An innovative multistage anaerobic hythane reactor (MAHR): Metabolic flux, thermodynamics and microbial functions

Biohythane production from wastewater via anaerobic fermentation currently relies on two-stage physically separated biohydrogen and biomethane reactors, which requires closed monitoring, the implementation of a control system, and cost-intensive, complex operation. Herein, an innovative multistage anaerobic hythane reactor (MAHR) was reported via integrating two-stage fermentation into one reactor. MAHR was constructed using an internal down-flow packed bed reactor and an external up-flow sludge blanket to enhance microbial enrichment and thermodynamic feasibility of the associated bioreactions. The performance of MAHR was investigated for 160 d based on biogas production, metabolic flux and microbial structure in comparison to a typical anaerobic high-rate reactor (up-flow anaerobic sludge blanket (UASB)). A biohythane production with an optimized hydrogen volume ratio (10-20%) and a high methane content (75-80%) was achieved in the hythane zone (M_H) and methane zone (M_M) in MAHR, respectively. In addition, MAHR showed a stronger capability to accommodate a high organic loading rate (120 g COD/L/d), and it enhanced the conversion of organics leading to a methane production rate 66% higher than UASB. Thermodynamic analysis suggested that hydrogen extraction in M_H significantly decreased the hydrogen partial pressure (<0.1% vol) which favored acetogenesis in M_M. Metabolic flux and microbial function analysis further supported the superior performance of MAHR over UASB, which was primarily attributed to enhanced acetogenesis and acetoclastic methanogenesis.