Hypsizygus tessulatus also known as buna-shimeji is an endemic Japanese edible and medicinal mushroom. Attempt to grow this species under Philippine condition was undertaken from November 2013-March 2014 and grown under laboratory condition using different cultural parameters and its antibacterial activity was also determined. Different culture media, temperature and pH, spawning and growing substrates are utilized. The highest rate of radial mycelia growth among ten different culture media was observed in compost agar measuring (78.67 mm) in diameter, optimum level of pH (6 and 7) and temperature (20-25° C) were attained. Profuse mycelia colonization was attained on 18 days after inoculation using black beans as spawning substrate however, sorghum spawn inoculated in paddy straw and sawdust- rice bran was found to be the most suitable fruiting substrate for initiating mycelia growth (135 mm). Methanol extract of oven-dried mycelia mat of H. tessulatus (HMME) was used against two Gram positive (+) and Gram negative (-) bacteria for antibacterial study. An average inhibition zone exhibited by H. tessulatus mycelia extract in Staphylococcus aureus (15.67 mm). H. tessulatus exemplifies moderate and fair antibacterial activities.Vegetative studies show that H. tessulatus grows at optimum temperature of 25-27° C under laboratory condition. Furthermore, buna-shimeji mushroom cultivation requires for its fructification stage in lower temperature and higher relative humidity.