Simultaneous enhancement of activity and stability of Bacillus safensis-derived laccase and its application in lignocellulose saccharification.

Effective hydrolysis of lignocelluloses for producing reducing sugar is impeded by the covalent binding of hemicellulose and cellulose through lignin, which could be eliminated by laccases. This study identified a novel thermostable laccase from Bacillus safensis TCCC 111022 and created an iterative mutant E231D/Y441H, exhibiting 1.59-fold greater specific activity and a 183 % greater half-life at 80°C than the wild-type enzyme. Computational analysis revealed that the stability and activity of the E231D/Y441H could be simultaneously enhanced by increasing the flexibility of the ring around the substrate binding pocket. Additionally, the saccharification efficiency of sugarcane bagasse and corn stalks were both enhanced by 235 % in the system adding E231D/Y441H, mixed-cellulases, and mediator (1-hydroxybenzotriazole) compared to the samples treated with mixed-cellulases. The findings of this research provide a reference for the degradation of lignocellulosic substrates and contribute to the sustainable development of biomass-based industries.