In this study, the posibility of using of waste lime leaves in Pleurotus ostretatus cultivation was investigated. After a successful harvest, total phenolic content, antioxidant and antimicrobial properties of mushrooms was determined. According to the test results; yield was 15%, biological efficiency was 30%. The total phenolic content was 151.4±0.001 mg GAE/100 g, the antioxidant activity was 2,508±0.056 μmol FeSO4.7H2O/g. Additionally; methanolic extract of Pleurotus ostreatus mushroom showed inhibitory effect against Klebsiella pneu- monia and Acinetobacter haemolyticus bacteria. As a result, Tilia Tomentesa’s leaves are suitable raw material with satisfactory medical properties for the Pleurotus ostretaus cultivation.