Wire-ready invertase-glucose dehydrogenase chimera as a strategic design for enzymatic chain reaction-driven electrocatalysis

Abstract

The multienzymatic reaction in bioelectrocatalytic systems can offer promising pathways for diverse biochemical synthesis or higher electricity generation. To achieve efficient enzymatic chain reactions and electron transfer at enzyme-electrode interfaces, precise control of enzyme positioning and surface-orientation is paramount. Therefore, the selection of multienzyme co-immobilization techniques must be approached with great care. In this study, we describe a “solid binding peptide (SBP)-fused chimeric enzyme” aimed at directly knotting the invertase (INV), glucose dehydrogenase gamma-alpha complex (GDHγα), and electrode, sequentially. The INV and GDHγα, positioned upstream and downstream enzymes, were linked via a flexible oligopeptide linker. Subsequently, GDHγα, acting as a redox protein responsible for electron production, was immobilized on electrode using an SBP tethered to the GDH α subunit. We show that the “wire-ready fusion construct” significantly enhanced oxidative currents on the electrode compared to the cases where either inter-enzyme proximity or cofactor-electrode distance is controlled individually. © 2024