新型多孔金属氧化物对胰蛋白酶的高效亲和固定化与导向性分离研究

项目名称： 新型多孔金属氧化物对胰蛋白酶的高效亲和固定化与导向性分离研究

项目编号： No.21301001

项目类型： 青年科学基金项目

立项/批准年度： 2014

项目学科： 数理科学和化学

项目作者： 李士阔

作者单位： 安徽大学

项目金额： 25万元

中文摘要： 本项目针对当前酶固定化方法中存在的选择性差，固定化酶易失活等关键难题，利用设计合成的具有核壳或蛋黄-蛋壳结构的Fe3O4@NiO、Fe3O4@CuO超顺磁性新型多孔金属氧化物为载体。以NiO、CuO等多孔金属氧化物表面较为丰富的Ni2+,Cu2+等金属离子与具有His标签的胰蛋白酶的高效亲和作用，构建选择性高，活性、稳定性增强高效亲和胰蛋白酶固定化新方法。结合超顺磁性纳米载体的特点，发展具有导向分离特征的胰蛋白酶快速分离纯化技术。阐述新型多孔金属氧化物载体结构、成分等微观形态与胰蛋白酶的固载含量、活性、稳定性及循环使用效率间的关系。阐明新型多孔金属氧化物对胰蛋白酶稳定性、活性增强的机制，为设计、合成性能更加优越，用途更为广泛的蛋白等生物分子的快速分离纯化技术及细胞活体固定化方法提供启示。

中文关键词： 比表面积；纳米结构；生物分子吸附；分离纯化；

英文摘要： This research aims at solving the current special challenges in enzyme immobilization, such as poor selectivity, activity loss of the immobilized enzyme. This novel immobilization method for trypsinase with high immobilized capacity, biological activity, and stability is proposed based on the superparamagnetic Fe3O4@CuO, Fe3O4@NiO core-shell or yolk-shell porous metal oxides, which will be used as high efficient matrixes for trypsinase immobilization by the affinity interactions between Cu2+, Ni2+ ions on the porous shell surface and the His-Tagged residues of trypsinase. By means of the superparamagnetic properties of the novel matrixes, the trypsinase will be rapidly oriented collected from the reaction solutions. This novel enzyeme immobilized protocol will powerfully promote the developments for high efficient recyclable catalysis. The researches will show the relationship between the microstructure, composition of the novel porous metal oxide matrixes and the immobilization capacity, biological activity, stability and recyclable catalysis efficiency of trypsinase, as well as explore the enhancement mechanism of biological activity, stability of the immobilized trypsinase. These results may find potential applications in rapid separation and purification of proteins and cells immobilization.

英文关键词： specific surface area；nanostructure；immobilization of biomolecules；separation and purification；