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Enzymology and Protein Engineering

Lysin LysMK34 of Acinetobacter baumannii Bacteriophage PMK34 Has a Turgor Pressure-Dependent Intrinsic Antibacterial Activity and Reverts Colistin Resistance

Karim Abdelkader, Diana Gutiérrez, Dennis Grimon, Patricia Ruas-Madiedo, Cédric Lood, Rob Lavigne, Amal Safaan, Ahmed S. Khairalla, Yasser Gaber, Tarek Dishisha, Yves Briers
Harold L. Drake, Editor
Karim Abdelkader
aDepartment of Biotechnology, Ghent University, Ghent, Belgium
bDepartment of Microbiology and Immunology, Faculty of Pharmacy, Beni-Suef University, Beni-Suef, Egypt
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Diana Gutiérrez
aDepartment of Biotechnology, Ghent University, Ghent, Belgium
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Dennis Grimon
aDepartment of Biotechnology, Ghent University, Ghent, Belgium
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Patricia Ruas-Madiedo
cDairy Research Institute of Asturias, Spanish National Research Council (IPLA-CSIC), Villaviciosa, Asturias, Spain
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Cédric Lood
dDepartment of Biosystems, KU Leuven, Leuven, Belgium
eDepartment of Microbial and Molecular Systems, Centre of Microbial and Plant Genetics, Laboratory of Computational Systems Biology, KU Leuven, Leuven, Belgium
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Rob Lavigne
dDepartment of Biosystems, KU Leuven, Leuven, Belgium
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Amal Safaan
fDepartment of Microbiology and Immunology, Faculty of Pharmacy, Menoufia University, Shebin ElKoum, Egypt
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Ahmed S. Khairalla
bDepartment of Microbiology and Immunology, Faculty of Pharmacy, Beni-Suef University, Beni-Suef, Egypt
hDepartment of Biology, University of Regina, Regina, Saskatchewan, Canada
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Yasser Gaber
bDepartment of Microbiology and Immunology, Faculty of Pharmacy, Beni-Suef University, Beni-Suef, Egypt
gDepartment of Pharmaceutics and Pharmaceutical Technology, College of Pharmacy, Mutah University, Karak, Jordan
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Tarek Dishisha
bDepartment of Microbiology and Immunology, Faculty of Pharmacy, Beni-Suef University, Beni-Suef, Egypt
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Yves Briers
aDepartment of Biotechnology, Ghent University, Ghent, Belgium
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Harold L. Drake
University of Bayreuth
Roles: Editor
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DOI: 10.1128/AEM.01311-20
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ABSTRACT

The prevalence of extensively and pandrug-resistant strains of Acinetobacter baumannii leaves little or no therapeutic options for treatment for this bacterial pathogen. Bacteriophages and their lysins represent attractive alternative antibacterial strategies in this regard. We used the extensively drug-resistant A. baumannii strain MK34 to isolate the bacteriophage PMK34 (vB_AbaP_PMK34). This phage shows fast adsorption and lacks virulence genes; nonetheless, its narrow host spectrum based on capsule recognition limits broad application. PMK34 is a Fri1virus member of the Autographiviridae and has a 41.8-kb genome (50 open reading frames), encoding an endolysin (LysMK34) with potent muralytic activity (1,499.9 ± 131 U/μM), a typical mesophilic thermal stability up to 55°C, and a broad pH activity range (4 to 10). LysMK34 has an intrinsic antibacterial activity up to 4.8 and 2.4 log units for A. baumannii and Pseudomonas aeruginosa strains, respectively, but only when a high turgor pressure is present. The addition of 0.5 mM EDTA or application of an osmotic shock after treatment can compensate for the lack of a high turgor pressure. The combination of LysMK34 and colistin results in up to 32-fold reduction of the MIC of colistin, and colistin-resistant strains are resensitized in both Mueller-Hinton broth and 50% human serum. As such, LysMK34 may be used to safeguard the applicability of colistin as a last-resort antibiotic.

IMPORTANCE A. baumannii is one of the most challenging pathogens for which development of new and effective antimicrobials is urgently needed. Colistin is a last-resort antibiotic, and even colistin-resistant A. baumannii strains exist. Here, we present a lysin that sensitizes A. baumannii for colistin and can revert colistin resistance to colistin susceptibility. The lysin also shows a strong, turgor pressure-dependent intrinsic antibacterial activity, providing new insights in the mode of action of lysins with intrinsic activity against Gram-negative bacteria.

FOOTNOTES

    • Received 2 June 2020.
    • Accepted 16 July 2020.
    • Accepted manuscript posted online 24 July 2020.
  • Supplemental material is available online only.

  • Copyright © 2020 American Society for Microbiology.

All Rights Reserved.

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Lysin LysMK34 of Acinetobacter baumannii Bacteriophage PMK34 Has a Turgor Pressure-Dependent Intrinsic Antibacterial Activity and Reverts Colistin Resistance
Karim Abdelkader, Diana Gutiérrez, Dennis Grimon, Patricia Ruas-Madiedo, Cédric Lood, Rob Lavigne, Amal Safaan, Ahmed S. Khairalla, Yasser Gaber, Tarek Dishisha, Yves Briers
Applied and Environmental Microbiology Sep 2020, 86 (19) e01311-20; DOI: 10.1128/AEM.01311-20

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Lysin LysMK34 of Acinetobacter baumannii Bacteriophage PMK34 Has a Turgor Pressure-Dependent Intrinsic Antibacterial Activity and Reverts Colistin Resistance
Karim Abdelkader, Diana Gutiérrez, Dennis Grimon, Patricia Ruas-Madiedo, Cédric Lood, Rob Lavigne, Amal Safaan, Ahmed S. Khairalla, Yasser Gaber, Tarek Dishisha, Yves Briers
Applied and Environmental Microbiology Sep 2020, 86 (19) e01311-20; DOI: 10.1128/AEM.01311-20
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KEYWORDS

Acinetobacter baumannii
antibiotic resistance
bacteriophage
colistin
endolysin

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