Effective Two-Stage Heterotrophic Cultivation of the Unicellular Green Microalga Chromochloris zofingiensis Enabled Ultrahigh Biomass and Astaxanthin Production

Chromochloris zofingiensis has obtained particular interest as a promising candidate for natural astaxanthin production. In this study, we established a two-stage heterotrophic cultivation process, by using which both the growth of C. zofingiensis and astaxanthin accumulation are substantially enhanced. Specifically, the ultrahigh biomass concentration of 221.3 g L-1 was achieved under the optimum culture conditions in 7.5 L fermenter during 12 days. When scaled-up in the 500 L fermentor, the biomass yield reached 182.3 g L-1 in 9 days, while the astaxanthin content was 0.068% of DW. To further promote astaxanthin accumulation, gibberellic Acid-3 (GA3) was screened from a variety of phytohormones and was combined with increased C/N ratio and NaCl concentration for induction. When C. zofingiensis was grown with the two-stage cultivation strategy, the astaxanthin yield reached 0.318 g L-1, of which the biomass yield was 235.4 g L-1 and astaxanthin content was 0.144% of DW. The content of the total fatty acids increased from 23 to 42% of DW simultaneously. Such an astaxanthin yield was 5.4-fold higher than the reported highest record and surpassed the level of Haematococcus pluvialis. This study demonstrated that heterotrophic cultivation of C. zofingiensis is competitive for industrial astaxanthin production.

Automated summary

This study established a two-stage heterotrophic cultivation process to enhance the growth and astaxanthin accumulation of Chromochloris zofingiensis. By optimizing culture conditions and adding gibberellic Acid-3 (GA3) as well as increasing the C/N ratio and NaCl concentration, the researchers achieved high biomass and astaxanthin yield, surpassing the previous records.