Identification and Characterization of a Thrombin-like enzyme isolated from Deinagkistrodon acutus venom

摘要

A thrombin-like enzyme was isolated from Deinagkistrodon acutus venom by a combination of affinity and ion exchange chromatography. The enzyme presented ~40 kD by reducing SDS-PAGE analysis, ~34 kD by MALDI-TOF-TOF-MS analysis, and was shown to be glycosylated with ~6 kD O-linked carbohydrates, with pI ~4.8±0.2. The enzyme had optimal esterase activity on BAEE and the most serious degradation at pH 9, displayed maximum catalytic rate at ~50. Its hydrolytic activity was °C inhabited by Aprotinin, PMSF, AEBSF and Benzamidine. The enzyme also showed fibrinogen (Fg) clotting activity and amidase activity on DL-BAPA. N-terminal 15 amino acid sequence, PMF analysis and amino acid combination analysis revealed that it has high similarity with other thrombin like enzymes from snake venoms.