Objectives The aim of this study was to evaluate the effects of a portable pedal machine intervention (60 minutes per working day) for 12 weeks on healthy tertiary employees' cardiometabolic risk factors. Methods Anthropometric parameters, body composition, cardiometabolic/inflammatory markers, physical fitness, physical activity, and sedentary time measured before and after the intervention were compared between office healthy workers who used a portable pedal machine (INT, n = 17) and those who did not (CTRL, n = 15). Results The INT group improved Delta ultrasensitive C-reactive protein (P = 0.008), Delta total cholesterol (P = 0.028), and Delta light-density lipoprotein cholesterol (P = 0.048) compared with the CTRL group (Delta: T1-T0). The intervention reduced daily sitting time (P <= 0.01) and increased time spent at light intensity (P <= 0.01) and moderate-to-vigorous (P <= 0.01) physical activity compared with baseline values. Conclusions These findings suggest that promoting physical activity during workdays can reduce the negative health effects of spending too much time sitting and inactive.