A scheme of the purification of listeriolysin O produced by L. monocytogenes strain NCTC 7973 was developed. The isolation procedure included the cultivation of the bacteria in heart-brain broth, the concentration of culture liquid free of bacteria with ammonium sulfate, cation exchange chromatography on a column packed with CM-Sepharose and Mono S, gel chromatography on a column packed with Superose 12. The preparation obtained with the use of this procedure was homogeneous, as confirmed by the data of SDS electrophoresis. The protein obtained in this investigation was no different from the protein studied earlier in its physico-chemical properties (molecular weight, heat stability, inhibition with thiole-active compositions, cholesterol, sensitivity to proteolytic enzymes) and corresponded to the characteristics of thiole-dependent hemolysins.
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