Use of engineered enzymes to identify organophosphate and pyrethroid-related toxicity in toxicity identification evaluations

Engineered variants of a carboxylesterase from Lucilia cuprina (E3) and a phosphotriesterase from Agrobacterium radiobacter (OpdA) with enhanced hydrolytic activities against pyrethroid and organophosphate pesticides were evaluated asatoxicity identification evaluation (TIE) manipulation. Reduction in toxicity in the presence of the enzyme provides an indication that the toxicant is the enzyme's target substrate. Carboxy/esterase E3 variants were evaluated to determine if the enzymes could mitigate toxicity of pyrethroids to the amphipod, Hyalella azteca. Enzymes were able to achieve 12-70-fold reduction in toxicity for bifenthrin, cyfluthrin, and cypermethrin in water. Only a 2-fold reduction in toxicity was observed with pyrethroid-contaminated sediment, though the phosphotriesterase OpdA achieved at least a 35-fold reduction in toxicity from the organophosphate chlorpyrifos in sediment. Tests with urban runoff samples and agriculture-affected sediments demonstrated that the enzymes could be useful in TIEs to identify pesticide-related toxicity. The approach promises to be a useful TIE tool for organophosphate and pyrethroid pesticides, particularly in a water matrix, and potentially could be used for identification of toxicity attributable to other pesticides.