Biobutanol is an important alternative fuel source, but current processing methods suffer from low yields. This study aimed to develop and optimize a pretreatment protocol for biobutanol production from cornstalks. Fractionation of cornstalks into carbohydrate (cellulose and hemicellulose) and lignin was performed by alkali-catalyzed organosolv pretreatment (ACOS). After optimization of the process parameters, more than 80% of the total lignin was removed, with minimal hemicellulose degradation, at 110 °C, 4% (w/w dry cornstalk) NaOH, 90 min reaction time, and 60% (v/v) ethanol. After enzymatic hydrolysis, the maximum recovery of total monosaccharide was 83.7% (85.0% cellulose, 82.0% hemicellulose). In acetone-butanol-ethanol (ABE) fermentation, a slightly higher total ABE concentration (12.8 g/L vs. 11.9 g/L) was produced from the enzymatic hydrolysate, compared with that from a glucose control. The physical structure and chemical properties of alkali-catalyzed organosolv lignin (ACOSL) showed higher phenolic group content and antioxidant capacity compared with alkali lignin. ACOS pretreatment is an economical method for the production of fermentable monosaccharide and high-value lignin, for use in biofuel production.