Dilute-acid pretreatment has proven to be a robust means of converting herbaceous feedstock to fermentable sugars. However, it also releases acetic acid, a known fermentation inhibitor, from acetyl groups present in the biomass. A mild, dilute alkaline extraction stage was implemented prior to acid pretreatment to separate acetic acid from the hydrolysate sugar stream. This step, termed deacetylation, improved the glucose and xylose yields from enzymatic hydrolysis and ethanol yields from fermentation of the sugars relative to the control experiments using dilute-acid pretreatment of native corn stover without deacetylation. While promising, deacetylation as it was historically practiced is conducted at low solids loadings, and at fixed conditions. Thus, many questions have been left unanswered, including the relationship between sodium hydroxide and solids loading, and acetate and xylan solubilization, as well as the impact of temperature and residence time on the process efficacy. A central composite experiment was designed to evaluate the impact of solids loading, sodium hydroxide loading, reaction time and temperature during deacetylation on the acetate and xylan solubilization of corn stover. Using the ANOVA test, it became apparent that neither of the responses was significantly impacted by the solids loading, while the reaction time was a minor factor - the responses were largely driven by reaction temperature and the sodium hydroxide loading. Based on the results, we successfully demonstrated the ability to transition the low-solids (10 % w/w) deacetylation process to a higher-solids process (30 % w/w) with minimal impact on the ability to extract acetate from biomass. Conditions were selected to minimize xylose loss during deacetylation, while also removing 70 % of acetyl groups. The impact of selected conditions on the enzymatic hydrolysis and fermentation was further investigated. In conclusion, evaluation of the whole-process impact demonstrated that despite the upfront reduction in carbohydrate loss during deacetylation, the overall process sugar yields were depressed by the high-solids, low alkali process relative to the historical control. Consequently, ethanol titers were reduced, though strong fermentation performance was still observed, indicating that 70 % acetate removal is sufficient to depress acetic acid concentrations to a level that does not substantially inhibit ethanol fermentation by rZymomo nas.