In this study, ethanol organosolv treatment of oat husk and the potential effects of phosphoric acid and oxalic acid as alternatives to sulfuric acid were investigated. These acids were determined as effective as sulfuric acid to obtain high quality lignin and glucan and they can be used instead of sulfuric acid in solvent acidification. To determine the purity and recovery of both lignin and glucan, the effects of initial substrate amount, solid-to-liquid ratio, and amount of washing solutions were also examined using a one-factor-at-a-time strategy. Reducing the amount of washing solutions (water, solvent, or both) negatively affected lignin recovery, but it did not affect glucan recovery. The optimum conditions for pretreatment of the oat husk at higher glucan recovery were obtained with 50% aqueous ethanol acidified with oxalic acid at 210 °C for 90 min and solid-to-liquid ratio of 1:2. In the mixture of evaporated glucan-rich and hemicellulose-rich fractions obtained through the optimized condition, 4.62 g/L biomass containing 10.27% protein was produced by the cultivation of Aspergillus oryzae. The fractions obtained from organosolv treatment can be used to obtain value-added products such as biomass production, and thus contributing to a sustainable economy by integrating lignocellulosic substrate residues into the biorefinery.