Isolation and characterization of hemolysin activated by reductant from Prevotella intermedia.
The hemolysin from Prevotella intermedia was partially purified from culture supernatant and then characterized. The hemolysin produced a clear beta-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.
Takada
Kazuko
K
Department of Microbiology, Nihon University School of Dentistry at Matsudo, 2-870-1 Sakaecho-nishi, Matsudo, Chiba 271-8587, Japan.
Fukatsu
Akira
A
Otake
Shigeo
S
Hirasawa
Masatomo
M
D003470:Culture Media; D006460:Hemolysin Proteins / Q000096:biosynthesis* / Q000187:drug effects / Q000302:isolation & purification*; D006358:Hot Temperature; D006863:Hydrogen-Ion Concentration; D018713:Prevotella intermedia / Q000254:growth & development / Q000378:metabolism*; D019163:Reducing Agents / Q000494:pharmacology*; D014357:Trypsin / Q000378:metabolism
D003470:Culture Media; D006460:Hemolysin Proteins; D019163:Reducing Agents; D014357:Trypsin