Comparative in-silico analysis of enzymatic azo dye degradation using laccase, tyrosinase, and peroxidase from Grifola frondosa

Azo dyes, widely used in the textile industry, are toxic, persistent, and resistant to natural degradation, causing significant water pollution and disrupting aquatic ecosystems. This study investigates the enzymatic biodegradation of these pollutants using laccase, tyrosinase, and peroxidase MSP1 from Grifola frondosa . These enzymes, known for their broad substrate specificity, can catalyze the breakdown of complex aromatic structures, making them promising candidates for eco-friendly azo dye removal. Protein sequences were retrieved from NCBI, and physicochemical analysis confirmed structural stability (instability indices <40). Molecular docking with 14 azo dyes identified Congo red as the best-binding substrate, with docking scores of −10.1 (laccase), −10.6 (MSP1), and −11.3 (tyrosinase). Interaction analysis revealed hydrogen bonds and van der Waals forces (1.75–5.42 Å) supporting effective binding. Protein–protein interaction scores were high (0.82, 0.88, and 0.84), and STRING co-expression analysis indicated gene clusters involved in azo dye metabolism. These results suggest that combining these enzymes could synergistically enhance azo dye degradation. Future work should focus on enzyme engineering, experimental validation, and scale-up for industrial wastewater treatment.