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This research focuses on the rational design of porous enzymatic electrodes, using horseradish peroxidase (HRP) as a model biocatalyst. Our goal was to identify the main obstacles to maximizing biocatalyst utilization within complex porous structures and to assess the impact of various carbon nanomaterials on electrode performance. We evaluated as-synthesized carbon nanomaterials, such as Carbon Aerogel, Coral Carbon, and Carbon Hollow Spheres, against the commercially available Vulcan XC72 carbon nanomaterial. The 3D electrodes were constructed using gelatin as a binder, which was cross-linked with glutaraldehyde. The bioelectrodes were characterized electrochemically in the absence and presence of 3 mM of hydrogen peroxide. The capacitive behavior observed was in accordance with the BET surface area of the materials under study. The catalytic activity towards hydrogen peroxide reduction was partially linked to the capacitive behavior trend in the absence of hydrogen peroxide. Notably, the Coral Carbon electrode demonstrated large capacitive currents but low catalytic currents, an exception to the observed trend. Microscopic analysis of the electrodes indicated suboptimal gelatin distribution in the Coral Carbon electrode. This study also highlighted the challenges in transferring the preparation procedure from one carbon nanomaterial to another, emphasizing the importance of binder quantity, which appears to depend on particle size and quantity and warrants further studies. Under conditions of the present study, Vulcan XC72 with a catalytic current of ca. 300 µA cm−2 in the presence of 3 mM of hydrogen peroxide was found to be the most optimal biocatalyst support.
Varničić et al. (Wed,) studied this question.