In the present study, antioxidant and antimicrobial potential of the Lentinus tigrinus (Bull.) Fr. mushroom was determined. Total antioxidant status (TAS), total oxidant status (TOS), and oxidative stress index (OSI) of the mushroom were measured with Rel Assay kits. Antimicrobial activities were tested on 9 standard bacterial and fungal strains (Staphylococcus aureus, Staphylococcus aureus MRSA, Enterococcus faecalis, Escherichia coli, Pseudomonas aeruginosa, Acinetobacter baumannii, Candida albicans, Candida krusei, and Candida glabrata) with a modified agar dilution method. It was determined that the TAS value of L. tigrinus was 1.748 ± 0.071, TOS value was 19.294 ± 0.237, and OSI was 1.106 ± 0.031. It was also found that mushroom extracts generally exhibited higher activity on Candida albicans, C. krusei, and C. glabrata. In conclusion, it was suggested that L. tigrinus can be used as a natural source due to its antioxidant and antimicrobial activities.