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

Zayerzadeh, Ehsan; Fardipour, A

doi:10.22055/ivj.2015.8715

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

Document Type : Research Paper

Authors

ehsan zayerzadeh

A Fardipour

10.22055/ivj.2015.8715

Abstract

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.

Keywords

Clostridium perfingens

epsilon toxin

Chromatography

Gel filtration

Subjects