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

Title: Capsule-Targeting Depolymerase, Derived from Klebsiella KP36 Phage, as a Tool for the Development of Anti-Virulent Strategy
Author(s): Majkowska-Skrobek, G
Latka, A
Berisio, R
Maciejewska, B
Squeglia, F
Romano, M
Lavigne, R
Struve, C
Drulis-Kawa, Z
Item Type: Journal Article
Abstract: 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.
Publication Date: 1-Dec-2016
Date of Acceptance: 23-Nov-2016
URI: http://hdl.handle.net/10044/1/53039
DOI: https://dx.doi.org/10.3390/v8120324
ISSN: 1999-4915
Publisher: MDPI AG
Journal / Book Title: Viruses
Volume: 8
Issue: 12
Copyright Statement: © 2016 by the authors; licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC-BY) license (http://creativecommons.org/licenses/by/4.0/).
Keywords: Science & Technology
Life Sciences & Biomedicine
Virology
bacteriophage
Klebsiella sp
capsule
polysaccharide depolymerase
ESCHERICHIA-COLI PHAGE
TAIL SPIKE
CRYSTAL-STRUCTURE
HK620 TAILSPIKE
BETA-HELIX
BACTERIOPHAGE
PNEUMONIAE
PROTEIN
POLYSACCHARIDE
INFECTION
Klebsiella sp.
bacteriophage
capsule
polysaccharide depolymerase
Animals
Anti-Bacterial Agents
Bacterial Capsules
Bacteriophages
Circular Dichroism
Cloning, Molecular
Disease Models, Animal
Enzyme Stability
Gene Expression
Glycoside Hydrolases
Hydrogen-Ion Concentration
Klebsiella Infections
Klebsiella pneumoniae
Lepidoptera
Molecular Weight
Protein Conformation
Temperature
Treatment Outcome
Animals
Lepidoptera
Klebsiella pneumoniae
Bacteriophages
Klebsiella Infections
Disease Models, Animal
Glycoside Hydrolases
Bacterial Capsules
Anti-Bacterial Agents
Treatment Outcome
Circular Dichroism
Cloning, Molecular
Enzyme Stability
Temperature
Gene Expression
Protein Conformation
Hydrogen-Ion Concentration
Molecular Weight
Science & Technology
Life Sciences & Biomedicine
Virology
bacteriophage
Klebsiella sp
capsule
polysaccharide depolymerase
ESCHERICHIA-COLI PHAGE
TAIL SPIKE
CRYSTAL-STRUCTURE
HK620 TAILSPIKE
BETA-HELIX
BACTERIOPHAGE
PNEUMONIAE
PROTEIN
POLYSACCHARIDE
INFECTION
Publication Status: Published
Article Number: 324
Appears in Collections:Faculty of Natural Sciences



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