Hydrogen-entangled electron transfer is verified as an important extracellular pathway of sharing reducing equivalents to regulate biofilm activities within the diversely anaerobic environment, especially in microbial electrosynthesis system. However, the lack of useful methods for in-situ hydrogen detection in cathodic biofilm, the role of hydrogen involved in electron transfer is still debatable. Herein, the cathodic biofilm was constructed in the CH4-produced microbial electrosynthesis reactors, in which the hydrogen evolution dynamic was analyzed to confirm the presence of hydrogen-associated electron transfer nearby the cathode within the micrometre scale. The fluorescent in-situ hybridization images indicated that a colocalized community of archaea and bacteria developed within the thick of biofilm 58.10 μm at cathode suggesting the hydrogen gradient detected by microsensor was consumed by the collaboration of bacteria and archaea. Coupling microsensor and cyclic voltammetry test further provided the semi-quantitative results of hydrogen-associated contribution to methane generation ( around 21.20% ± 1.57% having a potential of -0.5 V to 0.69 V). This finding provides deep insight into the mechanism of electron transfer in biofilm on conductive materials.
IMPORTANCE: Electron transfer from the electrode to biofilm is of great interest to the fields of microbial electrochemical technology, bioremediation and methanogenesis. It has promising application potential to boost more value-added products or pollutant degradation. Importantly, the microbial ability to obtain electrons from electrode and utilizing it brings a new insight into the direct interspecies electron transfer during methanogenesis. Previous studies had verified the direct pathway of electron transfer from the electrode to the pure-culture bacterium, but it was rarely reported how the methanogenic biofilm of mixed cultures shares electrons by hydrogen-associated or hydrogen-free pathway. In the current study, a combination method of microsensor and cyclic voltammetry successfully semi-quantified the role of hydrogen in the electron transfer from the electrode to methanogenic biofilm.