ME: Redirecting Cellular Metabolism for the Biodegradation of Mixtures of Chlorinated Solvents

该项目的目标是通过代谢工程微生物对四氯乙烯(PCE)、三氯乙烯(TCE)以及这些和其他溶剂的混合物进行快速、长期、好氧的生物降解，从而创建氯化溶剂生物修复的新范例。目前还没有任何生物具有这些能力，开发这种菌株将为解决世界上一些最普遍的土壤和地下水污染问题提供一个新的、具有成本效益的选择。这项研究有四个具体目标：(1)使用DNA改组来创造一个优化的单加氧酶，它可以降解PCE、TCE和氯化溶剂的混合物；(2)将这种单加氧酶和环氧化物水解酶克隆到两个宿主菌株中；(3)使用二维蛋白质电泳评估这种基因工程对宿主细胞生理的影响；以及(4)评估土壤和生物反应器环境中改良细胞和导入基因的命运。

The objective of this project is to create a new paradigm in bioremediation of chlorinated solvents by metabolically engineering microorganisms for the rapid, long-term, aerobic biodegradation of perchloroethylene (PCE), trichloroethylene (TCE), and mixtures of these and other solvents. No organisms are known to have these abilities, and development of such a strain would provide a new, cost-effective option for solving some of the world's most pervasive soil and groundwater pollution problems. This research has four specific objectives: (1) use DNA shuffling to create an optimized monooxygenase that degrades PCE, TCE, and mixtures of chlorinated solvents; (2) clone this monooxygenase and epoxide hydrolase into two host strains; (3) evaluate the effects of this genetic engineering on host cell physiology using 2-dimensional protein electrophoresis; and (4) evaluate the fate of modified cells and introduced genes in soil and bioreactor settings.