Surface electromyography (EMG) has been used to estimate muscle work and physiological burden of the whole body during human movements. However, there are spatial variations in surface EMG responses within individual muscles. The aim of this study was to investigate the relation between oxygen consumption and surface EMG responses of lower leg muscles during walking at various speeds and to quantify its spatial variation within an individual muscle. Nine young males walked on a treadmill at four speeds: preferred minus 1 km/h, preferred, preferred plus 1 km/h, and preferred plus 2 km/h, and the metabolic response was measured based on the expired gas. High-density surface EMG of the tibialis anterior (TA), medial gastrocnemius (MG), lateral gastrocnemius, and soleus muscles was performed using 64 two-dimensional electrode grids. Correlation coefficients between oxygen consumption and the surface EMG amplitude were calculated across the gait speeds for each channel in the electrode grid and for individual muscles. Mean correlation coefficients across electrodes were 0.69–0.87 for the four individual muscles, and the spatial variation of correlation between the surface EMG amplitude and oxygen consumption within an electrode grid was significantly greater in MG muscle than in TA muscle (Quartile deviations: 0.24 for MG and 0.02 for TA, p < 0.05). These results suggest that the physiological burden of the whole body during gait at various speeds can be estimated from the surface EMG amplitude of calf muscles, but we need to note its spatial distribution within the MG muscle.