Isolation and Purification of Thrombin-like Enzyme from the Venom of Agkistrodon halys Pallas / 中国药房

ABSTRACT

OBJECTIVE:To study a simple method for the isolation and purification of thrombin-like enzyme (TLE) from the venom of Agkistrodon halys Pallas. METHODS:DEAE-Sephadex A-25 vs. Sephadex G-25 chromatography in the isolation and purification of TLE from the venom of Agkistrodon halys Pallas was analyzed. RESULTS:TLE was isolated from the venom of Agkistrodon halys Pallas. SDS-PAGE electrophoresis appeared as a strap with its molecular weight at about 35.5 kDa, meeting electrophoresis purity standard. Physical-chemical character assay showed that the TLE has hemostasia activity in vitro with its specific activity at about 12.57IU?mg-1, while the arginine esterase activity was about 137.65IU?mg-1 measured by BAEE method. PMSF and EDTA were used in the inhibition experiment on this enzyme, and the enzyme was proved to be serine protease other than metalloproteinase. CONCLUSION:The methods can be used for the isolation and purification of thrombin-like enzyme (TLE) from the venom of Agkistrodon halys Pallas.