AbstractThis study investigated the degradation and enzymatic saccharification of cornstalk by white-rot and brown rot fungi. The fungal strains Trametes pubescens Cui 7571, Trametes velutina Dai 10149, and Antrodia wangii Cui 7568 were analyzed in solid-state fermentation cultures. Various extracellular enzyme activities were assessed to determine biochemical changes during the degradation process. Fourier-transform infrared (FTIR) spectroscopy and X-ray diffraction (XRD) analysis were used to determine the structural changes. A correlation analysis between the chemical composition of cornstalk, extracellular enzyme activities, and structural changes indicated that T. pubescens Cui 7571 broke down lignin efficiently and subsequently degraded cellulose, and hemicellulose digestion was not affected by the lignin barrier. Enzymatic hydrolysis demonstrated that the T. pubescens Cui 7571-pretreated samples increased cellulose and hemicellulose conversion in cornstalk. Overall, T. pubescens Cui 7571 displayed excellent performance as a biological pretreatment agent, and lignin played a significant role in the enzymatic saccharification of cornstalk.