Background:The pandemic has drastically impacted lifestyles, including the frequency, duration, and intensity of physical activity, which, in turn, leads to negative effects on job burnout among healthcare workers. Objective: This study evaluated the association between the level of physical activity and job burnout in a sample of health professionals during health emergency. Methods: In this cross-sectional study, 300 workers working in a public hospital, located in the city of Rioja, Department of San Martin, Peru, were considered. Data were collected through an online and face-to-face survey. Job burnout was assessed using a validated questionnaire adapted to the Peruvian population and physical activity was assessed based on a single item. Crude and adjusted linear regression models were created to assess the association between the frequency of physical activity performed weekly and the level of job burnout. Results: In the crude models, it was observed that participants who performed physical activity 1-2 times, 3-4 times, or more than 4 times per week had, respectively, 6.29 (95% CI, 11.00-1.59), 14.28 (95% CI, 21.70-6.85), and 14.95 (95% CI, 25.17-4.72) lower job burnout points compared to those who never performed physical activity. In the model adjusted for confounding variables, the association of this result was maintained, showing a dose-response effect. Conclusion: These findings suggest the need for the implementation of health education programs that include physical activity to minimize the negative effects of job burnout and ensure better medical care by health professionals for the benefit of patients.