基于漆酶逆胶束体系的含油污泥中多环芳烃降解的方法与机理研究

The existence of polycyclic aromatic hydrocarbons in the oily sludge poses a serious threat to the surrounding soil, water and human being. It is known that microbial enzymes possess the ability to degrade some environmental pollutants in municipal sludge. However, those enzymes will fade away in terms of oily sludge due to its contents. This project will construct reverse micelles by selecting biosurfactant in the extracted oil from oily sludge, in order to study the catalytic behavior of laccase and degradation of polycyclic aromatic hydrocarbons in the oily sludge. Numbers of modern technologies such as circular dichroism spectrum, nuclear magnetic resonance, electron spin resonance, fourier transform infrared spectroscopy will be applied to investigate the structure, activity and conformational change of laccase. This project aims at inducing the activated conformation of both laccase and polycyclic aromatic hydrocarbons, locating the binding site of laccase and polycyclic aromatic hydrocarbons, definitizing the effect of conformation change on degradation efficiency. Also, this project is supposed to unveil the mechanism of degradation of polycyclic aromatic hydrocarbons in reverse micelle system. Meanwhile, degradation kinetic models of laccase and polycyclic aromatic hydrocarbons are supposed to be built up. This project focuses on the new method in degradation of environment pollutants polycyclic aromatic hydrocarbons in oily sludge, and guiding the theoretical basis in control of pollution in environment.

含油污泥中多环芳烃的存在对周围土壤、水体和人类构成严重的威胁。微生物酶对市政污泥中污染物具有降解作用，然而对于含油污泥，酶与含油污泥直接接触导致酶活性和稳定性受到影响，一定程度上制约着含油污泥中多环芳烃的降解。本项目选择生物表面活性剂，提取含油污泥的油分并构建逆胶束体系，研究漆酶在油分-逆胶束中活性、催化性能等变化，并研究多环芳烃的降解行为。项目拟采用圆二色谱、核磁共振、电子自旋共振、傅里叶变换红外光谱等技术深入研究油分-逆胶束体系中漆酶与多环芳烃的空间结构和构象变化，在油分-逆胶束体系中诱导出漆酶和多环芳烃的活性构象，探究漆酶与多环芳烃的结合位点，并探讨油分-逆胶束中多环芳烃在漆酶作用下降解的途径与作用机制，建立漆酶构象、多环芳烃构象对降解多环芳烃影响的动力学模型。项目成果将为含油污泥中多环芳烃的降解提供方法，对降解环境污染物多环芳烃的高效绿色方法提供理论支持和技术途径。

本项目围绕漆酶逆胶束体系降解多环芳烃，开展了基于漆酶逆胶束体系的含油污泥中多环芳烃降解的方法与机理研究。主要内容包括：考察并对比了漆酶逆胶束与非漆酶逆胶束体系在不同外界条件下漆酶对多环芳烃的催化降解的影响，确定了漆酶在逆胶束体系下催化降解多环芳烃的最佳条件；通过圆二色谱以及红外光谱等相关分析技术，系统地比较漆酶逆胶束体系与非漆酶逆胶束体系在不同条件下漆酶二级结构的变化，探明了漆酶二级结构中各结构占比与漆酶活性的关系；利用气相质谱联用仪对降解过程中的中间产物进行检测，并通过对Hishfeld电荷和福井函数进行相关量子化学的计算，预测了漆酶催化多环芳烃的初始降解位点等。本项目从理论和实践上支持并完善了漆酶在逆胶束体系下降解多环芳烃的降解机理，为降解环境污染物多环芳烃高效的绿色方法提供了理论支持和技术途径。