Purification and physicochemical characterization of a serine protease with fibrinolytic activity from latex of a medicinal herb Euphorbia hirta

被引：58
作者：
Patel, Girijesh Kumar [1 ]
Kawale, Ashish Ashok [1 ]
Sharma, Ashwani Kumar [1 ]
机构：
[1] Indian Inst Technol Roorkee, Dept Biotechnol, Roorkee 247667, Uttar Pradesh, India
来源：
PLANT PHYSIOLOGY AND BIOCHEMISTRY | 2012年 / 52卷
关键词：
CD studies; Euphorbia hirta; Euphorbiaceae; Fibrinogenolytic activity; Fibrinolytic activity; Kinetic studies; WESTERN DIAMONDBACK RATTLESNAKE; CORDYCEPS-MILITARIS; CROTALUS-ATROX; SNAKE-VENOM; ENZYME; QUANTITIES; MUSHROOM; MILII;
D O I：
10.1016/j.plaphy.2011.12.004
中图分类号：
Q94 [植物学];
学科分类号：
071001 ;
摘要：
A 34 kDa serine protease, designated as hirtin, with fibrinolytic activity was purified to homogeneity from the latex of Euphorbia hirta by the combination of ion exchange and gel filtration chromatography. The N-terminal sequence of hirtin was found to be YAVYIGLILETAA/NNE. Hirtin exhibited esterase and amidase activities along with azocaseinolytic, gelatinolytic, fibrinogenolytic and fibrinolytic activities. It preferentially hydrolyzed A alpha and alpha-chains, followed by B beta and beta, and gamma and gamma-gamma chains of fibrinogen and fibrin clot respectively. The optimum pH and temperature for enzyme activity was found to be pH 7.2 and 50 degrees C respectively. Enzymatic activity of hirtin was significantly inhibited by PMSF and AEBSF. It showed higher specificity for synthetic substrate p-tos-GPRNA for thrombin. The CD spectra of hirtin showed a high content of beta-sheets as compared to alpha-helix. The results indicate that hirtin is a thrombin-like serine protease and may have potential industrial and therapeutic applications. (C) 2011 Elsevier Masson SAS. All rights reserved.
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收藏
页码：104 / 111
页数：8
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