Resumen: Boletus edulis is a widely consumed edible mushroom, known for its high nutritional value and bioactive compounds of interest. This study aimed to explore its potential bioactive properties by evaluating the inhibitory effects of B. edulis extract on key digestive enzymes and its antioxidant capacity through in vitro assays. For this purpose, an hydroethanolic extract of B. edulis was evaluated phytochemically and through in vitro bioassays for antioxidant activity and enzyme inhibition. The extract contained 226 ± 3 mg of ergosterol/100 g, and phenolic compounds such as epigallocatechin-3-gallate (501 ± 18 mg/100 g) and caffeoyl tryptophan (15 ± 1 mg/100 g), consistent with fungal species of functional interest. Cytotoxicity effects of the extract were assessed in HepG2 cells, and results showed no significant effect on the loss of cell viability. Furthermore, it showed antioxidant capacity, including superoxide radical scavenging, nitric oxide and intracellular ROS production as well as a moderate inhibition of metabolic enzymes such as α-glucosidase (IC₅₀ = 13 ± 1 mg/mL) and lipase (IC₅₀ = 25 ± 11 mg/mL), and the pro-inflammatory enzyme 5-lipoxygenase, with an approximate IC₅₀ value of 2.5 mg/mL. These results evidence a multi-targeted bioactive profile that could offer cumulative health benefits with regular consumption. However, further in vivo and clinical studies are necessary to confirm its potential as functional food. Idioma: Inglés DOI: 10.1016/j.afres.2025.101342 Año: 2025 Publicado en: Applied food research 5, 2 (2025), 101342 [10 pp.] ISSN: 2772-5022 Financiación: info:eu-repo/grantAgreement/ES/DGA/B44-23R Tipo y forma: Article (Published version) Área (Departamento): Área Fisiología (Dpto. Farmac.Fisiol.y Med.L.F.)