Ipomoea turbinata Lagasca and Segura (Purple Moonflower) belongs to the largest flowering genus Ipomoea in the Convolvulaceae family. Ipomoea turbinata has not been previously explored for its in vitro potential. This is the first study focused on thidiazuron-induced callus culture for efficient biosynthesis of commercially important phenolic compounds in this plant species. Among the two plant growth regulators tested on leaf, stem, and root explants, 5 mg L−1 thidiazuron (TDZ) induced the highest biomass accumulation (61.4 g L−1 fresh weight, 6.3 g L−1 dry weight) in leaf-derived callus cultures after 5 wk of culture. The highest total phenolic and flavonoid contents recorded were 9.04 mg g−1 and 1.16 mg g−1, respectively, in optimized callus cultures. High-performance liquid chromatography analysis indicated high levels of pharmacologically important anticancer compounds such as chlorogenic acid (13.48 mg g−1), arctigenin (11.67 mg g−1), quercetin (6.19 mg g−1), and kaempferol (5.48 mg g−1), along with other phenolic acids. Furthermore, the antioxidant activity was also evaluated, and leaf-derived callus culture displayed a maximum of 62.6% antioxidant potential. The induction of improved biomass accumulation in callus culture and the production of multipotent bioactive metabolites shows the potential of the multifunctional thidiazuron hormone as an efficient elicitation tool in callus culture of I. turbinata.