Recently, adaptive non-shivering thermogenesis has attracted considerable attention because it can elevate energy expenditure and help treat obesity. Despite the numerous reports related to UCP1-driven thermogenesis, little is known regarding UCP1-independent thermogenesis in adipose tissues and muscle. Therefore, it is essential to identify the molecular targets for UCP1-independent thermogenesis and their mechanisms to increase the energy expenditure pharmacologically in both adipocytes and muscle. This study examined whether trans-anethole (TA), a major component of the essential oils of fennel, induces UCP1-independent SERCA/SLN-based thermogenesis and promotes the lipid metabolism in muscle cells. TA enhanced myogenesis, lipolysis, and the oxidative metabolism in C2C12 muscle cells. More importantly, TA activated the SERCA/SLN/RYR axis, thereby inducing thermogenesis in muscle cells. Molecular docking analysis revealed a good interaction between SERCA with TA with a strong bind activity. In conclusion, the current data unveiled a previously unknown mechanism of TA in myoblasts and suggests a possible therapeutic agent in muscles by enhancing energy expenditure.