Antibacterial activity of N^α-lauroyl-L-arginate ethylester(LAE) against five pathogenic microorganisms of fruits and vegetables were investigated in this study. Bacteriostatic test in vitro was taken to research the antibacterial activity of LAE. Penicillium digitatum, Penicillium italicum, Botrytis cinerea, Alternaria alternate and Pectobacterium carotovorum subsp. Carotovorum were treated as the model bacteria of pathogenic microorganisms of fruits and vegetables, and the minimum inhibitory concentration(MIC) of LAE against these model bacteria were determined. The inhibition of LAE to the hypha growth for moulds and the bactericidal effect of LAE for bacteria were also determined. Then the combined antibacterial effects of LAE and other preservative were explored. The results showed that the MIC of LAE against Botrytis cinerea, Alternaria alternate, Penicillium italicum, and Penicillium digitatum were 400, 200, 400, 400 μg/mL respectively. When the concentration of LAE reached the MIC of each strain, the high mycelial inhibition rate of the four molds could be maintained at 100%（16 h）, 78.2%(48 h), 81.8%(48 h), and 79.1%(48 h) respectively. The MIC of LAE against Pectobacterium carotovorum subsp. Carotovorum was 25 μg/mL, and LAE with the concentration of 50 μg/mL could completely kill the tested bacteria within 90 min. Morever, the combination of LAE and sodium chloride could exert the best inhibitory effect on the model bacteria.