Carotenoid compounds are the popular natural antioxidants which are often isolated from plants. There have been more and more researches on carotenoid biosynthesis towards lowering product prices. In this study, in order to produce carotenoid, Rhodotorula mucilaginosa was grown on aqueous media composed of carbon source (glucose, glycerol), nitrogen source (yeast extract, (NH4)2SO4). The optimum nutrient concentration was 10g/L glucose, 10g/L glycerol, the ratio of yeast extract and (NH4)2SO4 (3:7). The fermentation time for obtaining the highest carotenoid yield was 10 days in our research condition. Additionally, some oxidative stress environment for Rhodotorula mucilaginosa was be studied. The result has shown that the low level of Cu2+ (4.5mM) or 1% H2O2 solution (% v/v) in the fermentation media could increase the carotenoid biosynthesis.