Comparison of Two Purification Methods for Beta Toxin of Clostridium perfringens Type B

Abstract

Clostridium perfringens is an anaerobic bacterium that among its strains, only types B and C can produce beta-toxin. Beta-toxin plays an important role in many human and animal diseases and has detrimental effects on the intestine. The present study aimed to compare two purification methods for the extraction of beta-toxin from C. perfringens type B. As the first method, beta-toxin was purified using ammonium sulfate precipitation, column chromatography (Sephadex G-25), and ion-exchange chromatography (DEAE Sephadex), while the second method employed affinity chromatography. Hemolysis activity and protein content were measured in each step of purifications. The purity of beta-toxin was monitored in each step using SDS-PAGE. According to the results, a comparison of these two methods indicated that the yield of the first method was 82.8%, and the yield of the second method was 90.1%. The specific activity values for the first and second methods were calculated to be 2368.1 U/mg and 164.5 U/mg, respectively. Our results show that affinity chromatography could be used to purify beta-toxin from clostridium perfringens type B with high purity and specific activity (4370 HU/mg).