In this study, the potential of Moso bamboo sawdust as an alternative substrate for the cultivation of oyster mushroom (Pleurotus ostreatus) was investigated. Oyster mushroom was cultivated on 2-months fermented bamboo sawdust (BS) and mixed with rice bran (RB) and sweet potato schochu lees (SPSL) as additional nutrition. The growth condition, morphological properties, nutritional, mineral contents and free amino acid content of mushroom cultivated were evaluated. Based on the results, the total growth days on the bamboo media were between 3 and 7 days shorter than the conventional media. The bamboo media mixed with RB had better yield and fruiting bodies at 97.9 ± 3.9 g/bottle and 33.6 ± 4.2 no/bottle, respectively. Furthermore, the addition of SPSL to BS increased the protein content and decreased the carbohydrate contents of fruit bodies. In addition, the free-amino acids in the fruit bodies from the bamboo media were 1.5 times higher than the conventional media, which potentially added the higher value to usual mushrooms. Hence, oyster mushroom cultivation can be an alternative method to reduce bamboos wastes in Japan and would promote sustainable growth in agricultural industry.