Simplified method for purification of Clostridium perfringens type A enterotoxin.
Purification of Clostridium perfringens type A enterotoxin from sporulated cells was simplified. The method consisted of precipitation of the enterotoxin from the extract of sonically treated cells at 40% saturation of ammonium sulfate at pH 7, differential solubilization in 0.02 M phosphate buffer, pH 6.7, and repeated gel filtration on Sephadex G-200. The purified enterotoxin was at least 98% pure in ultracentrifugation, polyacrylamide gel electrophoresis, and agar gel double diffusion. Recovery was over 74% from the sporulated cell extract. The toxin had biological activities of at least 4,700 mouse intravenous minimal lethal doses/mg of N, 3,900 capillary permeability-increasing U/mg of N in the guinea pig skin, and 210 rabbit intestinal loop distension U/mg of N. The toxin, containing no hexose, lipid, or nucleic acid, appeared to be identical in sedimentation constant, isoelectric point, and ultraviolet absorption spectrum to the toxin purified previously by different procedures.
D000645:Ammonium Sulfate; D000818:Animals; D001426:Bacterial Proteins / Q000032:analysis; D002199:Capillary Permeability / Q000187:drug effects; D002850:Chromatography, Gel; D003016:Clostridium perfringens; D003470:Culture Media; D004591:Electrophoresis, Polyacrylamide Gel; D004768:Enterotoxins / Q000302:isolation & purification* / Q000494:pharmacology / Q000633:toxicity; D006168:Guinea Pigs; D005779:Immunodiffusion; D007422:Intestines / Q000187:drug effects; D007525:Isoelectric Focusing; D008055:Lipids / Q000032:analysis; D008722:Methods; D051379:Mice; D009500:Neutralization Tests; D011817:Rabbits; D012882:Skin Tests; D013056:Spectrophotometry, Ultraviolet; D013171:Spores, Bacterial; D014461:Ultracentrifugation
D001426:Bacterial Proteins; D003470:Culture Media; D004768:Enterotoxins; D008055:Lipids; D000645:Ammonium Sulfate