PHP-Family Diesterase from with Broad Specificity and High Catalytic Efficiency against Organophosphate Flame-Retardant Derived Diesters.

Organophosphate flame retardants have been widely used in plastic products since the early 2000s. Unfortunately, these compounds leach out of the plastics over time and are carcinogenic, developmental toxins, and endocrine disruptors. Due to the high usage levels and stable nature of the compounds, widespread contamination of the environment has now been observed. Despite their recent introduction into the environment, bacteria from the family have evolved a three-step hydrolytic pathway to utilize these compounds. The second step in this pathway in sp. TCM1 is catalyzed by -PDE, which is a member of the polymerase and histidinol phosphatase (PHP) family of phosphatases. This enzyme is only the second case of a PHP-family enzyme capable of hydrolyzing phosphodiesters. Bioinformatics analysis has now been used to identify a second PHP diesterase from sp. EMRT-2 (-PDE). Kinetic characterization of -PDE and -PDE with authentic organophosphate flame-retardant diesters demonstrates that these enzymes are true diesterases with more than 1000-fold selectivity for the diesterase activity seen in some cases. Synthesis of a wide array of authentic flame-retardant diesters has allowed the substrate specificity of these enzymes to be determined, and mutagenic analysis of the active site residues has identified key residues that give rise to the high levels of diesterase activity. Despite high sequence identity, -PDE is found to have a broader substrate specificity against flame-retardant derived diesters, and / values greater than 10 M s are seen with the best substrates.