OBJECTIVE The aim of the present study is to examine the association between physical activity and perceived job demand, job control, perceived stress and energy, and physiological arousal reflected by morning and evening concentrations of cortisol in saliva among white-collar workers. METHODS Physical activity during the last week was assessed during work and leisure time by a Danish version of the International Physical Activity Questionnaire and saliva samples were collected. The study group comprised 389 white-collar workers, aged 25-67 years, and of which 257 were women. RESULTS We found that physical activity during leisure time was associated with higher perceived energy, and for men also with lower perceived stress. Further, we found that physical activity at leisure time affected the association between salivary cortisol and perceived stress and energy so that respondents being physically active at leisure time and perceiving higher energy showed higher evening saliva cortisol. CONCLUSIONS Physically active employees perceive less stress and more energy. The association between stress-energy and salivary cortisol was affected by vigorous physical activity. No association between job control-demand and the degree of physical activity was found. Based on the present data, we recommend office workers-exposed to high job strain and inactivity at the job-to perform physical activity, preferably of high intensity, in order to reduce stress and increase energy.