This study was carried out to develop artificial culture method of Grifola frondosa in polypropylene bag. To find out the favorable substrate formulation of G. frondosa, physicochemical conditions were investigated. The T2 formulation (55 : 25 : 12 : 8) mixing ratio of oak sawdust, oak chip, dried bean-curd refuse and wheat bran showed the shortest time to complete the crop cycle and the highest yield (weight of fresh mushrooms harvested at maturity). Those physicochemical conditions were pH 4.3, 2.4% crude fat contents, 54.1% total carbon, 1.42% total nitrogen, 38.1 C/N ratio, 75.5% porosity and 0.21 g/cm ³ bulk density. Among the physicochemical factors, pH, crude fat and total nitrogen may affect mushroom yield. Therefore, development of favorable substrate would be benefit to increase production efficiency of G. frondosa mushrooms and be commercial potential.