Astaxanthin is a valuable carotenoid, which has been approved as a food coloring by the US Food and Drug Administration and is considered as a food dye by the European Union (European Commission). This work aimed to attain Rhodosporidium toruloides mutants for enhanced astaxanthin accumulation using ultraviolet (UV) and gamma irradiation mutagenesis. Gamma irradiation was shown to be more efficient than UV for producing astaxanthin-overproducer. Among the screened mutants, G17, a gamma-induced mutant, exhibited the highest astaxanthin production, which was significantly higher than that of the wild strain. Response surface methodology was then applied to optimize the medium compositions for maximizing astaxanthin production by the mutant G17. The optimal medium compositions for the cultivation of G17 were determined as a peptone concentration of 19.75 g/L, malt extract concentration of 13.56 g/L, and glucose concentration of 19.92 g/L, with the maximum astaxanthin yield of 3021.34 µg/L ± 16.49 µg/L. This study suggests that the R. toruloides mutant (G17) is a potential candidate for astaxanthin production.