Muscle synergy analyses are used to enhance our understanding of motor control. Spatially fixed synergy weights coordinate multiple co-active muscles through activation commands, known as activation coefficients. To gain a more comprehensive understanding of motor learning, it is essential to understand how activation coefficients vary during a learning task and at different levels of movement proficiency. Participants walked on a line, a beam, and learned to walk on a tightrope—tasks that represent different levels of proficiency. Muscle synergies were extracted from electromyography signals across all conditions and the number of synergies was determined by the knee-point of the total variance accounted for (tVAF) curve. The results indicated that the tVAF of one synergy decreased with task proficiency, with the tightrope task resulting in the highest tVAF compared to the line and beam tasks. Furthermore, with increasing proficiency and after a learning process, trial-to-trial similarity increased and temporal overlap of synergy activation coefficients decreased. Consequently, we propose that precise adjustment and refinement of synergy activation coefficients play a pivotal role in motor learning.