Scorpion parts of the separation and analysis of anticoagulant activity and kinetics of enzymatic
|School||Beijing University of Traditional Chinese Medicine|
|Course||Chinese medicine pharmacy|
|Keywords||Dynamics Anticoagulant Enzymatic Scorpion Thermodynamics|
Objective To study the Scorpion enzymatic hydrolysis , purification anticoagulant activity sites , explore the enzymatic reaction mechanism, Calculating kinetic and thermodynamic parameters . Methods APTT, fibrinogen tablet dissolved circle area as an index that combines optical density method and protein hydrolysis is preferably pepsin, trypsin digestion process conditions Scorpion determine the reaction temperature , time and enzyme-substrate ratio and other parameters . Using gel filtration chromatography purified pepsin hybrid polypeptide and its preliminary quality standard . Based on Michaelis-Menten equation , study Scorpion enzymatic kinetics , calculated on the basis of the rate constants , activation energy and other kinetic , thermodynamic function value. 1 ) Scorpion optimal enzymatic process: Scorpion powder 37 ℃ artificial gastric juice added 3.0% ( enzyme substrate ratio ) pepsin 3h, the residue into 37 ℃ artificial intestinal Following 5% ( enzyme substrate ratio ) trypsin digestion 4h. 2 ) pepsin was eluted with deionized water , followed by Sephadex Sephadex G-100, SephadexG-50 or SephadexG-25 chromatography , eluent collected anticoagulant efficacy of the active site screened QF50 -3 - ③. 3 ) Draw Scorpion pepsin hydrolysis system curve under different conditions , the kinetic equation is derived (dH) / (dt) = αS 0e-bH = (19.525E0 1.2666S0) e-0.098h, and to obtain the system corresponding thermodynamic and kinetic constants Km = 0.00363 (g · L-1), Ea = 34.6577 (KJ · mol-1). Conclusions This paper establishes the Scorpion extraction process route for animal drugs enzymatic extraction provides a research model ; Purification Scorpion anticoagulant active site for the Scorpion anticoagulant efficacy studies laid the material foundation ; conduct enzymatic kinetics and thermodynamic studies , so as Scorpion enzymatic process design and conditions of the investigation provides the basis for theoretical data .