Background <p>Most nursing assistants in Macao are migrant workers from mainland China. They face tremendous job stress as engaging in long-term, repetitive, and lower-end caring work. However, empirical studies examining the stress–burnout pathway in the Chinese settings remain scarce. This study aims to investigate the relationship between job stressors and burnout among nursing assistants in Macao, China, and further explores the moderating role of organizational culture in the association.</p> Method <p>This study adopted the quantitative research method, and 290 samples were collected from nursing assistants in six local residential care homes for older people. Among the participants, 78.6% were female, and 72.8% were from Mainland China.</p> Results <p>Using structural equation modeling, this study found that perceived job stressors were significantly associated with job burnout. The finding revealed the serial mediation of emotional exhaustion and depersonalization in the pathway from perceived job stressors to personal accomplishment. The study also showed that organizational culture moderated the relationship between perceived job stressors and emotional exhaustion. The association between job stress and emotional exhaustion is weakened when organizational culture is high, while the association is strengthened when organizational culture is low.</p> Conclusions <p>The study findings underscore the need for early intervention and optimized management practices to prevent severe job burnout among nursing assistants. Moreover, it is essential for organization leaders to recognize job stressors and prioritize addressing emotional strain and depersonalization issues among nursing assistants and develop a positive organizational culture to help them effectively engage in geriatric care work.</p>