Open AccessIsolation and characterization of hemolysin activated by reductant from Prevotella intermedia
Author(s) -
Takada Kazuko,
Fukatsu Akira,
Otake Shigeo,
Hirasawa Masatomo
Publication year - 2003
Publication title -
fems immunology & medical microbiology
Language(s) - English
Resource type - Journals
eISSN - 1574-695X
pISSN - 0928-8244
DOI - 10.1111/j.1574-695x.2003.tb00647.x
Subject(s) - hemolysin , dithiothreitol , trypsin , biology , hemolysis , biochemistry , polyethylene glycol , enterobacter aerogenes , glutathione , agar plate , microbiology and biotechnology , chemistry , chromatography , enzyme , bacteria , escherichia coli , genetics , virulence , gene , immunology
The hemolysin from Prevotella intermedia was partially purified from culture supernatant and then characterized. The hemolysin produced a clear β‐hemolytic zone on a blood agar plate. Hemolytic activity was 2.5‐fold greater in culture supernatant compared to that cell‐associated. The isolation and purification procedure involved ammonium sulfate and polyethylene glycol precipitations and ion‐exchange chromatographies on DEAE‐Sephacel and CM‐Sepharose. The activity of this hemolysin was stimulated by reductants such as cysteine, dithiothreitol, glutathione etc., and was lost upon oxidation. Trypsin or heat treatment resulted in complete inhibition of hemolytic activity. Ca 2+ , Mg 2+ and EDTA did not affect the activity. The optimal pH of this hemolysin was 7.5.