Obtaining value-added products from renewable resources is limited by the lack of specific operating conditions optimized for the physico-chemical characteristics of the biomass and the desired end product. A mathematical model and statistical optimization were developed for the production of volatile fatty acids (VFAs) by anaerobic digestion of municipal sewage sludge. The experimental tests were carried out in triplicate and investigated a wide range of conditions: pH 9.5, 10.5, and 11.5; temperatures 25 °C, 35 °C, 45 °C, and 55 °C; primary sludge with organic loading (OL) of 10 and 14 g VS (volatile solids); and digested sludge with 4 and 6 g VS. Subsequently, a statistical search was performed to obtain optimal production conditions, then a statistical model of VFA production was developed and the optimal conditions were validated at pilot plant scale. The maximum VFA concentration predicted was 6975 mg COD (chemical oxygen demand)/L using primary sludge at 25 °C, initial OL of 14 g VS, and pH 10.5. The obtained third-degree model ( r ² = 0.83) is a powerful tool for bioprocess scale-up, offering a promising avenue for sustainable waste management and biorefinery development.