MOFs原位自封装固定Burkholderia pyrrocinia源脂肪酶的分子互作机制

采用MOFs材料原位自封装制备固定化脂肪酶是增强酶分子结构刚性从而改善其稳定性，提高固定化酶活性的有效途径，而MOFs材料影响脂肪酶分子构象及固定化过程中的分子互作机制并不清楚。鉴于脂肪酶的“盖子”结构域对其催化活性的显著影响，本项目拟以具有典型“盖子”结构的Burkholderia pyrrocinia源脂肪酶为研究对象，从脂肪酶“盖子”结构运动自由度的角度，通过结构解析结合分子对接和量子计算，探究“盖子”结构对酶分子催化特性的影响；从MOFs-酶分子结合紧密性的角度，明确MOFs影响“盖子”结构运动自由度的变化规律，揭示lipase@MOFs活性变化的内在机制；从固定化酶分子所处的微环境的变化角度，探寻“盖子”结构存在形式差异引起lipase@MOFs分子所处微环境的变化规律，明确MOFs原位自封装脂肪酶的分子互作机制。项目为固定化酶催化机制研究及酶学提供理论依据和科学参考。

The in-situ self-encapsulation of lipase in metal-organic frameworks (MOFs) for immobilization is an effective way to enhance the rigidity of the enzymatic structure, thus improving its stability and catalytic performance of the immobilized enzyme. However, how the MOFs affect the conformation of the enzyme and the molecular interaction mechanism during immobilized process is still unclear. In view of the "lid" structure domain of lipase plays a vital role on its catalytic activity, the present project plan to take lipase from Burkholderia pyrrocinia which has a typical "lid" structure as the model, from the perspective of degrees of freedom of "lid" structure motion, through lipase structure analysis combined with molecular docking and quantum computing methods, explore the impact of "lid" structure motion on its catalytic performance; from the perspective of compactness of MOFs-enzyme binding, clarify the impact of MOFs on the "lid" structure motion, reveal the internal mechanism of changes of lipase@MOFs catalytic activity; from the perspective of microenvironmental changes of immobilized enzyme, explore microenvironmental changes of lipase@MOFs caused by differences in the "lid" structure motion, identify the molecular interaction mechanism of in-situ encapsulation of lipase in MOFs. The present project will provide theoretical basis and scientific reference for the research of immobilized enzyme catalytic mechanism and enzymology.