Objective: To determine whether Lentinula edodes (L edodes) cultured on eucalyptus chips in the Ecuadorian highland generates antioxidant metabolites and anti-inflammatory effects. Methods: Total phenolic content was determined by the Folin-Ciocalteu reaction, the in vitro antioxidant activity of L edodes extracts was evaluated by the DPPH method and the in vivo anti-inflammatory activity was studied in mice. The chemical composition was studied by phytochemical screening and gas chromatography/mass spectrometry (GC/MS). Results:Total phenols were higher in extracts with increasingly high-water content. The antioxidant activity was robust and significantly strong in these extracts, suggesting the active metabolites are water-soluble. The anti-inflammation activity was significant in aqueous extracts only. Phytochemical screening indicated an overall similar composition to the literature reported earlier. GC/MS detected galactitol, trehalose, xylitol, phosphoric acid and octadecanoic acid among the most abundant metabolites. Conclusions: Cultivation on eucalyptus biomass at the Ecuadorian highlands retains the overall chemical composition, the phenolic content, antioxidant levels, and the in vivo anti-inflammatory activity of L edodes. The 11.3% content of trehalose observed is interesting for its capacity to control cellular stress damage. [ABSTRACT FROM AUTHOR]