In order to develop the natural food preservative, freeze dried plantain (Plantago asuaica L.) was extracted with several solvents, and antimicrobial activity was investigated. The methanol extract from the plantain exhibited antimicrobial properties against five strains such as Bacillus subtilis. Escherichia coli, Staphylococcus aurcus, Listeria monocytogenes, and Vibrio parahaemolyticus. The methanol extract at the concentration of 1.0㎎/㎖ completely inhibited the growth of B. subtilis and V. parahaemolyticus. Antimicrobial activity of the ethylacctate fraction from the methanol extract of plantain was the strongest fraction compare to those of the other solvent fractions such as n-hexane, chloroform, n-butanol and water. The ethylacetate fraction showed the inhibitory effect at the concentration of 0.5㎎/disc on the growth of the B. subtilis and V. parahaemolyticus.