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KMID : 0364820160520030260
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Korean Journal of Microbiology
2016 Volume.52 No. 3 p.260 ~ p.268
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Cellular responses and proteomic analysis of hemolytic Bacillus cereus MH-2 exposed to epigallocatechin gallate (EGCG)
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Kim Dong-Mim
Park Sang-Kook
Oh Kye-Heon
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Abstract
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The aim of this work was to investigate the cellular responses and proteomic analysis of Bacillus cereus MH-2 exposed to
EGCG. Strain MH-2 was isolated from commercial Ssamjang and has the hemolytic activity. Survival of the MH-2 strain with time in the presence of different concentrations of EGCG under sublethal conditions was monitored. The amount of alginate from MH-2 strain decreased depending on the increasing concentrations of EGCG and increased depending on the exposure time at any particular EGCG concentration. Analysis of SDS-PAGE and Western blot using anti-DnaK and anti-GroEL revealed that two stress shock proteins, 70 kDa DnaK and 60 kDa GroEL were found to decrease in proportion to the EGCG concentration in exponentially growing cultures. Scanning electron microscopic analysis demonstrated the presence of protrusions and fused rod forms on the cells treated with EGCG. 2-DE of soluble protein fractions from MH-2 cultures showed 20 protein spots changed by EGCG exposure. These proteins involved in enterotoxins (hemolysin BL lytic component L1 and hemolysin BL-binding protein), chaperons (DnaK and GroEL), cell defense (peptidase
M4 family proteins), and various biosynthesis and energy metabolism were identified by petide mass fingerprinting using MALDI-TOF. These results provide clues for understanding the mechanism of EGCG-induced stress and cytotoxicity on B. cereus MH-2.
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KEYWORD
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Bacillus cereus MH-2, EGCG, epigallocatechin gallate, hemolysis
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