Demonstration and isolation of phallolysin, a haemolytic toxin from Amanita phalloides

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Abstract
Phallolysin, a high-molecular weight, thermo- and alcohol-labile toxin with haemolytic activity, was demonstrated in Amanita phalloides. Phallolysin was isolated from cold aqueous toadstool extracts by precipitation with 40% ammonium sulphate, ion-exchange chromatography on DEAE cellulose, gel chromatography on Biogel P-30 and electrofocussing in Ampholine. Electrofocussing in a flat pH gradient resulted in the separation of the gelchromatographically homogeneous phallolysin into phallolysin A (I.P. 8.06) and phallolysin B (I.P. 7.49). Phallolysin A exhibited 70% purity with respect to its behaviour on polyacrylamide gel electrophoresis when stained with Coomassie Brilliant Blue. On the average 0.62 μg, at the best 0.20 μg of protein was equivalent to 1 haemolytic unit (HU). The i.p. LD50 for mice was found to be 1060 (930–1210) HU/kg.