The aim of this study was to investigate the changes in enzyme adsorption to solids and cellulose conversion during the hydrolysis of pretreated corn stover with different substrate and enzyme loadings. Enzyme adsorption achieves equilibrium in the early stage and is stable throughout the hydrolysis process. Enzyme adsorption decreases with the increasing loading of enzyme, except for a system with 15% (w/v) substrate. The inhibition on cellulose digestibility brought by higher substrate loading cannot be easily relieved through elevation of enzyme loading. Glucose does not have an inhibition effect on cellulase adsorption to the solids fraction, while the enzyme is unable to transition from partially degraded cellulose to liquid but can be adsorbed again by lignin. The glucose concentration and ratio of cellulose and lignin are two factors that influence enzyme distribution.