基于热诱导的鱼鳞胶原去折叠及热聚集调控机制研究

In the recent years, studies have proved that collagen has remarkable potential for use in many fields. Heat treatment is the main method used to produce collagen. As an economic and scientific problem, collagen unfolding and aggregation behavior resulting from heat treatment are key factors affecting the product’s quality. Based on the "turning waste into avail" concept, marine bio-processing by-product protein resources were used for this study, and chromatography, spectroscopy, mass spectrometry and electron microscopy technology were used to study the regulatory mechanisms of scale collagen subjected to heat-induced treatment. The structure and function of scale collagen have been investigated during the heat-inducted unfoldment. The variations in the structure and function from scale collagen in different states of aggregation and de-aggregation will also be investigated. Then, a kinetic model will be established for collagen unfolding to analyze the manner in which unfolding takes place and the variation with respect to hydrogen, hydrophobic bonds, and other factors. The kinetic model for collagen de-aggregation will also be established to analyze the regulatory mechanism of natural product inhibitors. In summary, the project will assess the regulatory mechanisms affecting the unfolding and aggregation of scale collagen subjected to heat-induced treatment from the perspectives of molecular biology, biochemistry, biophysics, stoichiometry and others. These results will provide important theoretical guidance for and a technical approach to the research and development of collagen. It is of great significance in the perfection of marine biology resource theory and in the development and utilization of marine drugs, food, biomedicine and cosmetics.

近年来，胶原蛋白在多个领域中展示出巨大的应用前景。热处理是胶原蛋白主要的生产技术手段，热处理诱导的胶原去折叠行为和胶原热聚集行为是影响胶原蛋白质量的关键因素，是一个重要的经济和科学问题。本项目基于“变废为宝”的理念，选择海洋生物加工副产物资源为研究对象，以热诱导鱼鳞胶原调控机制为研究目标，采用色谱、光谱、质谱、电镜等组合技术手段，深度研究鱼鳞胶原热诱导去折叠的结构与功能变化特点，鱼鳞胶原热聚集与解聚集的结构与功能变化特点。构建胶原去折叠动力学模型，分析胶原去折叠途径、氢键、疏水区的变化规律。构建胶原抑制热聚集的动力学模型，分析小分子抑制剂的调控作用，从分子生物学、生物化学、生物物理学和化学计量学等多层面、多角度探讨热诱导鱼鳞胶原去折叠、热聚集的调控机制，为胶原蛋白的研究和开发提供理论指导和技术支撑，对海洋生物资源学理论的完善和海洋药物、食品、化妆品、生物医学的有效开发与利用具有重要的意义。

热处理诱导的胶原热聚集行为是影响胶原蛋白质量的关键因素，是一个重要的经济和科学问题。本项目选择鱼鳞胶原为研究对象，研究热诱导鱼鳞胶原调控机制。以鱼鳞为原料，制备鱼鳞胶原和鱼鳞胶原亚基α1和α2，并完成样品的结构鉴定、功能性、流变性和热稳定性的表征。实验结果表明，鱼鳞胶原是一种良好的食品、化妆品和生物功能材料原料。采用光谱协同电镜组合技术，阐释鱼鳞胶原热聚集与解聚集过程中，结构与流变特性的变化规律。在此基础上，利用荧光探针法、浊度法及Boltzmann函数拟合法，构建鱼鳞胶原热聚集/解聚集的动力学模型，分析小分子化合物调控胶原的热聚集行为。实验结果表明，没食子酸和白藜芦醇能够调控鱼鳞胶原热聚集。没食子酸在鱼鳞胶原热聚集的“起始阶段”，抑制热聚集的效果优于白藜芦醇；而白藜芦醇在胶原热聚集的“快速生长阶段”；抑制热聚集的效果优于没食子酸。建立热诱导鱼鳞胶原去折叠计算机动力学模型，模拟热诱导胶原去折叠过程，二级结构、氢键的变化规律。计算机模拟结果表明，当温度高于热变性温度时，随着温度的升高，类胶原二级结构由无规则coil转变为bend，类胶原A-B链间、B-C链间稳定氢键数目减少，类胶原分子间的氢键作用减弱。当温度高于100℃时，部分类胶原的二级结构转变成anti-parallel beta-sheet，类胶原分子开始解离。故，本项目研究对海洋胶原蛋白精深加工理论的完善具有重要的意义。