Evaluation of Enzymatic Hydrolysis of Sugarcane Bagasse Using Combination of Enzymes or Co-Substrate to Boost Lytic Polysaccharide Monooxygenases Action

Abstract

This study evaluated innovative approaches for the enzymatic hydrolysis of lignocellulosic biomass. More specifically, assays were performed to evaluate the supplementation of the commercial cellulolytic cocktail Cellic® CTec2 (CC2) with LPMO (GcLPMO9B), H2O2, or cello-oligosaccharide dehydrogenase (CelDH) FgCelDH7C in order to boost the LPMO action and improve the saccharification efficiency of biomass into monosaccharides. The enzymatic hydrolysis was carried out using sugarcane bagasse pretreated by hydrodynamic cavitation-assisted oxidative process, 10% (w/w) solid loading, and 30 FPU CC2/g dry biomass. The results were compared in terms of sugars release and revealed an important influence of the supplementations at the initial 6 h of hydrolysis. While the addition of CelDH led to a steady increase in glucose production to reach 101.1 mg of glucose/g DM, accounting for the highest value achieved after 72 h of hydrolysis, boosting the LPMOs activity by the supplementation of pure LPMOs or the LPMO co-substrate H2O2 were also effective to improve the cellulose conversion, increasing the initial reaction rate of the hydrolysis. These results revealed that LPMOs play an important role on enzymatic hydrolysis of cellulose and boosting their action can help to improve the reaction rate and increase the hydrolysis yield. LPMOs-CelDH oxidative pairs represent a novel potent combination for use in the enzymatic hydrolysis of lignocellulose biomass. Finally, the strategies presented in this study are promising approaches for application in lignocellulosic biorefineries, especially using sugarcane bagasse as a feedstock.