Purification, characterization and anticoagulant activity of a proteolyticenzyme from Vespa orientalis venom

The anticoagulant effect of Vespa orientalis venom sac extract (VSE) was at tributed to a proteolytic process, involving mainly coagulation factors VII I and IX [Joshua, H., Ishay, J., 1975. Toxicon 13, 11-20; Korenberg et al., 1988. Toxicon 26, 1169-1176]. Preliminary purification of the proteolytic activity showed the presence of three separate proteases. One of which, pro tease I, was purified. The purified enzyme migrated as a double blind on so dium dodecyl sulfate polyacrylamide gel electrophoresis (SDS PACE). The mol ecular weights of the bands, under reduced conditions were 42 and 44 kD. Bo th bands retained activity after the electrophoretic run. The enzyme hydrol yses bovine factor IX (BFIX), factor X (BFX) and prothrombin. The pH optimu m for the degradation of BFIX was 7.0 and its isoelectric point is above pH 10. The amino acid composition of the protease was determined. (C) 1999 El sevier Science Ltd. All rights reserved.