Ethyl acetate, methanol, water extract and their fractions from chestnut flower(Castancea crenata) were tested for antimicrobial activities. Yields of prebloomed chestnut flower extracts were 13.84, 12.90 and 1.82% in methanol, water and ethyl acetate, and those of the postbloomed were 13.12, 11.75 and 1.18%, respectively. Methanol extract from the chestnut flower was fractionated by solvents using hexane, chloroform, ethyl acetate, n-butanol and water, yields of those fractions were 0.16, 0.08, 1.94, 4.75 and 6.91% in the prebloomed, and were 0.90, 0.13, 1.40, 3.42 and 7.18% in the postbloomed. In the solvent extracts of water, ethyl acetate and methanol, methanol extract showed the most effective antimicrobial activity, antimicrobial activity of ethyl acetate fraction of methanol extract was stronger than others. Minimum inhibitory concentration of ethyl acetate fractions from the prebloomed showed 100, 140, 100 and 90ppm against Staphylococcus aureus, Streptococcus mutans, Echerichia coli and Salmonella typhimurium, in fractions from the postbloomed were 140, 140, 100 and 150ppm, respectively. Growth of all the strains was completely inhibited to 30 hours in a 150ppm concentration. E. coli sells treated with ethyl acetate fraction was collapsed severely.