Objective: To examine the effects of different shoe weights on lower leg muscle fatigue when walking by electromyographic(EMG) analysis due to the most effective weight for loading not being established. Design: Cross-sectional study. Methods: Thirty healthy university students (15 male, 15 female) were enrolled and randomly assigned into three conditions, which included wearing athletic shoes with an addition of 300 g, 500 g, and 1,000 g weights respectively. Prior to walking, all subjects were instructed to sit in a chair for 10 minutes. All subjects walked at a speed of 3.6 m/s on a treadmill for 20 minutes without rest. EMG measurements were taken using the median power frequency to assess for the effect of the different weight of shoes on muscle fatigue of the soleus, gastrocnemius, and tibialis anterior while walking on a treadmill in an upright posture. EMG measurements were taken during the first and last 30 seconds of walking. Results: In terms of muscle fatigue, for the soleus, the median power frequency was significantly lower with 1,000 g compared with 300 g and 500 g (p<0.05). For the tibialis anterior, the median power frequency was significantly lower with 1,000 g than 300g and 500 g (p<0.05). For the gastrocnemius, the median power frequency was significantly lower with 1,000 g compared with 300g (p<0.05). Conclusions: Increased shoe weight increases soleus, gastrocnemius, and tibialis anterior muscle fatigue during ambulation.