Development of a Fluorescence-based Biosensor for Coumaphos

Abstract

A screening procedure has been implemented based on the inverse virtual screening technique to identify new bioreceptors for the set-up of pesticide biosensors. Different pesticides of interest were docked on a limited database of proteins with known 3D structures. Some compounds of interest were chosen among the following classes: organophosphates, organo-clorurates, carbamates, anilides, neonicotinoids, aromatic and heterocyclic azoto-organics. Hundreds of molecules were automatically docked on hundreds of structures and results analysed. We identified many interesting binders among which we focused on an E. coli helicase that was demonstrated to be able to bind the organophosphate Coumaphos on multiple sites. In addition we report on the purification of the binder and testing with Coumaphos and other compounds in a fluorescence-based assay.

Keywords

• helicase
• pesticides
• docking analysis
• bioreceptor
• biosensor
• fluorescence-based assay