Improving the specificity of organophosphorus hydrolase to acephate by mutagenesis at its binding site: a computational study.

Organophosphorus hydrolase (OPH) is one of the most important enzymes in order to bioremediation of organophosphorus (OP) pesticides. OPH is capable of degrading a wide variety of OPs, but it has poor specificity to OPs with P-S bond, including acephate. Given that the binding site residues of OPH determine its substrate specificity, this study was carried out to find the best OPH mutants containing a single point mutation in the binding site that possess improved specificity to acephate. Hence, we generated all possible mutant models and performed molecular docking of acephate with wild-type OPH (OPH-WT) and its mutants. After that, molecular dynamic (MD) simulations of OPH-WT and the best mutants, according to the docking results, were performed in both apo- and complex with acephate forms. Docking results signified that 51 out of 228 mutants possessed increased binding affinities to acephate, as compared to OPH-WT. Of them, W131N, W131G, and H254Y were the best mutants considering the high binding affinities and proper orientation of the ligand at their active sites. MD simulations confirmed the stability of the three mutants in both apo- and complex with acephate forms and also showed that these formed more stable complexes with acephate, as compared to OPH-WT. MD results also suggested that W131N and W131G, in addition to enhanced specificity, could keep the necessary configuration for acephate hydrolysis during the simulations.