Separation and Purification of Haemocoagulase from Agkistrodon Blomhoffii in Changbai Mountain
- VernacularTitle:长白山白眉蝮蛇血凝酶的分离纯化研究
- Author:
Shoushi JIA
;
Mengmeng WANG
- Keywords:
Agkistrodon blomhoffii in Changbai Mountain;
Haemocoagulase;
Separation and purification
- From:
China Pharmacist
2017;20(2):212-215
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To establish a method for the separation and purification of haemocoagulase from agkistrodon blomhoffii in Changbai Mountain. Methods: An enzyme component with clotting activity was isolated from the venin of agkistrodon blomhoffii in Changbai Mountain by gel filtration, anion-exchange chromatography and heparin-Sepharose affinity chromatography. SDS-Page and RP-HPLC were used to determine its purity, SDS-Page was applied to study its action mode with bovine fibrinogen, HPSEC was used to determine the molecular weight, an IEF method was employed to detect its isoelectric point, and Lowry method was used for the deter-mination of protein concentration. Results:One haemocoagulase was purified from agkistrodon in Changbai Mountain. SDS-Page dis-played one band, and HPLC showed one single chromatographic peak. The haemocoagulase acted only on α chain of fibrinogen. Its molecular weight was 32. 2kD with isoelectric point of 5. 21. The enzyme had clotting activity in vitro. Conclusion:The method can be used for the separation and purification of haemocoagulase from agkistrodon blomhoffii in Changbai Mountain.
