Engineering bio-interfaces for the direct electron transfer of Myriococcum thermophilum cellobiose dehydrogenase: Towards a mediator-less biosupercapacitor/biofuel cell hybrid

Direct electron transfer (DET) of enzymes on electrode surfaces is highly desirable both for fundamental mechanistic studies and to achieve membrane-and mediator-less bioenergy harvesting. In this report, we describe the preparation and comprehensive structural and electrochemical characterization of a three-dimensional (3D) graphene-based carbon electrode, onto which the two-domain redox enzyme Myriococcum thermophilum cellobiose dehydrogenase (MtCDH) is immobilized. The electrode is prepared by an entirely novel method, which combines in a single step electrochemical reduction of graphene oxide (GO) and simultaneous electrodeposition of positively charged polyethylenimine (PEI), resulting in a well dispersed MtCDH surface. The resulting MtCDH bio-interface was characterized structurally in detail, optimized, and found to exhibit a DET maximum current density of 7.7 +/- 0.9 mu A cm(-2) and a half-lifetime of 48 h for glucose oxidation, attributed to favorable MtCDH surface orientation. A dual, entirely DET-based enzymatic biofuel cell (EBFC) was constructed with a MtCDH bioanode and a Myrothecium verrucaria bilirubin oxidase (MvBOD) biocathode. The EBFC delivers a maximum power density (Pmax) of 7.6 +/- 1.3 mu W cm-2, an open-circuit voltage (OCV) of 0.60 V, and an operational lifetime over seven days, which exceeds most reported CDH based DET-type EBFCs. A bio-supercapacitor/EBFC hybrid was also constructed and found to register maximum power densities 62 and 43 times higher than single glucose/air and lactose/air EBFCs, respectively. This hybrid also shows excellent operational stability with self-charging/discharging over at least 500 cycles.

Automated summary

A novel three-dimensional graphene-based carbon electrode has been successfully utilized for immobilizing Myriococcum thermophilum cellobiose dehydrogenase. Detailed structural and electrochemical characterizations reveal favorable performance in glucose oxidation reactions. The constructed entirely DET-based enzymatic biofuel cell demonstrates excellent power output and reliability.