Abstract
Bitter taste substances commonly represent a signal of toxicity. Fast and reliable detection of bitter molecules improves the safety of foods and beverages. Here, we report a biosensor using an easily accessible and cost-effective odorant-binding protein (OBP) of Drosophila melanogaster as a biosensitive material for the detection of bitter molecules. Based on the theoretical evaluation of the protein-ligand interaction, binding energies between the OBP and bitter molecules were calculated via molecular docking for the prediction and verification of binding affinities. Through one-step reduction, gold nanoparticles (AuNPs) and reduced graphene oxide (rGO) were deposited on the screen-printed electrodes for improving the electrochemical properties of electrodes. After the electrodes were immobilized with OBPs via layer-by-layer self-assembly, typical bitter molecules, such as denatonium, quinine, and berberine, were investigated through electrochemical impedance spectroscopy. The bitter molecules showed significant binding properties to the OBP with linear response concentrations ranging from 10-9 to 10-6 mg/mL. Therefore, the OBP-based biosensor offered powerful analytic techniques for the detection of bitter molecules and showed promising applications in the field of bitter taste evaluation.