The potential for nurturing, integrating, and optimizing physical literacy to thereby enhance the quality of life, but supporting such arguments remains limited. The purpose of this study was to investigate the mediation effect of motivation in the relationship between physical literacy and quality of life. By applying the time segregation method, participants (n = 388) were drawn from university students of Taiwan who were asked to complete a set of questionnaires twice in the first semester of the academic year 2021–2022. Partial least square structural equation modelling was used to test the hypotheses. Results further indicated that motivation is the mediator of the relationship between physical literacy and physical education satisfaction (direct effect = 0.37, p < 0.001). This satisfaction also mediated the relationship between physical literacy and quality of life (direct effect = 0.592, p < 0.001). This study connected theoretical knowledge regarding physical literacy with practice, suggesting that ongoing physical education may nurture the habit of lifelong participation in physical activity, and identifies the critical role of motivation as a mediating mechanism, thus further improving quality of life.
