Strategies for Metabolic Engineering of Environmental Microorganisms-Application to Degradation of Organophosphate Contaminants

The goal of this work was to develop the experimental and theoretical methods to introduce multiple, heterologous, biodegradation pathways into a single organism and to optimize the flux through those pathways for the remediation of toxic or recalcitrant organic contaminants. Specifically, we found and cloned a gene that encodes an enzyme capable of degrading diethylphosphate, cloned and expressed a pathway for complete mineralization of p-nitrophenol phosphate, cloned and expressed a phosphotriesterase capable of hydrolyzing parathion, developed a co-culture biofilm capable of degrading ding parathion (as a proof-of-concept), and combined all of the genes in a single organism for complete mineralization of parathion of paraoxon.