Production and purification of beta toxin from Clostridium perfringens type C by chromatography
2012
Ehsan Zayerzadeh
Clostridium perfringens is an anaerobic, spore-forming, gram positive pathogen of humans and domestic animals. The virulence of C. perfringens is largely attributable to its prolific secretion of toxins. Beta toxin of Clostridium perfringens type C is necrotizing and lethal. The toxin has been thought to be a main factor in necrotic enteritis caused by C.perfrirtgens type C. In the present study beta-toxin was purified about 77.8-fold from culture supematant fluid of Clostridium perfringens type C with a yield of about 16.7% in terms of biologically active beta-toxin. The purification procedure involved ammonium sulfate precipitation, Cation exchange chromatography (CM-Sepharose), anion exchange chromatography (DEAE-Cellulose) and gel filtration through Sephadex G- 100. Minimal lethal dose of primary culture supernatant was determined approximately 1/2000 for adult mice. Best percentage of ammonium sulphate for precipitation was determined about 50%. The molecular weight of the toxin, determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis was approximately 37 KD. The 50% lethal doses for adult mice were 2.21 Og/kg. Biological activity of beta toxin was decreased following incubation period in 60 Ί C.
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