This work aimed to investigate the inactivation mechanism of lauroyl arginate ethyl (LAE) against Listeria monocytogenes. The antibacterial activity of LAE against L. monocytogenes was evaluated by measuring the minimum antibacterial concentration (MIC). Then, the antibacterial mechanism of LAE against L. monocytogenes was investigated by measuring the cell morphology, cell membrane integrity, intracellular ATP level, cell membrane potential, cell surface hydrophobicity, and intracellular reactive oxygen species (ROS) level. The results showed that LAE could effectively inhibit the growth of L. monocytogenes with a MIC value of 10 μg/mL. After LAE treatment at a final concentration of 40 μg/mL for 10 min, the cell morphology of L. monocytogenes shrank obviously, the levels of extracellular nucleic acid and protein significantly (P