Zeolitic imidazolate framework-derived porous carbon enhances methanogenesis by facilitating interspecies electron transfer: Understanding fluorimetric and electrochemical responses of multi-layered extracellular polymeric substances

Modulating microbial electron transfer during anaerobic digestion can significantly improve syntrophic interactions for enhanced biogas production. As a carbonaceous conductive material, zeolite imidazolate framework-67 (ZIF-67)-derived porous carbon (PC) was hypothesized to act as a microbial electron transfer highway and assessed with respect to understanding the fluorimetric and electrochemical responses of multilayered extracellular polymeric substances (EPS). The highest biomethane yield (614.0 mL/g) from ethanol was achieved in the presence of 100 mg/L PC prepared at a carbonization temperature of 800 °C (PC-800), which was 28.2% higher than that without PC addition. Electrochemical analysis revealed that both the redox peak currents and conductivity of the methanogenic sludge increased, while the free charge transfer resistance decreased with PC-800 addition. The conductive PC-800 potentially functioned as an abiotic electron conduit to promote direct interspecies electron transfer, thereby resulting in decreased expression of functional genes associated with electrically conductive pili (e-pili) and hemeproteins. Additionally, PC-800 stimulated the secretion of redox-active humic substances (HSs), and excitation emission matrix spectra analysis indicated that the largest increase in percent fluorescence response of HSs occurred in the tightly bound EPS (TB-EPS) with addition of PC-800. This was attributed to the strong complexation ability of PC-800 particles to hydroxyl/carboxylic/phenolic moieties of HSs contained in the TB-EPS. Microbial analysis revealed that syntrophic/exoelectrogenic bacteria such as Pelotomaculum and Syntrophomonas, as well as hydrogenotrophic/electrotrophic methanogens such as Methanoculleus and Methanobacterium, were enriched in methanogenic sludge with adding PC-800. This study provided comprehensive insights for understanding the interactions among ZIF-derived PC, methanogenic microorganisms and their multilayered EPS.