Walking is a complex motor skill involving significant interaction within the CNS and various biomechanical elements. Asymmetry is one of the most apparent characteristics of walking, which is controlled by muscle synergies. The asymmetry in muscle synergies during walking is not well understood. This study aimed to investigate limb asymmetry in synergistic control of the selected lower limb muscles during walking. The study included sixteen healthy male participants, and bilateral lower limb muscle activities were recorded during walking. Muscle synergies and relative muscle weights were extracted using non-negative matrix factorization. The results showed that for both dominant and non-dominant legs five synergies are sufficient to reconstruct the original signal. Activation patterns in the first, fourth, and fifth synergies showed high similarity, with differences in relative muscle weights observed in the first and third synergies. However, in the second synergy, the similarity of activation patterns between the legs was small with variations in specific lower limb muscles. These findings suggest that synergy structure is independent of leg dominance, but the similarities of activation patterns range from small to high between dominant and non-dominant legs.