Worldwide, mine gas is being used increasingly for heat and power production. About 7% of the annual methane emissions originate from coal mining. In abandoned coal mines, stable carbon and hydrogen isotopic signatures of methane indicate a mixed thermogenic and biogenic origin. The thermogenic methane is a reminder of geological processes, but its biogenic formation is still going on. Besides hard coal, possible sources for methane are large amounts of mine timber left behind after the end of mining.
Methanogenic archaea are responsible for the production of substantial amounts of methane. Mine timber and hard coal showed an in situ production of methane with isotopic signatures similar to those of the methane in the mine atmosphere. Long-term incubations of coal and timber as sole carbon sources formed methane over a period of 9 months. We directly unraveled the active methanogens mediating the methane release as well as the active bacteria potentially involved in the trophic network. Furthermore, we proved the presence of an active methanotrophic community. Directed by the methane production and oxidation, respectively, samples for DNA stable-isotope probing (SIP) coupled to subsequent quantitative PCR and DGGE analyses were taken from long term incubations over 6 months. The stable-isotope-labeled precursors of methane, [13C]acetate and H2-13CO2, and 13CH4 were fed to liquid cultures from hard coal and mine timber. Predominantly acetoclastic methanogenesis was stimulated in enrichments containing acetate and H2+CO2. The H2+CO2 was mainly used by acetogens similar to Pelobacter acetylenicus and Clostridium species forming acetate as intermediate and providing it to the methanogens. Active methanogens, closely affiliated to Methanosarcina barkeri, utilized the readily available acetate rather than the thermodynamical more favourable hydrogen. Furthermore, the activity of a distinct methane-oxidizing community is predominated of a member belonging to the type I methanotrophs similar to Methylobacter marinus that assimilated 13CH4 nearly exclusively. Thus, active methanotrophic bacteria are associated with the methanogenic microbial community that is highly adapted to the low H2 conditions found in the coal mines with acetate as the main precursor of the biogenic methane.