C60富勒烯模拟酶化学发光评价体系的构建及其分析应用研究

In recent years,colorimetric method using nanoparticle-based mimetic enzymes has been widely used in biochemical, environmental and clinical analysis. However, chemiluminescent method using nanoparticle-based mimetic enzyme has seldom been reported.Chemiluminescence is sensitive, simple and cheap, etc.In this project, we will evaluate the mimetic catalytic properties of C60 fullerenes by chemiluminescent method . Their catalytic properties will be investigated by considering the dependence of catalytic behavior of fullerenes on pH, temperature and substrate concentration and the chemiluminescence evaluation system will be constructed taking into account the values of maximum reaction velocity, Michaelis constant and catalytic constant. Electrochemical methods, UV/Vis absorption spectroscopy, X-ray photoelectron spectroscopy, chemiluminescence spectroscopy and electron spin resonance spin-trapping will be used to explore the catalytic mechanisms. Highly sensitive analytical methods for substrates will be established based on the chemical properties of the substrate and the catalytic mechanism of fullerene. The smooth implementation of this project will increase understanding of the biocatalytic properties of nanomaterials and reveal the advantages of chemiluminescence-based evaluation of nanoparticle-based mimetic enzymes.This project will expand the theoretical research of the nanoparticle-catalyzed chemiluminescence, which is of scientific importance and it will provide a basis for the development of new chemical sensor technology and biochemical environment analysis technology.

近几年来，基于纳米粒子模拟酶所构建的比色分析法已在生化、环境及临床分析中得到广泛应用。然而，基于纳米粒子模拟酶构建化学发光分析法鲜见报道。化学发光具有灵敏、简便及廉价等优点。本项目拟利用化学发光法评价C60富勒烯模拟酶催化特性，从C60富勒烯的催化行为对温度、环境pH和底物浓度等条件的依赖性考察其催化性能，基于其最大反应速率(Vmax)、米氏常数(Km)和催化常数（Kcat）等参数构建化学发光评价体系。借助电化学方法、紫外-可见光谱、化学发光光谱、X-射线光电子能谱和顺磁共振技术等手段揭示其催化机理。基于底物的化学性质和C60富勒烯的催化机理构建其高灵敏的分析方法。项目的顺利实施将进一步加深人们对纳米材料的生物催化特性的认识并了解基于化学发光法评价纳米粒子模拟酶的优势，对深化纳米材料催化化学发光的基础理论研究具有重要的科学意义，为发展新型化学传感技术和生化环境分析技术提供一定的理论基础。

基于纳米粒子模拟酶构建的比色分析法是近几年来科技工作者关注的焦点问题之一。化学发光具有灵敏、简便及廉价等优点。然而，基于纳米粒子模拟酶构建化学发光分析法鲜见报道。本项目利用化学发光法评价C60富勒烯模拟酶催化特性，主要研究内容有：（1）C60富勒烯的改性和处理；（2）C60富勒烯的表征；（3）C60富勒烯的催化性能研究和新化学发光体系的构建；（4）C60富勒烯做为模拟酶评价体系的构建；（5）利用C60富勒烯构建新分析方法的生物和环境样品测定和（6）C60富勒烯催化化学发光机理的研究。实验结果表明，利用甘氨酸修饰C60富勒烯可极大的改善其水溶性。将H2O2-鲁米诺发光体系为模型构建C60富勒烯的过氧化物模拟酶评价体系，将该发光体系与葡萄糖氧化酶(GOx)催化溶解氧氧化葡萄糖产生过氧化氢反应相结合，可构建一种一个简单、灵敏葡萄糖化学发光传感体系。该项目的顺利实施充分展现了基于化学发光法评价纳米粒子模拟酶的优势，对深化纳米材料催化化学发光的基础理论研究具有重要的科学意义，为发展新型化学传感技术和生化环境分析技术提供一定的理论基础。