Recent advances in laccase: enzyme discovery, molecular modification and application to electrochemical biosensors in nanomaterials

Laccases, as versatile multi-copper oxidases, exhibit broad potential in electrochemical biosensors due to their ability to catalyze redox reactions under mild conditions. However, their widespread application is hindered by limitations in enzyme diversity, stability under extreme conditions (e.g., temperature, pH, ionic strength), and compatibility with nanomaterials. Recent advancements in enzyme discovery, molecular engineering, and nanomaterial development aim to address these challenges. This review synthesizes recent progress in three key areas: (i) discovery of novel laccases from diverse microbial sources (e.g., thermophilic bacteria, psychrophilic fungi) with enhanced stability and substrate specificity; (ii) molecular modification strategies, including rational design, directed evolution, and site-directed mutagenesis, to optimize redox potential, substrate binding, and environmental tolerance; (iii) integration of laccases with advanced nanomaterials (e.g., graphene, carbon nanotubes, metal nanoparticles, MOFs) to improve electron transfer efficiency, enzyme immobilization, and sensor performance. Many new enzymes have given us more confidence to explore more sources of excellent enzyme components for sensors, and the continuous updating and advancement of molecular modification technology has brought us the most important tools to enhance the enzyme properties according to the scenarios and needs of the sensors, and the rapid development of nanomaterials will better enhance the performance of the sensors in terms of detection limit, sensitivity, and the limitation of the scenarios of applications. • Comprehensive Coverage: Presents an up-to-date review of laccase research. • Novel Enzyme Sources: Highlights recent discoveries of extremophilic laccases. • Engineering Strategies: Details cutting-edge molecular modification techniques. • Nanomaterial Integration: Explores innovative approaches for laccase immobilization. • Performance Metrics: Systematically compares key parameters of laccase biosensors.