Significance of carbon to nitrogen ratio on the long-term stability of continuous photofermentative hydrogen production

The stable and optimized operation of photobioreactors (PBRs) is the most challenging task in photofermentative biological hydrogen production. The carbon to nitrogen ratio (C/N) in the feed is a critical parameter that significantly influences microbial growth and hydrogen production. In this study, the effects of changing the C/N ratio to achieve stable biomass and continuous hydrogen production using fed-batch cultures of Rhodobacter capsulatus YO3 (uptake hydrogenase deleted, hup-) were investigated. The experiments were carried out in 8 L panel PBRs operated in indoor conditions under continuous illumination and controlled temperature. Culture media containing different acetate (40–80 mM) and glutamate (2–4 mM) concentrations were used to study the effects of changing the C/N ratio on biomass growth and hydrogen production. Stable biomass concentration of 0.40 g dry cell weight per liter culture (gDCW/Lc) and maximum hydrogen productivity of 0.66 mmol hydrogen per liter culture per hour (mmol/Lc/h) were achieved during fed-batch operation with media containing 40 mM acetate and 4 mM glutamate, C/N = 25, for a period of over 20 days. A study on the effect of biomass recycling on biomass growth and hydrogen production showed that the feedback of cells into the photobioreactor improved biomass stability during the fed-batch operation but decreased hydrogen productivity.