Exploring how lignin structure influences the interaction between carbohydrate-binding module and lignin using AFM

Nonproductive adsorption of cellulase onto the residual lignin in substrate seriously hinders the enzymatic hy-drolysis. To understand how lignin structure affects lignin-cellulase interaction, the carbohydrate-binding module (CBM) functionalized atomic force microscope tip was used to measure CBM-lignin interaction by single-molecule dynamic force spectroscopy in this work. The results showed that sulfonated lignin (SL) has the greatest adhesion force to CBM (4.74 nN), while those of masson pine milled wood lignin (MWL), poplar MWL and herbaceous MWLs were 2.85, 1.03 and 0.27-0.61 nN, respectively. It provides direct quantitative evidence for the significance of lignin structure on lignin-cellulase interaction. The CBM-MWLs interaction decreased sharply to 0.054-0.083 nN while SL was added, indicating the primary mechanism of SL promoting lignocel-lulose hydrolysis was significantly reducing the nonproductive adsorption of substrate lignin on cellulase. Finally, the "competitive adsorption" mechanism was proposed to interpret why SL effectively promotes the enzymatic hydrolysis of lignin-containing substrates.