Purification and Some Properties of a Bacillus cereus Mouse Lethal Toxin

Abstract

A mouse lethal toxin (MLT) produced by Bacillus cereus isolated in vomiting-type food poisoning was purified by chromatography on DEAE-Sephadex A-25 followed by gel filtration on Sephadex G-75. Purified MLT possessed a molecular weight of 33, 000-34, 000. It showed mouse lethality and hemolytic (HL) activity on sheep and rabbit erythrocytes; the latter erythrocytes were more weakly hemolyzed than the former ones. However, fluid accumulation in mouse ligated intestinal loops was not induced by purified MLT at the highest concentration used. Both MLT and HL activities were stable at pH 6-9, during storage at -20°C for 8 weeks, and resistant to papain, cholesterol, lecithin, and dithiothreitol treatments. Most activity was lost during storage at 4°C or 25°C for 2 weeks or upon treatment with trypsin, trypanblue, or ethanol. The activities were resistant to heating at 37°C for 5 min, less resistant at 98°C for 5 min, and sensitive at 60°C for 5 min. It can be concluded from the results that MLT is different from the diarrheagenic toxin produced by B. cereus isolated in diarrheal-type food poisoning, but is similar to, if not identical, hemolysin II.