In this paper,litchi processing residues was fermented with the lipid-producing rhodotorula glutinis to produce microbial lipid.The optimal seeding time and volume of rhodotorula glutinis seed broth in flask were studied.Meanwhile,the process of microbial oil production was optimized by single factor experiment and orthogonal experiment to obtain the optimal fermentation conditions,and the physicochemical properties of the oil were analyzed.Results showed that the capacity of lipid production of 378.3 mg/L was achieved under the optimal conditions as follows:24 h of seeding time,40 mL/250 of liquid volume in flask,8°Brix of initial sugar concentration,5.5 of initial pH,10％ of inoculum,and 96 h of fermentation time.Meanwhile,the biomass reached 15.670 g/L and the oil content was 2.414％.The microbial oil was yellow transparent liquid with average relative density of 1.104 4 g/L and refractive index of 1.459 0.Its acid value was 5.39 (mg/g),the iodine value was 101.8 (gI2/100 g),and the peroxide value was 9.44 (mmol/kg).The production of microbial oil with litchi residue reached the dual purpose of waste utilization and energy development and had the broad application prospect.