For batch cultivation, the optimal medium for the carotenoid hyperproducing Phaffia rhodozyma mutant HT-5FO1C contained 25 g/l glucose, 4 g/l yeast extract, 1 g/l urea, and 0.3 g/l monosodium phosphate. In the absence of urea or monosodium phosphate, carotenoid production of P. rhodozyma was decreased. Forty four g/l of biomass and 1.1 mg/g yeast of total carotenoids were obtained by supplying the feed medium containing 6.8 g/l urea and 80 g/l glucose.