Site-specific immobilization of gold binding polypeptide on gold nanoparticle-coated graphene sheet for biosensor application

The effective and strong immobilization of enzymes on solid surfaces is required for current biological applications, such as microchips, biofuel cells, and biosensors. Gold-binding polypeptide (GBP), a genetically designed peptide, possesses unique and specific interactions with a gold surface, resulting in improved enzyme stability and activity. Herein we demonstrated an immobilization method for biosensor applications through site-specific interactions between GBP-fused organophosphorus hydrolase (GBP-OPH) and gold nanoparticle-coated chemically modified graphene (Au-CMG), showing enhanced sensing capability. A flow injection biosensor was fabricated by using GBP-OPH/Au-CMG to detect paraoxons, a model pesticide, showing higher sensitivity, lower detection limit and better operating stability compared that of OPH/Au-CMG. This strategy, which integrates biotic and abiotic moieties through site-specific interactions, has a great potential for use in biosensing and bioconversion process.