Abstract:The aim of this study was to promote fucoxanthin accumulation in Phaeodactylum tricornutum by photo-fermentation through optimizing the mode of multiple nitrogen supplementation and blue light enhancement. The results showed that the mixed nitrogen source (tryptone:urea=1:1, N mol/N mol; total nitrogen concentration at 0.02 mol/L) added to the culture system by six times was the best mode in shake flasks. Two-phase culture with light adjustment was then carried out in 5 L photo-fermenter with an enhanced blue light (R:G:B=67.1:16.7:16.3) in the second phase, leading to improved cell density (1.12×108 cells/mL), biomass productivity (330 mg/(d·L)), fucoxanthin content (19.62 mg/g), titer (69.71 mg/L) and productivity (6.97 mg/(d·L)). Compared with one-phase culture under red/blue (R:G:B=70.9:18.3:10.9) light and six-times nitrogen supplementation, the fucoxanthin content was significantly increased by 7.68% (P<0.05) but the productivity did not change significantly (P>0.05). Compared with one-phase culture under red/blue (R:G:B=70.9:18.3:10.9) light and one-time nitrogen supplementation, the content and productivity of fucoxanthin were significantly increased by 45.98% and 48.30% (P<0.05), respectively. This study developed a two-phase culture mode with multiple nitrogen supplementation and blue light enhancement, which effectively promoted the accumulation of fucoxanthin and improved the efficiency of nitrogen source utilization, thus providing a new approach for fucoxanthin accumulation in P. tricornutum by photo-fermentation.