Macrolepiota procera, commonly called the Parasol Mushroom, is a delicious mushroom collected from the nature and commonly consumed by the public in many regions of Turkey. This study was conducted to determine the optimum culture conditions (pH, temperature, carbon and nitrogen sources) for mycelial growth of M. procera. Three pH values (pH 5.0, 5.5 and 6.0), four incubation temperatures (15, 20, 25 and 30°C), seven carbon (C) sources (dextrose, glucose, lactose, maltose, mannitol, sucrose and xylose) and six nitrogen (N) sources ((NH₄)₂HPO₄, NH₄NO₃ and Ca(NO₃)₂, malt extract, peptone and yeast extract) were investigated. In the second step of the study, the effect of seven pH values (4.0, 4.5, 5.0, 5.5, 6.0, 6.5 and 7.0) on the mycelial colony diameter was examined at 20 and 25°C since these temperatures gave the best mycelial growth in the previously conducted temperature experiment. The best mycelial growth was determined at pH 6.0. The optimum temperature for mycelial growth of M. procera was found as 25°C. The use of glucose as carbon source and yeast extract and peptone as nitrogen source in the culture medium gave the best results for mycelial growth. Determining of optimum culture conditions for mycelial growth of M. procera will provide important contributions to the fortcoming studies on it’s commercially cultivation in Turkey.