Purification And Characterization Of A Fibrinolytic Enzyme From Agkistrodon Halys Pallas Venom

Objective:To investigate the purification method of a fibrinolytic enzyme(FLE) from the venom of Agkistrodon blomhoffi brevicaudus and research some biochemistral characterizations.Method: A fibrinolytic enzyme(FLE) was purified from the venom of Agkistrodon blomhoffi brevicaudus by ion-exhange chromatography on a DEAE Sepharose CL-6B column ,followed by gel filtration chromatography on HiPrep Sephacryl S 100. The molecular weight was estimated with SDS-PAGE. The pI was determined through isoelectric focusing. Amino acid composition assay was carried out on a Hitachi 835-50 mould automatic amino acid analyzer .Result : The purified FLE was shown to be homogeneous in SDS—PAGE, which acted as an anticoagulant in vitro. Its molecular weight was estimated to be 59,100 D and isoelectric point was 4.98 . According to the result of amino acid composition assay, it has a number of acidic amino acid, which is consistent with the pI we detected . Through the research of pharmacology, we found it possessed low haemorrhagic activity . FLE belongs to metallic protein and can inhabit the growth of cancer in high concentration.Conclusion : We separate a fibrinolytic enzyme(FLE) which is a acid protein and has low haemorrhagic activity from the venom of Agkistrodon blomhoffi brevicaudus.