Simplified Purification and Biophysicochemical Characteristics of Kanagawa Phenomenon-Associated Hemolysin of Vibrio parahaemolyticus

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Abstract
Kanagawa phenomenon-associated hemolysin (K-hemolysin) was purified by Sephadex gel and ion-exchange column chromatography after the culture supernatant was adsorbed on and eluted from DEAE-Sepharose CL-6B and acid precipitated. K-hemolysin was a heat-stable and trypsin-susceptible protein with an apparent MW of 44,000, the subunit of which was 22,000. The isoelectric point was 4.9. The minimum hemolytic dose was 0.1 .mu.g/ml. The LD50 by i.v. injection was 1.4 .mu.g. EM of the small intestine of suckling mice orally challenged with the highest dose (50 .mu.g) showed disappearance of epithelial cell microvilli and structural disturbances of the endoplasmic reticulum and mitochondrial swelling. One blueing dose representing permeability factor activity was 0.3 .mu.g and positive reaction in the rabbit ileal loop appeared at above 125 .mu.g. An antibody appeared in patients which neutralizes K-hemolysin during the course of the disease. K-hemolysin may play a most significant role in the pathogenesis of this enteric human disease.