Enhanced biomass production through a repeated sequential auto-and heterotrophic culture mode in Chlorella protothecoides

A repeated sequential auto-and heterotrophic (RSAH) culture mode was designed to enhance biomass of Chlorella protothecoides. Based on the result that the photosynthesis system may receive damage if the light period is more than 16 h, autotrophy was applied in the 16 h of the light cycle and mixotrophy using acetic acid and glucose in the 8 h of dark cycle. In the dark cycle, an organic carbon source was added according to the Monod equation to maintain activation of the TCA cycle and organic carbon source-to-cell conversion. When acetic acid and glucose were used as organic carbon sources, this culture method was found to be 32.3% and 12.6% higher in biomass, 2.59 and 2.67 times higher in the organic carbon source-to-cell conversion factor, and 2.17 and 2.32 times higher in ATP/ADP ratio, respectively, compared to mixotrophy. Through this new culture method, economic feasibility and carbon reduction capabilities in large-scale cultures can be achieved.