Optimizing Covalent Immobilization of Glucose Oxidase and Laccase on PV15 Fluoropolymer-Based Bioelectrodes

Enzymatic biofuel cells (EBCs) represent a promising technology for biosensors, biodevices, and sustainable green energy applications, thanks to enzymes' high specificity and catalytic efficiency. Nevertheless, drawbacks such as limited output power and short lifetime have to be solved. Nowadays, research is addressed to the use of 3D electrode structures, but the high cost and the industrialization difficulties of such electrodes represent a key issue. The purpose of the paper is thus to describe the use of a low-cost commercial conductive polymer (Sigracell^® PV15) as support for the covalent immobilization of glucose oxidase and laccase, for bioanode and biocathode fabrication, respectively. Efficient immobilization protocols were determined for the immobilized enzymes in terms of employed linkers and enzyme concentrations, resulting in significant enzymatic activities for units of area. The analysis focuses specifically on the optimization of the challenging immobilization of laccase and assessing its stability over time. In particular, an optimum activity of 23 mU/cm² was found by immobilizing 0.18 mg/cm² of laccase, allowing better performances, as for voltage output and electrochemical stability, and a direct electron transfer mechanism to be revealed for the fabricated biocathode. This study thus poses the basis for the viable development of low-cost functional EBC devices for biomedical applications.