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KMID : 0613820020120040490
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Journal of Life Science
2002 Volume.12 No. 4 p.490 ~ p.495
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Purification and characterization of biochemical properties of hemolysin from Vibrio fluvialis
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Lee Jong-Hee
Han Jeong-Hyun
Ahn Sun-Hee
Kim Sun-Hui
Lee Eun-Mi
Kong In-Soo
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Abstract
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Hemolysin (VFH) of V. fluvialis, which is a pathogenic bacteria, causing watery diarrhea with vomiting, abdominal croup, was purified. V. fluvialis was cultivated in BHI medium and the culture supernatant was precipitated by ammonium sulfate. The protein was purified by chromatographies on columns of DEAE-cellulose and Mono-Q. Molecular weight of the purified VFH was estimated as 79kDa by SDS-PAGE. The optimal temperature for a maximum hemolytic activity was at around 35℃ and the activity was decreased at 40℃. Cytotoxicity of VFH was also investigated using RTG-2 cell line. LDH assay study showed that 50㎍/ml of VFH release 80% of total cellular LDH (lactate dehydrogenase) from RTG-2 cell and microscopic observation also showed the morphological change of cell.
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KEYWORD
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Vibrio fluvialis, Hemolysin, VFH, Cytotoxicity, RTG-2 cell
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