The effective deconstruction of lignocellulosic biomass is essential for sustainable biorefineries. In this study, corn stover was pretreated by low-alkali (1–5 wt% NaOH) pre-impregnation assisted instant catapult steam explosion (ICSE) to investigate its influence on enzymatic hydrolysis efficiency and the mechanism of lignin-derived anti-enzymatic factors. The results showed that this pretreatment effectively enhanced glucose yield. Under 4–5% NaOH conditions, washed samples achieved glucose yields above 98%. At 4% NaOH, the glucose yields of washed and unwashed groups were 98.88% and 56.34%, respectively, indicating that washing removed soluble inhibitors. LC-MS analysis identified three major water-soluble inhibitory compounds-vanillin, syringaldehyde, and 2-carboxybenzaldehyde-confirming their negative effects on cellulase activity. The alkali-soluble lignin content of unwashed samples (43.28%) was 1.36 times higher than that of washed samples (31.93%), demonstrating its role as a water-insoluble inhibitory factor. Moreover, SEM, XRD, FTIR, and contact angle analyses revealed that 5% NaOH treatment enhanced lignin solubilization, induced structural rearrangement and interfacial hydrophilic reconstruction, and increased cellulose crystallinity and enzyme accessibility. These findings elucidate the mechanistic pathways of lignin transformation and inhibition mitigation, providing valuable insights for efficient and sustainable biomass conversion.

The effective deconstruction of lignocellulosic biomass is essential for sustainable biorefineries. In this study, corn stover was pretreated by low-alkali (1-5 wt% NaOH) pre-impregnation assisted instant catapult steam explosion (ICSE) to investigate its influence on enzymatic hydrolysis efficiency and the mechanism of lignin-derived anti-enzymatic factors. The results showed that this pretreatment effectively enhanced glucose yield. Under 4–5% NaOH conditions, washed samples achieved glucose yields above 98%. At 4% NaOH, the glucose yields of washed and unwashed groups were 98.88% and 56.34%, respectively, indicating that washing removed soluble inhibitors. LC-MS analysis identified three major water-soluble inhibitory compounds-vanillin, syringaldehyde, and 2-carboxybenzaldehyde-confirming their negative effects on cellulase activity. The alkali-soluble lignin content of unwashed samples (43.28%) was 1.36 times higher than that of washed samples (31.93%), demonstrating its role as a water-insoluble inhibitory factor. Moreover, SEM, XRD, FTIR, and contact angle analyses revealed that 5% NaOH treatment enhanced lignin solubilization, induced structural rearrangement and interfacial hydrophilic reconstruction, and increased cellulose crystallinity and enzyme accessibility. These findings elucidate the mechanistic pathways of lignin transformation and inhibition mitigation, providing valuable insights for efficient and sustainable biomass conversion.