The peels of pequi (Caryocar brasiliense Camb.), native to the Brazilian Cerrado, represent ∼80% of the mass of the whole fruit. Despite their high quantities of dietary fiber and bioactive compounds, they are generally discarded as waste. The impact of substituting rice flour with pequi mesocarp flour (PMF) in different concentrations in biscuit characteristics was determined. Pequi mesocarp was converted into flour through drying and milling, and used in quantities of 10%, 20%, and 30% in biscuit formulations. Biscuits were evaluated by their composition, physical parameters, microstructure, and antioxidant capacity. PMF was responsible for increasing protein and ash contents in biscuits. Besides that, higher concentrations of PMF resulted in a darker color of biscuits, higher spread ratio and firmness, and lower weight loss and specific volume. Such effects were attributed to the water and oil-holding capacity because of dietary fibers present in pequi mesocarp. All biscuits produced with PMF showed antioxidant capacity, it increased with higher substitutions. Even though using PMF resulted in darker and firmer biscuits, the results indicate that PMF has the potential to produce functional gluten-free bakery products. PRACTICAL APPLICATION: Pequi peels have a rich composition representing an opportunity to improve the nutritional attributes of bakery products. The pequi mesocarp biscuits produced are gluten-free and contain antioxidants, promoting consumers' health. Producing biscuits with pequi mesocarp is advantageous to offer the population a nutritive alternative biscuit, while adding value to pequi peels, strengthening the local economy, and decreasing environmental pollution.