Carboxylic acids production and electrosynthetic microbial community evolution under different CO₂ feeding regimens

Microbial electrosynthesis (MES) is a potential technology for CO₂ recycling, but insufficient information is available on the microbial interactions underpinning electrochemically-assisted reactions. In this study, a MES reactor was operated for 225 days alternately with bicarbonate or CO₂ as carbon source, under batch or continuous feeding regimens, to evaluate the response of the microbial communities, and their productivity, to dynamic operating conditions. A stable acetic acid production rate of 9.68 g m^-2 d^-1, and coulombic efficiency up to 40%, was achieved with continuous CO₂ sparging, higher than the rates obtained with bicarbonate (0.94 g m^-2 d^-1) and CO₂ under fed-batch conditions (2.54 g m^-2 d^-1). However, the highest butyric acid production rate (0.39 g m^-2 d^-1) was achieved with intermittent CO₂ sparging. The microbial community analyses focused on differential amplicon sequence variants (ASVs), allowing detection of ASVs significantly different across consecutive samples. This analysis, combined with co-occurence network analysis, and cyclic voltammetry, indicated that hydrogen-mediated acetogenesis was carried out by Clostridium, Eubacterium and Acetobacterium, whereas Oscillibacter and Caproiciproducens were involved in butyric acid production. The cathodic community was spatially inhomogeneous, with potential electrotrophs, such as Sulfurospirillum and Desulfovibrio, most prevalent near the current collector. The abundance of Sulfurospirillum positively correlated with that of Acetobacterium, supporting the syntrophic metabolism of both organisms.