Korean Journal of Veterinary Research 1999;39(5):1028-1032.
Production and partial purification of Staphylococcus aureus alpha toxin
Hee-myung Park, Tae-ho Oh, Hong-ryul Han
Department of Veterinary Internal Medicine, College of Veterianry Medicine, Seoul National University
Abstract
Alpha toxin of S aureus has cytolytic activity respectively. This antigen has been received the most attention since it is a major virulence factor in pathogenesis of staphylococcal mastitis. Thus, alpha toxin has been focused as potential candidate of vaccine to minimize mastitis in cows. The purpose of this study was to develop a simple, efficient production and purification methods of sufficient amount of alpha toxin antigen from S aureus. Alpha toxin production measured by hemolytic activity was the highest at 18 hrs postinoculation in yeast extract culture medium supplemented with thiamine, nicotinic acid and casamino acid. Alpha toxin was purified by ammonium sulfate precipitation (65%) and ultrafiltration. Molecular weight of the toxin was 33 kDa in the analysis with SDS-PAGE. Conclusionally, when alpha toxin was included in the vaccine, the optimal harvest time of alpha toxin was at 18 hrs after inoculation in yeast extract medium supplemented with thiamine and nicotinic acid.
Key Words: alpha toxin, Staphylococcus aureus, production, purification


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