Advanced strategies for enzyme–electrode interfacing in bioelectrocatalytic systems

Advances in protein engineering-enabled enzyme immobilization technologies have significantly improved enzyme-electrode wiring in enzymatic electrochemical systems, which harness natural biological machinery to either generate electricity or synthesize biochemicals. In this review, we provide guidelines for designing enzyme-electrodes, focusing on how performance variables change depending on electron transfer (ET) mechanisms. Recent advancements in enzyme immobilization technologies are summarized, highlighting their contributions to extending enzyme-electrode sustainability (up to months), enhancing biosensor sensitivity, improving biofuel cell performance, and setting a new benchmark for turnover frequency in bioelectrocatalysis. We also highlight state-of-the-art protein-engineering approaches that enhance enzyme-electrode interfacing through three key principles: protein-protein, protein-ligand, and protein-inorganic interactions. Finally, we discuss prospective avenues in strategic protein design for real-world applications.