The development of natural antimicrobial agents offers new strategies for food preservation due to the health hazards associated with the spoilage of meat products caused by microbial contamination. In this paper, the inhibitory mechanism of protocatechualdehyde (PCA) onListeria monocytogeneswas described, and its effect on the preservation of cooked chicken breast was evaluated. The results showed that the minimal inhibitory concentration (MIC) of PCA onL. monocytogeneswas 0.625 mg/mL. Secondly, PCA destroyed the integrity of theL. monocytogenescell membrane, which was manifested as a decrease in membrane hyperpolarization, intracellular ATP level, and intracellular pH value. Field emission gun scanning electron microscopy (FEG-SEM) observed a cell membrane rupture. Transcriptome analysis showed that PCA may inhibit cell growth by affecting amino acid, nucleotide metabolism, energy metabolism, and the cell membrane ofL. monocytogenes. Additionally, it was discovered that PCA enhanced the color and texture of cooked chicken breast meat while decreasing the level of thiobarbituric acid active substance (TBARS). In conclusion, PCA as a natural antibacterial agent has a certain reference value in extending the shelf life of cooked chicken breast.