An iron corrosion-assisted H₂-supplying system: a culture method for methanogens and acetogens under low H₂ pressures

H₂ is an important fermentation intermediate in anaerobic environments. Although H₂ occurs at very low partial pressures in the environments, the culture and isolation of H₂-utilizing microorganisms is usually carried out under very high H₂ pressures, which might have hampered the discovery and understanding of microorganisms adapting to low H₂ environments. Here we constructed a culture system designated the "iron corrosion-assisted H₂-supplying (iCH) system" by connecting the gas phases of two vials (one for the iron corrosion reaction and the other for culturing microorganisms) to achieve cultures of microorganisms under low H₂ pressures. We conducted enrichment cultures for methanogens and acetogens using rice paddy field soil as the microbial source. In the enrichment culture of methanogens under canonical high H₂ pressures, only Methanobacterium spp. were enriched. By contrast, Methanocella spp. and Methanoculleus spp., methanogens adapting to low H₂ pressures, were specifically enriched in the iCH cultures. We also observed selective enrichment of acetogen species by the iCH system (Acetobacterium spp. and Sporomusa spp.), whereas Clostridium spp. predominated in the high H₂ cultures. These results demonstrate that the iCH system facilitates culture of anaerobic microorganisms under low H₂ pressures, which will enable the selective culture of microorganisms adapting to low H₂ environments.