Gas Substrate Effects on Hydrogenotrophic Biomethanation in Flocculent and Granular Sludge Systems

The biotechnological conversion of CO 2 to biomethane represents an energy-efficient, environmentally friendly, and sustainable approach within the waste-to-energy cycle. This process, in which CO 2 and H 2 are converted to biomethane in anaerobic bioreactors, is referred to as hydrogenotrophic biomethane production. While several studies have investigated hydrogenotrophic biomethane production, there is a lack of research comparing flocculent and granular sludge inoculum in continuously operated systems fed with a gas substrate. Both granular and flocculent sludge possess distinct advantages: granular sludge offers higher density, stronger microbial cohesion, and superior settling performance, whereas flocculent sludge provides faster substrate accessibility and more rapid initial microbial activity. In this study, two UASB (Upflow Anaerobic Sludge Blanket) reactors operated under mesophilic conditions were continuously fed with synthetic off-gas composed of pure H 2 and CO 2 in a 4:1 ratio and were compared in terms of microbial community shifts and their effects on hydrogenotrophic biomethane production. Biomethane production reached 75 ± 2% in the granular sludge reactor, significantly higher than the 64 ± 1.3% obtained with flocculent sludge. Although hydrogen consumption did not differ significantly, the granular sludge reactor exhibited higher CO 2 removal efficiency. Microbial analyses further revealed that granular sludge was more effective in supporting methanogenic archaea under conditions of gas substrate feeding. These findings offer advantageous suggestions for improving biogas production, enhancing waste gas management, and advancing sustainable energy generation.