AbstractH2O2 is both bactericidal and the main oxidant responsible for lignin degradation reaction catalyzed by manganese peroxidase (MnP) and lignin peroxidase (LiP). Thus, H2O2 treatment of corn stalk and the implementation of solid-substrate fermentation (SSF) is possible to increase the removal rate of lignin from stalk in the process of SSF and after SSF, while avoiding the need to sterilize the raw materials. To demonstrate this approach, SSF was initially carried out using corn stalk pretreated with different concentrations of H2O2 as a substrate. A. oryzae was found to grow well in the 3% H2O2-pretreated corn stalk. H2O2-pretreated corn stalk showed increased MnP and LiP synthesis and disintegration of lignin, but inhibited cellulase synthesis and cellulose degradation. Production of the SSF (200 g) on the 10th day was hydrolyzed in the presence of additional 600 mL different concentration of H2O2 aqueous solution. The total removal of lignin (73.15%) of hydrolysis for 10 h at 3% H2O2 solution was highest and far higher than that at the 12th day, as achieved by conventional SSF. Applying this strategy in practice may shorten the time of lignin degradation, increase the removal of lignin, and decrease the loss of cellulose. Thus, this study has provided a foundation for further study saccharification of corn stalk.