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Characteristics and Lytic Activity of Phage-Derived Peptidoglycan Hydrolase, LysSAP8, as a Potent Alternative Biocontrol Agent for Staphylococcus aureus

Journal of Microbiology and Biotechnology 2019년 29권 12호 p.1916 ~ 1924
 ( Yu Jun-Hyeok ) - Gachon University College of BioNano Technology Department of Food Science and Biotechnology

임정아 ( Lim Jeong-A ) - Korea Food Research Institute Research Group of Consumer Safety
장현주 ( Chang Hyun-Joo ) - Korea Food Research Institute Research Group of Consumer Safety
박종현 ( Park Jong-Hyun ) - Gachon University College of BioNano Technology Department of Food Science and Biotechnology

Abstract


Outbreaks of staphylococcal food poisoning (SFP) causing serious human diseases and economic losses have been reported globally. Furthermore, the spread of Staphylococcus aureus with increased resistance to multiple antimicrobial agents has become a major concern in the food industries and medicine. Here, we isolated an endolysin LysSAP8, as one of the peptidoglycan hydrolases, derived from the bacteriophage SAP8 infecting S. aureus. This endolysin was tagged with a 6×His at the C-terminal of the target protein and purified using affinity chromatography. LysSAP8 demonstrated lytic activity against a broad spectrum of bacteria, which included a majority of the staphylococcal strains tested in this study as well as the methicillin-resistant S. aureus (MRSA); however, no such activity was observed against other gram-positive or gram-negative bacteria. Additionally, LysSAP8 could maintain bactericidal activity until 0.1 nM working concentration and after heat treatment at 37°C for 30 min. The ability of LysSAP8 to lyse cells under varying conditions of temperature (4?43°C), pH (3?9), and NaCl concentrations (0?1,000 mM), and divalent metal ions (Ca2+, Co2+, Cu2+, Mg2+, Mn2+, Hg2+, and Zn2+) was examined. At the optimized condition, LysSAP8 could disrupt approximately 3.46 log CFU/ml of the planktonic cells in their exponential phase of growth within 30 min. In this study, we have suggested that LysSAP8 could be a potent alternative as a biocontrol agent that can be used to combat MRSA.

키워드

Staphylococcus aureus; bacteriophage; endolysin; MRSA
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