The objective of this study is to investigate the impact of servant leadership on employee well-being in the Jordanian hospitality sector. Furthermore, it attempted to explore the potential mediation effects of job burnout, role overload, and anxiety on this relationship. The data in this study were obtained from a sample of 341 individuals who were employed in five-star hotels located in Amman using a simple random sample. For analysis, the study employed the SEM-AMOS software. The results indicated a noteworthy and favorable influence of servant leadership on the well-being of employees (p = 0.001), emphasizing its crucial meaning in creating a nurturing work atmosphere that promotes enhanced well-being among employees. Additionally, the study has identified anxiety and job burnout as a mediator (p = 0.001), indicating that servant leadership has an impact on well-being by reducing levels of anxiety and job burnout. Nevertheless, the hypothesized mediating effect of role overload was not supported (p = 0.070) in the specific setting under investigation. The study’s theoretical implications contribute to the broader comprehension of the complex connections between servant leadership, employee well-being, and mediating variables. Furthermore, it provides empirical insights within the specific context of the Jordanian hospitality business.