This study aimed to determine physical performance criteria of different occupational groups by investigating physical activity and energy expenditure in healthy Swiss employees in real-life workplaces on workdays and non-working days in relation to their aerobic capacity (VO2max). In this cross-sectional study, 337 healthy and full-time employed adults were recruited. Participants were classified (nine categories) according to the International Standard Classification of Occupations 1988 and merged into three groups with low-, moderate- and high-intensity occupational activity. Daily steps, energy expenditure, metabolic equivalents and activity at different intensities were measured using the SenseWear Mini armband on seven consecutive days (23 hours/day). VO2max was determined by the 20-meter shuttle run test. Data of 303 subjects were considered for analysis (63% male, mean age: 33 yrs, SD 12), 101 from the low-, 102 from the moderate- and 100 from the high-intensity group. At work, the high-intensity group showed higher energy expenditure, metabolic equivalents, steps and activity at all intensities than the other groups (p<0.001). There were no significant differences in physical activity between the occupational groups on non-working days. VO2max did not differ across groups when stratified for gender. The upper workload limit was 21%, 29% and 44% of VO2max in the low-, moderate- and high-intensity group, respectively. Men had a lower limit than women due to their higher VO2max (26% vs. 37%), when all groups were combined. While this study did confirm that the average workload limit is one third of VO2max, it showed that the average is misrepresenting the actual physical work demands of specific occupational groups, and that it does not account for gender-related differences in relative workload. Therefore, clinical practice needs to consider these differences with regard to a safe return to work, particularly for the high-intensity group.