Building on the conservation of resources and self-determination theories, this study investigated the psychological mechanisms of how job autonomy and stress jointly influence the well-being of hotel employees via two psychological paths. Using a multi-stage research design with data collected in three waves from 627 frontline employees working in hotels in China, the findings reveal several key results. First, job stress negatively impacts employee well-being through the mediating role of surface acting, whereas job autonomy positively impacts well-being through the mediating role of deep acting. Second, the interaction between job stress and job autonomy significantly buffers the negative impact of stress on well-being. Third, prosocial motivation serves as a moderator, weakening the relationship between job stress and surface acting while strengthening the relationship between job autonomy and deep acting. Furthermore, prosocial motivation plays a moderated mediation role in both pathways. The study enriches the literature on job autonomy and well-being and contains important theoretical and managerial implications.