Dissertation
Dissertation > Medicine, health > Chinese Medicine > Of Pharmacy > Traditional Chinese medicine chemical

Scorpion parts of the separation and analysis of anticoagulant activity and kinetics of enzymatic

Author XuWenBo
Tutor WangYuRong
School Beijing University of Traditional Chinese Medicine
Course Chinese medicine pharmacy
Keywords Dynamics Anticoagulant Enzymatic Scorpion Thermodynamics
CLC R284
Type Master's thesis
Year 2010
Downloads 107
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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 .

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