Fractionation of corn stover (CS) was carried out by bisulfite pretreatment in order to improve the production of bioethanol and high-value chemicals. Firstly, the optimum bisulfite pretreatment conditions of CS (170 C, 30 min, 7% NaHSO3, 1% H2SO4) were identified. Next, a biorefinery process of bisulfite pretreatment for CS was proposed. CS was separated into solid and liquor components using such pretreatment. The solid components were employed for bioethanol production by quasi-simultaneous saccharification and fermentation (Q-SSF). The bisulfite liquor was fractionated into hemicellulosic sugars and lignin by different types of resins. It was shown that CS components could be effectively fractionated through bisulfite pretreatment in combination with resin separation to produce bioethanol, hemicellulosic sugars, and lignosulfonate.