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Securing new sources of renewable energy and achieving national self-sufficiency in natural gas have become increasingly important in recent times. The study described in this paper focuses on three geologically diverse underground gas reservoirs (UGS) that are the natural habitat of methane-producing archaea, as well as other microorganisms with which methanogens have various ecological relationships. The objective of this research was to describe the microbial metabolism of methane in these specific anoxic environments during the year. DNA sequencing analyses revealed the presence of different methanogenic communities and their metabolic potential in all sites studied. Hydrogenotrophic Methanobacterium sp. prevailed in Lobodice UGS, members of the hydrogenotrophic order Methanomicrobiales predominated in Dolní Dunajovice UGS and thermophilic hydrogenotrophic members of the Methanothermobacter sp. were prevalent in Tvrdonice UGS. Gas composition and isotope analyses were performed simultaneously. The results suggest that the biotechnological potential of UGS for biomethane production cannot be neglected.

The microbial population present in stratal water withdrawn from an underground town gas reservoir transformed H2 and CO2, which are the obligatory constituents of town gas, to methane. By a common procedure one type of mesophilic Gram‐positive methanogenic bacteria was enriched. The bacteria grew and produced methane with H2+ CO2 only. This finding suggests that methanogenic bacteria present in the underground town gas reservoir could be responsible for the consumption of hydrogen and carbon dioxide from the stored town gas, causing a diminution of the gas volume and an enrichment of the gas with methane. An isotropic analysis of the stored town gas, in which °13C for methane around −80% was found, supports this suggestion.

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