细胞色素P450酶复合组装与代谢行为及其生物分析应用研究

The vivo biological molecules are assembled together in some certain models and play corresponding roles synergistically. The simulation of biological molecular assembly will help us to research life activities and the signal transduction in biological systems, which will improve human health. "Metabolon" mentioned in this project refers to multi-enzyme complexes with metabolic function. "Metabolic channel" is a coupled enzyme reaction with consecutive conversion steps in the metabolon. Metabolon with excellent metabolic activity and stable property will be designed and constructed to research the activity of biological molecules and the properties of electron and energy transfer in the metabolon, revealing the chemical nature and dynamic mechanisms of metabolism. Two-dimensional metabolon will be constructed by graphene and other nano-materials with the help of click reaction and atom transfer radical reaction to study the influences of distribution and interaction of biological molecules on metabolism and explain the metabolic behavior and its dynamic mechanism in the two-dimensional metabolon. On the basis of the above work, the research platform for drug metabolism in vitro will be built to provide new methods and technologies for the research on the formation of metabolon and metabolic channel and metabolic process intervention. The research has great significance in human health, discipline-crossing and the development of bioanalytical chemistry. It will also play an important role in enhancing China's capability of independent innovation in the field of analytical science.

生物体内生物分子具有一定的组装模式并协同发挥作用，模拟生物系统的分子组装将有助于我们探究生命活动和生物系统中的信号传递，继而改善人类健康。本项目提出的"代谢区室"是具有代谢功能的细胞色素P450酶复合组装体，"代谢通道"是指代谢区室内发生的具有连续转换步骤的偶联酶反应过程。本项目设计构建具有优异代谢活性和稳定性质的代谢区室，研究代谢区室内生物分子的组装行为、生物活性、电子转移、能量传递等对其代谢行为的影响，揭示其代谢作用的化学本质和动力学机制；利用石墨烯等纳米材料以及点击化学反应、原子转移自由基反应构建二维代谢区室，阐述二维受限区域内的代谢行为及其动力学机制；在上述研究的基础上，构建药物体外代谢研究平台，为研究代谢区室和代谢通道的形成、干预代谢过程等提供新方法和新技术。本项研究对于人类健康、学科交叉和生命分析化学的发展以及提高我国在分析科学领域中的自主创新能力具有十分重要的意义。

四年来，在基金21375014的资助下，围绕细胞色素P450酶的复合组装、代谢区室构建、代谢区室内酶对药物和环境物质的选择性催化行为及其反应动力学等关键科学问题，开展细胞色素P450酶复合组装及其受限空腔内酶的电荷传输、物质迁移、能量传递、信息传递以及对底物的级联代谢行为的研究，构建了8种基于细胞色素P450酶纳米组装、纳米孔道与硅泡沫限域以及光电驱动的药物代谢系统；研究了双酶在石墨烯纳米笼、纳米金/石墨烯和氧化铝纳米阵列通道内的受限组装及其光电驱动的催化反应动力学，阐述其与生物系统代谢反应的共性；构建了多种生物分子与纳米材料的复合组装方法以及DNA、端粒酶传感新技术，项目成果共发表论文47篇，包括Analytical Chemistry 9篇、Chemical Communications 6篇，Biosensors and Bioelectronics 7篇、Analytica Chimica Acta 5篇、ACS Applied Materials and Interfaces 3篇、Journal of Electroanalytical Chemistry 4篇、Analyst和Microchimica Acta各2篇，Journal of Materials Chemistry A、Langmuir、Talanta、Electrochimica Acta、RSC Advances、Current Analytical Chemistry、Analytical and Bioanalytical Chemistry、IEEE Sensors和《电化学》各1篇。获授权中国发明专利3件（ZL 2014 1 0404833.4；ZL 2012 1 0475240.8和ZL 2014 1 0108594.8）。本项目的成果对于研究与模拟生物系统的代谢与生物转化、药物研发和健康干预等具有十分重要的意义。本项目培养博士生13人（毕业10人），硕士生11人（毕业7名）。在国际、国内学术会议上邀请报告13次，并对美国马里兰大学、加拿大多伦多大学进行了访问。