【摘要】：白屈菜紅堿(CHE)是一種生物活性豐富的生物堿,可抗癌、殺菌,并具有良好的滅螺特性,但在植物體內(nèi)含量較少、分離困難,因此它的高效分離一直備受人們的關(guān)注。本文合成了一種新型的核-殼-殼結(jié)構(gòu)的分子印跡磁性聚合物微球,研究了該微球?qū)Π浊思t堿的吸附動(dòng)力學(xué)與熱力學(xué),并進(jìn)行了白屈菜紅堿的高效分離研究。本文還深入探討了白屈菜紅堿與螺類功能蛋白——鑰孔戚血藍(lán)蛋白(KLH)的相互作用機(jī)理,為白屈菜紅堿作為天然滅螺藥物的開發(fā)提供了有益的參考。具體研究?jī)?nèi)容如下:(1)以白屈菜紅堿為模板,四乙烯基吡啶為功能單體,Fe3O4為磁性基質(zhì),采用自由基聚合法合成了白屈菜紅堿分子印跡磁性聚合物微球(CHE-MMIPs),并對(duì)其進(jìn)行了形貌表征。結(jié)果表明,該微球具有較窄的粒徑分布(約240nm),良好的磁響應(yīng)度(40emu/g),較好的單分散性。(2)考察了CHE-MMIPs的吸附效率。結(jié)果表明,CHE-MMIPs最大的吸附容量為12mg/g(6 h),吸附過程符合Langmuir等溫吸附模型及準(zhǔn)二級(jí)動(dòng)力學(xué)方程,屬單層吸附。(3)利用上述實(shí)驗(yàn)純化的CHE,采用熒光光譜與分子對(duì)接技術(shù)研究了CHE對(duì)KLH的熒光猝滅機(jī)制。結(jié)果表明,CHE可以顯著猝滅KLH的熒光,其猝滅機(jī)制為靜態(tài)猝滅。在293K、298K、303K時(shí),CHE與KLH的結(jié)合常數(shù)分別為4.72×105、5.06×105、1.01×106L/mol。體系的熱力學(xué)參數(shù)ΔG298K、ΔH298K、ΔS298K的值分別為 32.54 kJ/mol、 58.78 kJ/mol、和 88.05 J/mol·K。在結(jié)合過程中主要的作用力為范德華力和氫鍵。采用分子對(duì)接技術(shù)獲得了CHE與KLH結(jié)合的具體位置、位點(diǎn)數(shù)及結(jié)合親和力的大小。發(fā)現(xiàn)KLH中的Cu位點(diǎn)并不是CHE與KLH的潛在結(jié)合位點(diǎn)。
[Abstract]:(CHE) is a kind of alkaloid with rich biological activity, it can be anticancer, bactericidal and has good snail killing characteristics. However, it has little content in plants and is difficult to be separated. Therefore, its high efficiency separation has been paid close attention by people all the time. In this paper, a novel core-shell structure molecularly imprinted magnetic polymer microspheres was synthesized. The adsorption kinetics and thermodynamics of the microspheres for chordythrine were studied, and the high efficiency separation of alchlor alkaloids was studied. The mechanism of interaction between chordythrine and keyhole hemocyanin (KLH), a functional protein of snail, was also discussed in this paper, which provided a useful reference for the development of chordythrine as a natural snail killing drug. The main contents are as follows: (1) the molecular imprinted magnetic polymer microspheres (CHE-MMIPs) were synthesized by free radical polymerization using tartarine as template, tetravinylpyridine as functional monomer and Fe3O4 as magnetic matrix. Its morphology was characterized. The results showed that the microspheres had narrow particle size distribution (about 240nm), good magnetic responsivity (40emu/g) and good monodispersity. (2) the adsorption efficiency of CHE-MMIPs was investigated. The results show that the maximum adsorption capacity of CHE-MMIPs is 12mg/g (6-h), adsorption process accords with Langmuir isotherm adsorption model and quasi-second order kinetic equation, and belongs to monolayer adsorption. (3) the CHE, purified from the above experiments belongs to monolayer adsorption. The fluorescence quenching mechanism of CHE on KLH was studied by fluorescence spectroscopy and molecular docking technique. The results showed that CHE could significantly quench the fluorescence of KLH, and its quenching mechanism was static quenching. At 293K, 298K and 303K, the binding constants of CHE and KLH were 4.72 脳 105,5.06 脳 105,1.01 脳 106L mol. The thermodynamic parameters 螖 G298K, 螖 H298K and 螖 S298K are 32.54 kJ/mol, / mol 58.78 kJ/mol, and 88.05 J / mol 路K. The main forces in the bonding process are van der Waals force and hydrogen bond. The binding position, the number of sites and the binding affinity of CHE and KLH were obtained by molecular docking technique. It was found that the Cu site in KLH was not a potential binding site between CHE and KLH.
【學(xué)位授予單位】：湖南理工學(xué)院
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：O631.3

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