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.
D000135:Acid Phosphatase / Q000378:metabolism; D000596:Amino Acids / Q000032:analysis; D000645:Ammonium Sulfate; D000818:Animals; D000833:Animals, Suckling; D001427:Bacterial Toxins / Q000032:analysis / Q000276:immunology / Q000302:isolation & purification* / Q000633:toxicity; D002135:Calcium-Binding Proteins; D002614:Chelating Agents; D055598:Chemical Phenomena; D002627:Chemistry, Physical; D002846:Chromatography, Affinity; D004591:Electrophoresis, Polyacrylamide Gel; D006168:Guinea Pigs; D006636:Histamine Release / Q000187:drug effects; D007525:Isoelectric Focusing; D051379:Mice; D008970:Molecular Weight; D010738:Type C Phospholipases
D000596:Amino Acids; D001427:Bacterial Toxins; C031884:CPB protein, Clostridium perfringens; D002135:Calcium-Binding Proteins; D002614:Chelating Agents; D000135:Acid Phosphatase; D010738:Type C Phospholipases; C040649:alpha toxin, Clostridium perfringens; D000645:Ammonium Sulfate