purification of epsilon toxin from vaccinal strain of clostridium perfringens type D using ion exchange chromatography and gel filtration

Clostridium perfringens type ِD strains cause severe enteroxemia in sheep، calves، goats، pigs and chickens. epsilon-toxin is introduced to be the essential virulence factor of this microorganism. In the present study، a new method was established for the purification of epsilon toxin from culture supernatant fluid of Clostridium perfringens type D vaccinal strain. The three steps of the purification scheme involved ammonium sulfate precipitation، cation exchange chromatography (CM- Sepharose) and gel filtration (Sephadex G-100). epsilon toxin was purified about 138. 9-fold from the Sephadex G-100 column in terms of the toxin lethality. The molecular weight of the epsilon toxin determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis was approximately 34kD. MDCK exposed to epsilon toxin showed a cell border retraction، cytoplasmic blebbing، cell shrinkage and cell rounding. In conclusion، this new method for purification of epsilon toxin of clostridium perfringens type D is reliable and the purified epsilon toxin has high activity.