Isolation and some properties of an enterotoxin produced by Bacillus cereus

Extracellular proteins produced by Bacillus cereus B-4ac were separated by chromatography on Amberlite CG-400, QAE-Sephadex, Sephadex G-75, and hydroxylapatite. A fraction, containing three detectable antigens, obtained from chromatography on hydroxylapatite caused fluid accumulation in ligated rabbit ileal loops, was dermonecrotic to rabbit skin, was cytotoxic to cultured cells, and was lethal to mice after intravenous injection. Two other fractions obtained from chromatography on hydroxylapatite showed essentially no toxic activity when tested individually. Each nontoxic fraction contained two of the three proteins present in the toxic material. When the two nontoxic fractions were combined, activity in all of the biological assays was observed. Antiserum against either of the nontoxic fractions neutralized the dermonecrotic response of the combined material. These results suggest that all of these biological activities probably are due to a single entity and that more than one component probably comprise the toxic entity.