Purification and characterization of Clostridium perfringens beta toxin

A new procedure for the purification of beta toxin from culture supernatant fluid of Clostridium perfringens was established. The procedure consists of ammonium sulfate fractionation, affinity chromatography on zinc-chelate Sepharose and gel filtration on Toyopearl HW 60. Beta toxin was purified about 460-fold from the ammonium sulfate fraction with a yield of about 60% in terms of lethality of the toxin. The molecular weight of the toxin, determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and sucrose gradient centrifugation, was approximately 40,000. The isoelectric point was 5.6. The minimal necrotic dose for guinea pigs was approximately 2 ng. The 50% lethal doses for adult mice were 310 ng/kg and 4.5 micrograms/kg, when injected i.v. and i.p., respectively.