An appropriate level of physical activity (PA) is beneficial for health and weight management. This study examines the relationship of habitual PA and activity energy expenditure (AEE) in a typical week with gender and body composition parameters. Data of habitual PA of 72 healthy adults were collected with the heart rate and motion sensor (Actiheart). Total and segmental body fat was measured with bioelectrical impedance analyser. Men were more active in total and on working days and had shorter light and longer moderate and vigorous PA on working days than females (p<0.05). Females had less inactive and sitting time on days off. Inactive and sitting time in working days for both genders correlated positively with visceral fat rating, body and leg fat % (p<0.05). In male’s duration of working days’ moderate PA correlated inversely, but light PA positively with these parameters (p<0.05). Males with more vigorous PA in working day and AEE in days off had lower body fat % (p<0.05). A distinctive PA pattern on working days and days off indicates a gender-related differences. Decrease of time of inactivity and light PA and increase of duration of moderate and vigorous PA was associated with lower body and segmental fat amount.