Biodetoxification of Organophosphorus Nerve Agents by Immobilized Escherichia coli with Surface Expressed Organophosphorus Hydrolase and CBD Protein in a Fixed-film Bioreactor

本项目旨在通过在固定化细胞生物反应器中将有机磷水解酶（OPH）和纤维素结合域蛋白（CPD）应用于细胞表面的“活”生物催化剂，开发一种生物降解有机磷神经毒剂的新方法，并证明该技术用于神经毒剂大规模解毒的可行性。具体目标是：1)说明OPH和CPD在细胞表面的共表达使活性生物催化剂与纤维素材料紧密结合；2)确定表面显示OPH和CPD的最佳比例，用于解毒和固定化；3)选择合适的纤维素固定化基质，将大量表达OPH的细胞紧密固定，并通过固定化细胞生物载体进行流动；4)以优化的“活”生物催化剂和选定的纤维素固定化载体为基础，开发间歇式和连续式固定化细胞生物反应器，用于有机磷神经毒剂的解毒。＊＊＊

9731513 Mulchandani This project is to develop a novel approach for the biodegradation of organophosphorous nerve agents based on the application of "live" biocatalysts with organophosphorous hydrolase (OPH) and cellulose binding domain protein (CPD) displayed onto the cell surface in immobilized cell bioreactors, and to demonstrate the feasibility of using this technology for large-scale detoxification of nerve agents. Specific objectives are to: 1) Illustrate that the coexpression of OPH and CPD onto the cell surface affords very tight binding of the live biocatalysts to cellulosic materials; 2) Determine the optimal ratio of surface-displayed OPH and CPD for detoxification and immobilization; 3) Select the appropriate cellulose immobiliztion matrix for immobilizing large numbers of OPH expressing cells tightly and for use in a flow through immobilized cell biorector; and 4) Develop batch and continuous immobilized cell bioreactorsbaed on the optimized "live" biocatalysts and selected cellulose immobilization supports for detoxification of organophosphate nerve agents. ***