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

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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.