently resistant to multiple antibiotics. 4.0/).

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