Lentinus tigrinus is a common wood-decaying basidiomycete that has a fleshy pileus, strong odor and agreeable taste. In this study, the effect of various sawdust-based substrates from different wood species (poplar, oak, hornbeam and ash) and supplementary wheat bran at the various levels (0, 1, 5 and 10%) on spawn run times, yield, fruiting body size and protein content of Lentinus tigrinus were investigated. The strain of L. tigrinus, originally isolated from the Haciosman forest in Samsun, was used. Some chemical properties of the substrates such as pH, moisture, nitrogen and carbon content, carbon: nitrogen ratio, and spawn run time, mushroom yield and properties of fruiting body were determined. Dry matter and protein contents of fruiting bodies were also established. Among wood species, the longest spawn run time was found in the poplar sawdust-based substrate. Spawn run time shortened and mushroom yields increased with an increase in the rate of wheat bran in the mixtures. Oak, hornbeam and ash sawdust-based substrates and supplementary wheat bran at the rate of 5 and 10% were found to be suitable for favorable mycelium growth and high mushroom yield. The effect of various sawdust-based substrates from different wood species on fruiting body sizes was found to be significant.