The industrialized production of microbial lipids and carotenoids is facing various challenges including difficulties with microbial strains, fermentation processes, and downstream processing. This study aimed to optimize the fermentation conditions for the efficient co-production of lipids and carotenoids using Rhodosporidium toruloides. By using temperature and pH fermentation shift strategies, the lipid content was increased by 0.57 times (78.6%), and the carotenoid content was increased by 2.5 times (2.71 mg g −1 ) in comparison with a control. To reduce the fermentation cost, an inexpensive carbon source, cane molasses, was used to replace nitrogen, phosphorus, and carbon sources. This enabled a maximum lipid content of 68.9% and 2.85 mg g −1 of carotenoids to be obtained. These results provided an efficient method for lipid and carotenoid co-production, which could provide a basis for the industrial production of microbial lipids and carotenoids.