In a deep aquifer associated with an accretionary prism, significant methane (CH4) is produced by a subterranean microbial community. Here, we developed bioreactors for producing CH4 and hydrogen (H-2) using anaerobic groundwater collected from the deep aquifer. To generate CH4, the anaerobic groundwater amended with organic substrates was incubated in the bioreactor. At first, H-2 was detected and accumulated in the gas phase of the bioreactor. After the H-2 decreased, rapid CH4 production was observed. Phylogenetic analysis targeting 16S rRNA genes revealed that the H-2-producing fermentative bacterium and hydrogenotrophic methanogen were predominant in the reactor. The results suggested that syntrophic biodegradation of organic substrates by the H-2-producing fermentative bacterium and the hydrogenotrophic methanogen contributed to the CH4 production. For H-2 production, the anaerobic groundwater, amended with organic substrates and an inhibitor of methanogens (2-bromoethanesulfonate), was incubated in a bioreactor. After incubation for 24h, H-2 was detected from the gas phase of the bioreactor and accumulated. Bacterial 16S rRNA gene analysis suggested the dominance of the H-2-producing fermentative bacterium in the reactor. Our study demonstrated a simple and rapid CH4 and H-2 production utilizing anaerobic groundwater containing an active subterranean microbial community.
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