An Enzyme-Based Biosensor for the Detection of Organophosphate Compounds Using Mutant Phosphotriesterase Immobilized onto Reduced Graphene Oxide

Abstract

Enzymatic detection of organophosphate (OP) compounds can be tailored using highly sensitive and selective enzymes in the development of biosensors. Previously, mutant (YT) phosphotriesterase (PTE) was reported to efficiently hydrolyze Sp and Rp enantiomers of phosphotriester. This study reports the use of phosphotriesterase mutant YT (YT-PTE) immobilized onto reduced graphene oxide (rGO) and fabricated onto a screen-printed carbon electrode (SPCE) for electrochemical detection of OP compounds. Immobilization of YT-PTE onto rGO was secured using N-hydroxysuccinimide (NHS) and N-(3-dimethylaminopropyl)-N’-ethylcarbodiimide (EDC) cross-linker, and the resulting immobilized enzyme was able to retain up to 90% of its activity. Electrochemical analysis of the SPCE/rGO/YT-PTE showed detection of paraoxon in a linear range of 1 mM–0.005 μM with its limit of detection as low as 0.11 μM. SPCE/rGO/YT-PTE exhibited high selectivity towards paraoxon and parathion and have good reproducibility. Furthermore, detection of paraoxon was also possible in a real water sample with only minor interferences.