Construction of novel cellulase with high activity, great stability and excellent lignin-resistant for efficient enzymatic hydrolysis by hydrophilic and negatively charged modification

The lignin-derived inhibition on cellulase is the most critical bottleneck that must be addressed when converting lignocellulosic biomass through the sugar-platform process. Herein, a novel modified cellulase with high activity, great stability and excellent lignin-resistant ability was synthesized by chemical modification using hydrophilic and negatively charged polyethylene glycol (PEG) derivatives including HO-PEG-COOH and HOOC-PEG-COOH. Both modifications obviously increased the cellulase activity, with 37% (HO-PEG-COOH modification) and 53% (HOOC-PEG-COOH modification) increase, respectively. In addition, the stabilities of both modified cellulases were significantly enhanced, seeing from their much higher activities at all selected temperatures and pH values. The modification resulted in a movement of tryptophan residues to the exterior of enzyme, allowing the substrate chains to be more easily recruited and thus initiating the hydrolysis. The compactness of secondary structure for enzyme was improved, especially, the α-helix content was increased which requires strong hydrogen bonding to generate it, thus enhancing the stability. High hydrophilicity, negative charge and compactness of enzyme conferred modified cellulase good ability to resist lignin adsorption and destruction, thus improving the enzymatic hydrolysis of corn stover. • Cellulase activity is significantly improved by up to 52% after chemical modification. • The migration of Trp residues allows the hydrolysis to be more easily initiated. • High compactness gives modified cellulase great stability and low level of damage by lignin. • High hydrophilicity and negative charge of modified cellulase prevent lignin adsorption. • The novel modified cellulase exhibits promising potential in converting corn stover.