Draft Genome Sequence of Methanobacterium sp. Maddingley, Reconstructed from Metagenomic Sequencing of a Methanogenic Microbial Consortium Enriched from Coal-Seam Gas Formation Water.
ABSTRACT: The draft genome of Methanobacterium sp. Maddingley was reconstructed from metagenomic sequencing of a methanogenic microbial consortium enriched from coal-seam gas formation water. It is a hydrogenotrophic methanogen predicted to grow using hydrogen and carbon dioxide.
Project description:Biogenic and biogenic-thermogenic coalbed methane (CBM) are important energy reserves for unconventional natural gas. Thus, to investigate biogenic gas formation mechanisms, a series of fresh coal samples from several representative areas of China were analyzed to detect hydrogen-producing bacteria and methanogens in an in situ coal seam. Complete microbial DNA sequences were extracted from enrichment cultures grown on coal using the Miseq high-throughput sequencing technique to study the diversity of microbial communities. The species present and differences between the dominant hydrogen-producing bacteria and methanogens in the coal seam are then considered based on environmental factors.Sequences in the Archaea domain were classified into four phyla and included members from Euryarchaeota, Thaumarchaeota, Woesearchaeota, and Pacearchaeota. The Bacteria domain included members of the phyla: Firmicutes, Proteobacteria, Bacteroidetes, Actinobacteria, Acidobacteria, Verrucomicrobia, Planctomycetes, Chloroflexi, and Nitrospirae. The hydrogen-producing bacteria was dominated by the genera: Clostridium, Enterobacter, Klebsiella, Citrobacter, and Bacillus; the methanogens included the genera: Methanorix, Methanosarcina, Methanoculleus, Methanobrevibacter, Methanobacterium, Methanofollis, and Methanomassiliicoccus.Traces of hydrogen-producing bacteria and methanogens were detected in both biogenic and non-biogenic CBM areas. The diversity and abundance of bacteria in the biogenic CBM areas are relatively higher than in the areas without biogenic CBM. The community structure and distribution characteristics depend on coal rank, trace metal elements, temperature, depth and groundwater dynamic conditions. Biogenic gas was mainly composed of hydrogen and methane, the difference and diversity were caused by microbe-specific fermentation of substrates; as well as by the environmental conditions. This discovery is a significant contribution to extreme microbiology, and thus lays the foundation for research on biogenic CBM.
Project description:Coal seam degasification and its efficiency are directly related to the safety of coal mining. Degasification activities in the Black Warrior basin started in the early 1980s by using vertical boreholes. Although the Blue Creek seam, which is part of the Mary Lee coal group, has been the main seam of interest for coal mining, vertical wellbores have also been completed in the Pratt, Mary Lee, and Black Creek coal groups of the Upper Pottsville formation to degasify multiple seams. Currently, the Blue Creek seam is further degasified 2-3 years in advance of mining using in-seam horizontal boreholes to ensure safe mining. The studied location in this work is located between Tuscaloosa and Jefferson counties in Alabama and was degasified using 81 vertical boreholes, some of which are still active. When the current long mine expanded its operation into this area in 2009, horizontal boreholes were also drilled in advance of mining for further degasification of only the Blue Creek seam to ensure a safe and a productive operation. This paper presents an integrated study and a methodology to combine history matching results from vertical boreholes with production modeling of horizontal boreholes using geostatistical simulation to evaluate spatial effectiveness of in-seam boreholes in reducing gas-in-place (GIP). Results in this study showed that in-seam wells' boreholes had an estimated effective drainage area of 2050 acres with cumulative production of 604 MMscf methane during ~2 years of operation. With horizontal borehole production, GIP in the Blue Creek seam decreased from an average of 1.52 MMscf to 1.23 MMscf per acre. It was also shown that effective gas flow capacity, which was independently modeled using vertical borehole data, affected horizontal borehole production. GIP and effective gas flow capacity of coal seam gas were also used to predict remaining gas potential for the Blue Creek seam.
Project description:A draft genome sequence of Methanobacterium sp. strain 34x was reconstructed from the metagenome of an enriched electromethanogenic biocathode operated in a microbial electrosynthesis (MES) reactor. Methanobacterium sp. strain 34x has 68.98% nucleotide-level genomic similarity with the closest related methanogen available with a whole-genome assembly, Methanobacterium lacus strain AL-21. This genome will provide insight into the functional potential of methanogens at the biocathodes of MES systems.
Project description:We present the high-quality draft genome of Methanobacterium subterraneum DF, a hydrogenotrophic methanogen that was isolated from deer feces. This organism has potentially been overlooked in previous studies. Interestingly, its genome encoded bile salt hydrolase, a crucial enzyme for bile salt tolerance that is found in gut organisms.
