This study investigated the impact of chemical inducers on the co-production of yeast oil (YO) and β-carotene from purified biodiesel-derived crude glycerol. The objective was to enhance YO and β-carotene co-production in Rhodosporidium toruloides through the application of individual and combined inducers at both flask and bioreactor scales. Among the individual inducers, 1 % w/v Tween 20 (TW) and 10 ppm ethephon (EP) significantly increased total yeast oil (TO) and total β-carotene (TC) concentrations, respectively. When TW and EP were used together, TO and TC production increased by 2.0 and 2.6-fold, respectively in the bioreactor compared to the flask. The YO primarily consisted of C16 and C18 long-chain fatty acids, and the β-carotene produced showed functional similarities to commercial β-carotene. This research highlights the potential of biodiesel waste as a sustainable feedstock for co-producing YO and β-carotene, with the dual-inducer strategy providing a simple and effective method for enhancing production efficiency.