Foods are vulnerable to contamination by pathogenic bacteria during production, transportation and storage, and this can cause foodborne illness. Linalool has antibacterial effects, but the mechanism of action remains unclear. In order to provide a theoretical basis for the development of linalool as a natural food preservative, the antibacterial activity and mechanism of linalool against Pseudomonas fragi were studied. The antibacterial activity of linalool was evaluated by measuring the minimum inhibitory concentration and plotting the bacterial growth curve. The antibacterial mechanism was investigated by scanning electron microscope observation, crystalline violet staining experiments, fluorescein diacetate staining experiments, and measuring the changes in electrical conductivity, nucleic acid leakage, respiratory metabolic activity and respiratory chain dehydrogenase activity. The results showed that linalool had a strong inhibitory effect against Pseudomonas fragi, and the minimum inhibitory concentration was 1.5mL/L. Linalool could damage the structure morphology and cell membrane of Pseudomonas fragi, and improve the permeability of the cell membrane, thus leading to the leakage of intracellular materials and the increase of electrical conductivity outside the cell membrane. It could inhibit the activity of respiratory metabolic and respiratory chain dehydrogenase, and damage the respiratory chain, thus leading to the disorder of intracellular metabolism. The research confirmed that linalool exerted a antibacterial effect by damaging the cell structure of Pseudomonas fragi and inhibiting its respiratory metabolism, and linalool could be studied and developed as a natural preservative.