Methane production from coal by a single methanogen.

Coal-bed methane is one of the largest unconventional natural gas resources. Although microbial activity may greatly contribute to coal-bed methane formation, it is unclear whether the complex aromatic organic compounds present in coal can be used for methanogenesis. We show that deep subsurface-derived Methermicoccus methanogens can produce methane from more than 30 types of methoxylated aromatic compounds (MACs) as well as from coals containing MACs. In contrast to known methanogenesis pathways involving one- and two-carbon compounds, this "methoxydotrophic" mode of methanogenesis couples O-demethylation, CO2 reduction, and possibly acetyl-coenzyme A metabolism. Because MACs derived from lignin may occur widely in subsurface sediments, methoxydotrophic methanogenesis would play an important role in the formation of natural gas not limited to coal-bed methane and in the global carbon cycle.

[Methane-generating potential of coal samples with different maturity].

OBJECTIVE: To evaluate coal bed methane production potential and characterize the in situ microbial communities of coal bed. METHODS: Coal samples were incubated under anaerobic conditions: mimicking coal bed condition, supplementing with methanogenic hydrocarbon degrading consortium, or adding with exogenetic substrate. Methane production was observed over time using gas chromatograph, and the in situ bacterial and archaeal communities were revealed using pyrosequencing. RESULTS: Enrichment incubation revealed that 3 of total 10 coal samples microcosms produced methane; bioaugmentation and substrate addition could enhance methane production of coal sample HF. Hydrogenotrophic Methanoculleus and acetoclastic Methanosaeta dominated the archaeal community of coal sample SL, while the bacterial domain was mainly composed of Firmicutes (54.4%), Proteobacteria (30.9%), uncultured bacteria (10.8%), Caldiserica (1.5%) and Thermotogae (1.3%). CONCLUSION: The methane production potential of coal bed samples with different maturity is different; the in situ coal bed microcosms are likely involved in hydrocarbons degradation and methane production.

The role of a plant/fungal consortium in the degradation of bituminous hard coal.

The sporadic growth of Cynodon dactylon was observed to occur directly on the surface of hard coal in dumps of the Witbank coal mining area of South Africa with the surface coal being broken down into a humic-like particulate material. Microorganism analysis of plants and rhizosphere material from the dumps revealed the presence of arbuscular mycorrhizal fungi and the coal solubilising fungus, Neosartorya fischeri. Studies established to replicate the dump environment revealed increased coal degradation in the form of humic acid production and an increase in small size particles as a result of Cynodon dactylon growth in association with arbuscular mycorrhizal fungi and Neosartorya fischeri. Results suggest that interactions between Cynodon dactylon, arbuscular mycorrhizal fungi, Neosartorya fischeri and other coal-degrading rhizosphere fungi could lead to the degradation of hard coal in situ and that the application of these organisms to discard dumps could be a novel method of coal dump rehabilitation.

Stimulation of methane generation from nonproductive coal by addition of nutrients or a microbial consortium.

Biogenic formation of methane from coal is of great interest as an underexploited source of clean energy. The goal of some coal bed producers is to extend coal bed methane productivity and to utilize hydrocarbon wastes such as coal slurry to generate new methane. However, the process and factors controlling the process, and thus ways to stimulate it, are poorly understood. Subbituminous coal from a nonproductive well in south Texas was stimulated to produce methane in microcosms when the native population was supplemented with nutrients (biostimulation) or when nutrients and a consortium of bacteria and methanogens enriched from wetland sediment were added (bioaugmentation). The native population enriched by nutrient addition included Pseudomonas spp., Veillonellaceae, and Methanosarcina barkeri. The bioaugmented microcosm generated methane more rapidly and to a higher concentration than the biostimulated microcosm. Dissolved organics, including long-chain fatty acids, single-ring aromatics, and long-chain alkanes accumulated in the first 39 days of the bioaugmented microcosm and were then degraded, accompanied by generation of methane. The bioaugmented microcosm was dominated by Geobacter sp., and most of the methane generation was associated with growth of Methanosaeta concilii. The ability of the bioaugmentation culture to produce methane from coal intermediates was confirmed in incubations of culture with representative organic compounds. This study indicates that methane production could be stimulated at the nonproductive field site and that low microbial biomass may be limiting in situ methane generation. In addition, the microcosm study suggests that the pathway for generating methane from coal involves complex microbial partnerships.

