基于蛋白质组学和时空动态的微波促进蛋白质糖基化反应的机制研究

前期研究发现，微波会促进蛋白质的糖基化反应，且反应位点、反应程度与蛋白质结构及微波加热不均匀性密切相关，但是其促进机制尚不清楚。针对这一科学问题，本项目拟从蛋白质组学和时空动态变化角度出发，以卵清蛋白和葡萄糖为对象，建立干样和湿样的圆柱模型，采用荧光光纤温度传感器的无接触小型探针和红外热像仪监测其温度分布；通过样品分割、电泳、色度、DSC、光谱技术等分别从反应程度和理化性质方面监测反应历程；采用胶上酶解、LC FTICR MS、ETD MSMS、18氧稳定同位素标记等方法对蛋白质糖基化产物的反应位点进行精确定位和相对定量分析；揭示微波场内蛋白质糖基化反应的规律，建立不同时空条件下微波条件-温度分布-糖基化反应之间的关系，从分子水平和时空动态变化方面阐述微波促进蛋白质糖基化反应的内在机制。本项目成果为实现微波场中蛋白质糖基化反应的定向调控奠定理论基础，对食品科学的进步与发展具有重要意义。

Previous research indicated that microwave could promote the glycation reaction of the protein，and the reaction degree of each site is highly related with protein structure and the non-uniform temperature distribution of microwave heating. However, the promotion mechanism is yet unknown. According to the scientific problem, from the perspective of proteomics and space-time dynamics change during microwave processing, this project aims to investigate the promoted mechanism of microwave on the glycation reaction between ovalbumin and glucose under different time and space by establishing cylindrical dry and wet model. The temperature distribution of microwave heating sample was determined by fluorescence optical fiber temperature sensor with no contact mini probe and infrared thermographic imager. After transverse and longitudinal split of sample, the glycation reaction process under different space and time is supervised by studying the reaction degree and physico-chemical properties of products through electrophoresis, chromaticity, differential scanning calorimetry (DSC), and spectrum analysis. The reaction site, precise identification and relative quantification of glycation products are analyzed by means of proteomics, such as digestion in gel, LC FTICR MS, ETD-MSMS, 18O stable isotope labeling, et al. The rule of glycation reaction is revealed in microwave field. The relationship among microwave conditions, temperature distribution, and glycation reaction is established under different space and time. Then the mechanism of microwave promoting glycation reaction of protein based on proteomics and space-time dynamics. This research can provide theoretical basis for the directional control of protein glycation reaction in microwave field. And it may play a profound role in the development and progressing in food science.