Pilot-scale production of selenium-enriched Nostoc sphaeroides colonies and polysaccharides using two-phase cultivation strategy

Nostoc sphaeroides , originally discovered in selenium-rich fields, is a photosynthetic, nitrogen-fixing cyanobacterium that forms edible spherical macrocolonies. However, prolonged selenium supplementation negatively impacts colony quality in culture. To address this, a two-phase cultivation strategy was developed to spatiotemporally separate biomass accumulation from selenium enrichment, resulting in high-quality selenium-enriched N. sphaeroides colonies. The first phase focused on colony growth in nitrogen-free BG-11 0 medium, while the second phase emphasized selenium enrichment in selenium-supplemented, phosphorus-reduced, and nitrogen-containing BG-11 medium (“selenium enrichment medium”). Scale-up to 300 L confirmed the robustness of this process, achieving desired colony hardness of ∼3 newtons, selenium content of ∼350 μg/g, and yield of ∼0.5 g/L (dry weight). Additionally, polysaccharides containing ∼110 μg/g selenium were isolated from the colonies, exhibiting biochemical properties similar to selenium-free polysaccharides from control colonies. These results provide valuable insight into the potential of N. sphaeroides as a cell factory chassis for producing selenium bioproducts.