Chlorella sorokiniana has received particular attention as a promising candidate for microalgal biomass and lutein production. In this work, heterotrophic cultivation was explored to improve the lutein production efficiency of a lutein-rich microalga C. sorokiniana FZU60. Flask cultivation results showed that the highest lutein productivity was achieved at 30°C with an initial cell concentration of 1.40 g/L. Furthermore, six types of fed-batch strategies based on nutrient composition and concentration were examined using a 5 L fermenter. Among them, ultra-high lutein production (415.93 mg/L) and productivity (82.50 mg/L/d) with lutein content of 2.57 mg/g were achieved with fed-batch 3F (i.e., pulse-feeding with concentrated urea-N medium to achieve a 3-fold nutrient concentration). The lutein production performance achieved is much higher than the reported values. This work demonstrates that heterotrophic cultivation of C. sorokiniana FZU60 with the proposed fed-batch strategy could significantly enhance the production performance and the commercial viability of microalgae-derived lutein.