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Staphylococcus aureus Bacteriophage 52 Endolysin Exhibits Anti-Biofilm and Broad Antibacterial Activity Against Gram-Positive Bacteria

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Abstract

Bacteriophage endolysins have been shown to hold great promise as new antibacterial agents for animal and human health in food preservation. In the present study, endolysin from Staphylococcus aureus subsp. aureus ATCC 27692-B1 bacteriophage 52 (LysSA52) was cloned, expressed, and characterized for its antimicrobial properties. Following DNA extraction from bacteriophage 52, a 1446-bp DNA fragment containing the endolysin gene (lysSA52) was obtained by PCR amplification and cloned into pET SUMO expression vector. The positive clone was validated by sequencing and open-reading frame analysis. The LysSA52 sequence shared high homology with staphylococcal phage endolysins of the SA12, SA13, and DSW2 phages and others. The cloned lysSA52 gene encoding 481 amino acids endolysin was expressed in Escherichia coli BL21 with a calculated molecular mass of 66 kDa (LysSA52). This recombinant endolysin LysSA52 exhibited lytic activity against 8 of 10 Gram-positive bacteria via agar spot-on lawn antimicrobial assay, including methicillin-resistant Staphylococcus aureus, Staphylococcus epidermidis, Staphylococcus haemolyticus, Streptococcus pneumonia, Streptococcus pyogenes, Enterococcus faecium, Enterococcus faecalis, and Bacillus atrophaeus. In addition, the 0.50 mg/mL, LysSA52 endolysins reduced about 60% of the biofilms of S. aureus and S. epidermidis established on a microtiter plate in 12 h treatment. The data from this study indicate that LysSA52 endolysin could be used as an antibacterial protein to prevent and treat infections caused by staphylococci and several other Gram-positive pathogenic bacteria irrespective of their antibiotic resistance.

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Acknowledgements

This study was supported by the Scientific Research Project Unit at Karadeniz Technical University with Grant No.TDK-2015-5340, Türkiye.

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Authors and Affiliations

  1. Department of Medical Microbiology, Faculty of Medicine, Karadeniz Technical University, Trabzon, 61080, Turkey

    Mujib Abdulkadir Abdurahman, İnci Durukan & Ali Osman Kiliç

  2. Department of Medical Biology, Faculty of Medicine, Karadeniz Technical University, Trabzon, Turkey

    Tuba Dinçer

  3. Department of Chemistry, Faculty of Arts and Sciences, Recep Tayyip Erdogan University, Rize, Turkey

    Serap Pektaş

  4. Department of Medical Services and Techniques, Patnos Vocational School, Ağrı İbrahim Çeçen University, Ağrı, Turkey

    Ersin Karataş

  5. Department of Microbial, Cellular, and Molecular Biology, Faculty of Natural Science, Addis Ababa University, Addis Ababa, Ethiopia

    Mujib Abdulkadir Abdurahman

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Contributions

MAA performed the clooning experiments, ID performed the phage experiments, TD made bioinformatic analysis, SP performed the expression experiments, EK wrote the manuscript and performed expression and purification of the protein, AOK wrote the manuscript. All authors reviewed the manuscript.

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Correspondence to Ali Osman Kiliç.

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Abdurahman, M.A., Durukan, İ., Dinçer, T. et al. Staphylococcus aureus Bacteriophage 52 Endolysin Exhibits Anti-Biofilm and Broad Antibacterial Activity Against Gram-Positive Bacteria. Protein J 42, 596–606 (2023). https://doi.org/10.1007/s10930-023-10145-1

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