This paper presents a comprehensive study of different chemical pretreatments and enzymatic hydrolysis processes for the production of sugars from lignocellulosic biomass: switchgrass (Panicum virgatum L.) and corn stover. Experimental results were used to scale up the production process for biobutanol. With regards to pretreatments, acid, alkali, surfactants, and organic solvents were tested to break down biomass structure. Acid pretreatment turned out to be the most efficient in terms of total released sugar. Besides, the optimal operating conditions for the enzymatic hydrolysis were determined experimentally. Comparing the degradation of biomass types, switchgrass showed high degradation when sulfuric acid treatment is used. However, corn stover showed the highest sugar production at complete degradation. Therefore, it Will be the preferred raw material and determines the' operating conditions of choice for process scale up.