Older females experience higher rates of disability than males, potentially due to sex-specific differences in gait and muscle function. The authors evaluated the effects of age and physical activity (PA) on gait mechanics and knee extensor muscle function in males and females. Three groups of 20 individuals (each 10 females) participated: young (21–35 y) and highly and less active older (55–70 y) adults. Knee extensor strength and joint mechanics during preferred speed gait were collected before and after 30 minutes of walking. Age by sex and PA by sex interactions indicated older and less active older females had lower concentric knee extensor muscle power and larger hip extension moments than males. After 30 minutes of walking, older less active adults had larger decreases in knee extensor power than their highly active older counterparts, and older adults of both sexes had decreases in ankle dorsiflexion moments while young adults did not. These results suggest that older, particularly less active, adults are susceptible to knee extensor muscle fatigue from moderate activity. For older adults, high levels of PA may be necessary to preserve gait mechanics in response to a bout of exercise. This new information may be important for targeting interventions in at-risk older adults.