AbstractThis study investigated the impact of pulsed light (PL) applied at a fluence of 4–12 J cm−2 (complying with U.S. Food and Drug Administration standards) on fresh‐cut watermelon, focusing on its efficacy in inactivating various food‐borne pathogenic gram‐positive (Escherichia coli and Staphylococcus aureus) and gram‐negative (Listeria monocytogenes and Salmonella) bacteria. The inactivation data were used to fit the log‐linear and Weibull kinetic models. Additionally, the impact of PL on physicochemical properties, lycopene content, total phenolic content, antioxidant activity, and pectin methylesterase (PME) residual activity (RA) was studied. The results demonstrated that the inactivation rate increases with the increase in fluence. Salmonella was most sensitive to PL, resulting in a reduction of 2.55 ± 0.08 log CFU g−1 and a higher kmax, whereas S. aureus was most resistant, with a reduction of 1.87 ± 0.24 log CFU g−1 and a lower kmax. The Weibull model was the best to predict the inactivation kinetics based on standard statistics and information theory criteria. For all assayed food‐borne pathogens, the inactivation curve showed non‐linearity with upward concavity except for L. monocytogenes, which followed a concave downward curve (β >1). Results showed that PL was effective in the retention of color, firmness, pH levels, total soluble solids, and titratable acidity, and also had a positive effect on enhancing the levels of essential components such as lycopene, total phenolic compounds, and antioxidant activity. Additionally, the PME RA significantly decreased with an increase in fluence due to structural modification. Therefore, PL can be explored to enhance fresh‐cut watermelon's safety and quality attributes.Practical ApplicationsThe implementation of this research holds significant potential for practical benefits in the fresh‐cut fruit industry. The findings could revolutionize processing practices, ensuring enhanced microbial safety, enzyme inactivation, and an extended shelf‐life of fresh‐cut watermelon. The study strongly contributes to preserving the desirable texture, color, and overall quality of the product, enhancing the retention of lycopene and other valuable antioxidants by standardizing the pulsed light treatment parameters while adhering to U.S. Food and Drug Administration standards for food application. Moreover, this research will scientifically contribute to addressing the critical challenges of foodborne outbreaks in the fresh‐cut fruit sector while providing consumers with a healthy, convenient food choice.