Phyto-bioconversion of hard coal in the Cynodon dactylon/coal rhizosphere.

Fundamental processes involved in the microbial degradation of coal and its derivatives have been well documented. A mutualistic interaction between plant roots and certain microorganisms to aid growth of plants such as Cynodon dactylon (Bermuda grass) on hard coal dumps has recently been suggested. In the present study coal bioconversion activity of nonmycorrhizal fungi was investigated in the C. dactylon/coal rhizosphere. Fungal growth on 2% Duff-agar, gutation formation on nitric acid treated coal and submerged culture activity in nitrogen-rich and -deficient broth formed part of the screening and selection of the fungi. The selected fungal isolates were confirmed to be found in pristine C. dactylon/coal rhizosphere. To simulate bioconversion, a fungal aliquot of this rhizosphere was used as inoculum for a Perfusate fixed bed bioreactor, packed with coal. The results demonstrate an enhanced coal bioconversion facilitated by low molecular weight organics and the bioconversion of coal may be initiated by an introduction of nitrogen moieties to the coal substrate. These findings suggest a phyto-bioconversion of hard coal involving plant and microbes occurring in the rhizosphere to promote the growth of C. dactylon. An understanding of this relationship can serve as a benchmark for coal dumps rehabilitation as well as for the industrial scale bioprocessing of hard coal.

Fungal biodegradation of hard coal by a newly reported isolate, Neosartorya fischeri.

Cynodon dactylon (Bermuda grass) has been observed to grow sporadically on the surface of coal dumps in the Witbank coal mining area of South Africa. Root zone investigation indicated that a number of fungal species may be actively involved in the biodegradation of hard coal, thus enabling the survival of the plant, through mutualistic interaction, in this extreme environment. In an extensive screening program of over two thousand samples, the Deuteromycete, Neosartorya fischeri, was isolated and identified. The biodegradation of coal by N. fischeri was tested in flask studies and in a perfusion fixed-bed bioreactor used to simulate the coal dump environment. The performance of N. fischeri was compared to Phanaerochaete chrysosporium and Trametes (Polyporus) versicolor, previously described in coal biodegradation studies. Fourier transform infrared spectrometry and pyrolysis gas chromatography mass spectrometry of the biodegradation product indicated oxidation of the coal surface and nitration of the condensed aromatic structures of the coal macromolecule as possible reaction mechanisms in N. fischeri coal biodegradation. This is a first report of N. fischeri-mediated coal biodegradation and, in addition to possible applications in coal biotechnology, the findings may enable development of sustainable technologies in coal mine rehabilitation.

Degradation of low rank coal by Trichoderma atroviride ES11.

A new isolate of Trichoderma atroviride has been shown to grow on low rank coal as the sole carbon source. T. atroviride ES11 degrades approximately 82% of particulate coal (10 g l(-1)) over a period of 21 days with 50% reduction in 6 days. Glucose (5 g l(-1)) as a supplemented carbon source enhanced the coal solubilisation efficiency of T. atroviride ES11, while 10 and 20 g l(-1) glucose decrease coal solubilisation efficiency. Addition of nitrogen [1 g l(-1) (NH(4))(2)SO(4)] to the medium also increased the coal solubilisation efficiency of T. atroviride ES11. Assay results from coal-free and coal-supplemented cultures suggested that several intracellular enzymes are possibly involved in coal depolymerisation processes some of which are constitutive (phenol hydroxylase) and others that were activated or induced in the presence of coal (2,3-dihydrobiphenyl-2,3-diol dehydrogenase, 3,4-dihydro phenanthrene-3,4-diol dehydrogenase, 1,2-dihydro-1,2-dihydroxynaphthalene dehydrogenase, 1,2-dihydro-1,2-dihydroxyanthracene dehydrogenase). GC-MS analysis of chloroform extracts obtained from coal degrading T. atroviride ES11 cultures showed the formation of only a limited number of specific compounds (4-hydroxyphenylethanol, 1,2-benzenediol, 2-octenoic acid), strongly suggesting that the intimate association between coal particles and fungal mycelia results in rapid and near-quantitative transfer of coal depolymerisation products into the cell.