ABSTRACT. Submerged cultivation of medicinal basidiomycetes is a reproducible and efficient method of producing mycelia and metabolites. The antioxidant activity indicates its medicinal properties and is an important tool for basidiomycete screening. In this study, we analyzed the production of mycelial biomass and exopolysaccharides and the antioxidant activity of basidiomycete strains in submerged cultivation. Twenty-five strains were used for submerged cultivation in extract malt medium, and the production of mycelial biomass and exopolysaccharides was evaluated. Antioxidant activity was determined using the 1,1-diphenyl-2-picrylhydrazyl method. Among the 25 evaluated strains, Lentinus crinitus produced the highest biomass, reaching 1190 ± 52 mg·L -1 day -1 ; Agaricus subrufescens strains had the highest exopolysaccharide production from 18.96 ± 0.15 to 20.97 ± 2.10 mg L -1 ·day -1 . Additionally, A. subrufescens showed the highest total antioxidant activity, reinforcing the therapeutic potential of this basidiomycete. No significant correlation was found between mycelial biomass or exopolysaccharide production and antioxidant activity; S.H. Umeo et al. 9908©FUNPEC-RP www.funpecrp.com.br Genetics and Molecular Research 14 (3): 9907-9914 (2015) however, the results depended on each species and the strains of the same species. We found large variations in the production of mycelial biomass and exopolysaccharides and in antioxidant activity among different species and among strains of the same species. Thus, evaluating the total antioxidant activity is an important tool for identifying strains with biotechnological potential.