河流湖泊底泥中积累的大量难降解有机物对水生态系统构成长期威胁。向底泥中添加固定化酶可以高效降解某些难降解有机物，启动或强化微生物降解进程。然而，固定化酶的构象变化及其与酶活性、与底泥底物交互作用的内在关联机制仍未得到阐明，一定程度上制约了固定化酶在底泥难降解有机物污染修复中的应用及其催化降解效率的提高。本项目选择变孔径介孔碳为代表性载体，固定化多种功能酶，用于河湖底泥酚类难降解有机物污染修复。拟运用激光共聚焦显微镜、高分辨电镜、拉曼光谱、核磁共振、时间分辨荧光光谱、原子力显微镜等技术结合分子动力学模拟来深入研究固定化酶的空间分布及团聚、活性及构象变化以及与底泥底物的交互作用机制，并探究表面活性剂对污染修复过程的影响，从微观尺度阐明固定化酶应用于河湖底泥难降解有机物污染原位修复的作用机制及环境影响。项目成果将为深入研究底泥难降解有机物污染原位修复、有效控制河湖底泥污染提供理论依据和技术支撑。

Numbers of recalcitrant organics accumulated in the sediment of rivers and lakes pose a long term threat to the aqueous ecological system. The addition of immobilized enzymes into the sediment can efficiently degrade some recalcitrant organics, which will initiate or enhance the microbial degradation process. However, the conformational change of immobilized enzymes and its internal correlation with enzyme activity and interaction with sediment substrate of the immobilized enzyme are still unclear, hindering the application of immobilized enzymes in the remediation of sediment recalcitrant organics contamination and the promotion of their catalytic degradation ability. This project will select the mesoporous carbon material with adjustable pore size as the representing carrier for immobilizing several functional enzymes to be applied in the remediation of sediment contamination by phenolic recalcitrant organics in rivers and lakes. Confocal laser scanning microscope, high-resolution electron microscope, Raman spectrometry, nuclear magnetic resonance, time-resolved fluorescence spectrometry, atomic force microscope as well as molecular dynamics simulation will be used to profoundly study the spatial distribution, aggregation, activity and conformational change of the immobilized enzymes, and their interaction mechanism with the sediment substrate. The effect of surfactants on the remediation process will also be explored. The reaction mechanisms and the environmental impact of the immobilized enzyme for in-situ remediation of sediment recalcitrant organics contamination in rivers and lakes will be revealed on the micro-scale level. The research results of this project will provide theoretical evidence and technical support for profound study of in-situ remediation of sediment recalcitrant organics contamination and effective control of sediment pollution in rivers and lakes.