Background: Musculoskeletal pain is common in the working population and may affect the work ability, especially among those with high physical work demands. This study investigated the association between physical work demands and work ability in workers with musculoskeletal pain. Methods: Workers with physically demanding jobs (n = 5377) participated in the Danish Work Environment Cohort Study in 2010. Associations between physical work ability and various physical work demands were modeled using cumulative logistic regression analyses while controlling for possible confounders. Results: In the fully adjusted model, bending and twisting/turning of the back more than a quarter of the workday (reference: less than a quarter of the workday) was associated with higher odds of lower work ability in workers with low-back pain (OR: 1.38, 95% CI: 1.09-1.74) and neck-shoulder pain (OR: 1.29, 95% CI: 1.01-1.64). When adding up the different types of demands, being exposed to two or more physical work demands for more than a quarter of the workday was consistently associated with lower work ability. Conclusions: Work that involves high demands of the lower back seems especially problematic in relation to work ability among physical workers with musculoskeletal pain. Regardless of the specific type of physical work demand, being exposed to multiple physical work demands for more than a quarter of the workday was also associated with lower work ability.