Iron and nitrogen regulate carbon transformation in a methanotroph-microalgae system

Nutrient supply is important for maintaining a methanotroph and microalgae (MOB-MG) system for biogas valorization. However, there is a lack of understanding regarding how key elements regulate the growth of a MOB-MG coculture. In this study, a MOB-MG coculture with high protein content (0.47 g/g biomass) was established from waste activated sludge using synthetic biogas. An increase in iron availability substantially stimulated the specific growth rate (from 0.18 to 0.62 day⁻¹) and biogas conversion rate (from 26.81 to 106.57 mg-C L⁻¹ day⁻¹) of the coculture. Moreover, the protein content remained high (0.51 g/g biomass), and the total lipid content increased (from 0.09 to 0.14 g/g biomass). Nitrogen limitation apparently constrained the specific growth rate (from 0.64 to 0.28 day⁻¹) and largely reduced the protein content (from 0.51 to 0.31 g/g biomass) of the coculture. Intriguingly, the lipid content remained unchanged after nitrogen was depleted. The eukaryotic community was consistently dominated by MG belonging to Chlorella, while the populations of MOB shifted from Methylococcus/Methylosinus to Methylocystis due to iron and nitrogen amendment. In addition, diverse non-methanotrophic heterotrophs were present in the community. Their presence neither compromised the performance of the coculture system nor affected the protein content of the biomass. However, these heterotrophs may contribute to high carbon conversion efficiency by utilizing the dissolved organic carbon released by MOB and MG. Overall, the findings highlight the vital roles of iron and nitrogen in achieving efficient conversion of biogas, fast growth of cells, and optimal biomass composition in a MOB-MG coculture system.