Site-directed lysine modification of xylanase for oriented immobilization onto silicon dioxide nanoparticles

Enhanced covalent immobilization of xylanase from Chaetomium globosum (XylCg) onto SiO₂ nanoparticles was achieved by the modification of surface residues. The mutation of surface residues to lysine by site-directed mutagenesis increased the immobilization efficiency (IE) and immobilization yield (IY). The immobilized mutant XylCg (N172K-H173K-S176K-K133A-K148A) exhibited an IY of 99.5% and IE of 135%, which were 1.8- and 4.3-fold higher than immobilized wildtype (WT). Regarding the catalytic properties, the k_cat and k_cat/K_m values were 1850 s^-1 and 2030 mL mg^-1 s^-1 for the immobilized mutant, and 331 s^-1 and 404 mL mg^-1 s^-1 for the immobilized WT, respectively. Additionally, the immobilized mutant exhibited four times higher thermal stability than the immobilized WT at 60 °C. These results suggest that surface-mutated lysine residues confer good stability and orientation on the support matrix, thus improving the overall performance of xylanase.