Abstract

Enzymatic fuel cells (EFCs) are an emerging class of electrochemical devices that have the potential to harness renewable energy sources by converting the chemical energy of an organic substrate into electrical energy. Enzymes are responsible in an EFC for highly selective and efficient electrode processes under typically mild ambient conditions, such as the oxidation reaction at the anode and the reduction reaction at the cathode 1. In addition to more conventional techniques, electrochemical impedance spectroscopy (EIS) can be employed to characterize such systems. Electrochemical behavior of enzymatic electrodes, such as enzyme reaction kinetics, substrate mass transport, and electrode stability, can be studied by measuring the system's impedance over a wide frequency range and then performing a precise post-process analysis based on the distribution of relaxation time (DRT). DRT analysis is rarely used to interpret EIS spectra within the EFCs research field, despite the fact that it is widely acknowledged as relevant for properly understanding the EIS potential to improve power density in fuel cells 2,3. Glucose oxidase (GOx) and bilirubin oxidase (BOD) were immobilized on multi-walled carbon nanotubes (MWCNTs) to catalyze the bioelectrocatalytic oxidation of glucose at the anode and the bioelectrocatalytic reduction of oxygen at the cathode, respectively. This study presents a promising approach for the investigation of enzyme-based anodic and cathodic processes by measuring them with EIS and then interpreting the results with DRT analysis, as depicted in Figure 1. By varying different operating parameters, such as the enzymatic loading on the electrode surface or electrolyte conditions, this method can be used to identify three primary relaxation processes occurring at both electrodes. At the interface between the electrolyte and the electrode, it is possible to conclude that high-frequency processes are accountable for ionic conduction, intermediate-frequency processes are accountable for charge transfer, and low-frequency processes are accountable for oxygen surface exchange and diffusion.This method provides access to preliminary fundamental information about enzymatic bioelectrode processes, which can be used to optimize biofuel cell construction and compositions and increase their power output. Figure 1. Schematic representation of the characterization method used to characterize a GOx-CNT-modified anode consisting of EIS measurements and subsequent DRT analysis. Concentration of glucose 175 mM in 0.1 M HEPES Buffer, pH 7; EIS parameters: 20 mV sinusoidal excitation, 0.1 Hz-100 kHz. References K. Herkendell, A. Stemmer and R. Tel-Vered, Nano Res., 12(4), 767–775 (2019). H. Wang, X. Long, Y. Sun, D. Wang, Z. Wang, H. Meng, C. Jiang, W. Dong and N. Lu, Frontiers in microbiology, 13, 973501 (2022). F. Ciucci, Current Opinion in Electrochemistry, 13, 132–139 (2019). Figure 1

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