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Two-step purification and partial characterization of a variant of the Vibrio cholerae non-O1 hemolysin
B A McCardell1, M H Kothary, J M Madden
1Division of Virulence Assessment, Center for Food Safety and Applied Nutrition, FDA, Washington, DC 20204, USA. Bmccarde@bangate.fda.gov
Abstract:
A two-step purification method using ammonium sulfate precipitation and gel filtration was developed for the purification of a variant of the El Tor hemolysin/cytolysin from supernatant fluids of a Vibrio cholerae non-O1 human isolate (strain 2194c). The toxin displayed delayed elution from a Sephacryl gel filtration column, eluting at between two and three column volumes. The molecular mass and isoelectric point of the purified 2194c toxin were 60 kDa and 5. 3, respectively. The N-terminal amino acid sequence was ASPAPANSETNTLPHVAFYI. Purified toxin was cytolytic for Chinese hamster ovary cells and erythrocytes from several animal species.