The objectives of this research work were to evaluate the effect of in vitrogastrointestinal digestion (GIT) on melon peel juice (MPJ) powder from fruit processing industry by-products, considering (i) the recovery and accessibility indexes, (ii) the changes on antioxidant activity, and (iii) the prebiotic effect. Throughout exposition to GIT conditions a decrease on the total phenolic content (TPC = 65.31%) and antioxidant activity by ABTS = 39.77% and DPPH = 45.91% were observed. However, these both parameters exhibited stable accessibility, accounting with 81.89%, 76.55%, and 54.07% for TPC, ABTS and DPPH, respectively. After gastrointestinal digestion, the non-absorbed fraction exhibited a positive impact on the growth of Bifidobacteriumand Lactobacillusstrains, possibly associated with the high content of simple sugar (glucose and fructose). This fraction also showed to be safe on Caco-2 intestinal cells. These findings suggest that MPJ might be used as a potential food functional ingredient.