AbstractCoastal Bermuda grass (CBG) is an agricultural residue with considerable potential as a feedstock for lignocellulosic-based ethanol. The treatment of biomass with water at high temperature, termed autohydrolysis, can be used to recover sugars in the filtrate and to improve enzyme digestibility of the pretreated solids. The effect of a two- stage autohydrolysis process with respect to total sugar recovery relative to a one stage process was investigated. CBG was subjected to lab scale one-stage (150, 160, and 170 °C) and two-stage (150/170 °C and 160/170 °C) isothermal autohydrolysis processes followed by enzyme hydrolysis on the residual solids with different loadings (5 to 30 FPU/g). Two-stage autohydrolysis (160/170 °C) solubilized 94.2% of the hemicellulose based on the original CBG material but only 17.7% of the cellulose and 30.4% of the lignin. Increases in the severity factor (a combination of time and temperature) of autohydrolysis pretreatments decreased the recoverable carbohydrates and total solids. Two-stage autohydrolysis enhanced enzyme digestibility of the cellulose in pretreated solids relative to one-stage autohydrolysis, especially at higher values of FPU/g. The overall total theoretical sugar recovery achievable by the two stage process was 57.8% and for the one stage process only 51.6% with 30 FPU/g. This marginal increase would have to be considered relative to increased complexity of operations when deciding whether to implement one or two stage autohydrolysis.