Project description:Background:The Xilingol grassland ecosystem has abundant superficial coal reserves. Opencast coal mining and burning of coal for electricity have caused a series of environmental challenges. Biogenic generation of methane from coal possesses the potential to improve economic and environmental outcomes of clean coal utilization. However, whether the microbes inhabiting the grassland soil have the functional potential to convert coal into biomethane is still unclear. Results:Microbial communities in an opencast coal mine and in grassland soil covering and surrounding this mine and their biomethane production potential were investigated by Hiseq sequencing and anaerobic cultivation. The microbial communities in covering soil showed high similarity to those in the surrounding soil, according to the pairwise weighted UniFrac distances matrix. The majority of bacterial communities in coal and soil samples belonged to the phyla Firmicutes, Bacteroidetes, Actinobacteria and Proteobacteria. The dominant bacterial genera in grassland soil included Gaiella, Solirubrobacter, Sphingomonas and Streptomyces; whereas, the most abundant genus in coal was Pseudarthrobacter. In soil, hydrogenotrophic Methanobacterium was the dominant methanogen, and this methanogen, along with acetoclastic Methanosarcina and methylotrophic Methanomassiliicoccus, was detected in coal. Network-like Venn diagram showed that an average of 28.7% of microbial communities in the samples belonged to shared genera, indicating that there is considerable microbial overlap between coal and soil samples. Potential degraders and methanogens in the soil efficiently stimulated methane formation from coal samples by the culturing-based approach. The maximum biogenic methane yields from coal degradation by the microbial community cultured from grassland soil reached 22.4 ?mol after 28 day. Conclusion:The potential microbial coal degraders and methanogenic archaea in grassland soil were highly diverse. Significant amounts of biomethane were generated from coal by the addition of grassland soil microbial communities. The unique species present in grassland soil may contribute to efficient methanogenic coal bioconversion. This discovery not only contributes to a better understanding of global microbial biodiversity in coal mine environments, but also makes a contribution to our knowledge of the synthetic microbiology with regard to effective methanogenic microbial consortia for coal degradation.
Project description:To solve the hidden danger of high methane and low permeability gas in the coal mining process, potentially affecting the safety production in an orderly way, we propose the use of deep hole blasting technology to improve the permeability of the coal seam gas drainage, increase the quantity and rate of extraction, and reduce methane output. Taking the geological conditions of the 201 working surface of Tingnan Coal Mine as an example, it is calculated that the single drilled fracture crack extension range is 3.11~5.24 m according to the coal seam deep-hole pre-splitting blasting joint mechanism and fracture propagation mechanics model, providing a theoretical basis for choosing the appropriate hole spacing. Using COMSOL simulation software to simulate the effective gas drainage radius of a coal seam from a two-dimensional perspective on a single borehole radial, the least squares fitting method was used to analyze the simulated data, and obtained the effective drilling extraction radius after pre-split blasting in a deep hole that is 3.6 m, which is in accordance with the theoretical calculations. In order to obtain accurate and scientific calculations, Fast lagrangian analysis of continua (FLAC3D) numerical simulation software was used. After simulating the distribution of plastic zone between two blast holes at different intervals from a three-dimensional angle, and evaluating the development of cracks in the blasting hole, the white zone of the blasting space was completely eliminated when the interval between blasting holes was 7 m, and the cracks could be propagated throughout the surroundings. Therefore, a blasting hole spacing of 7 m is optimal. On-site monitoring in the Nanting coal mine showed that the quantity and rate of extraction of the single hole after pre-splitting blasting were 2.36 times and 1.62 times as much as before. By integrating the borehole drainage amount and the optimized calculation equation, it could be concluded that the permeability coefficient of the coal seam after blasting was 7.78 times as much as before. The function of time-variated drilling methane emission was obtained using multivariate statistical regressions based on the on-site monitored borehole methane emission (qt), and the drilling limit after pre-splitting blasting revealed that the limitation of methane extraction volume was 5.27 times as much as before.
Project description:Despite the significance of biogenic methane generation in coal beds, there has never been a systematic long-term evaluation of the ecological response to biostimulation for enhanced methanogenesis in situ. Biostimulation tests in a gas-free coal seam were analysed over 1.5 years encompassing methane production, cell abundance, planktonic and surface associated community composition and chemical parameters of the coal formation water. Evidence is presented that sulfate reducing bacteria are energy limited whilst methanogenic archaea are nutrient limited. Methane production was highest in a nutrient amended well after an oxic preincubation phase to enhance coal biofragmentation (calcium peroxide amendment). Compound-specific isotope analyses indicated the predominance of acetoclastic methanogenesis. Acetoclastic methanogenic archaea of the Methanosaeta and Methanosarcina genera increased with methane concentration. Acetate was the main precursor for methanogenesis, however more acetate was consumed than methane produced in an acetate amended well. DNA stable isotope probing showed incorporation of 13C-labelled acetate into methanogenic archaea, Geobacter species and sulfate reducing bacteria. Community characterisation of coal surfaces confirmed that methanogenic archaea make up a substantial proportion of coal associated biofilm communities. Ultimately, methane production from a gas-free subbituminous coal seam was stimulated despite high concentrations of sulfate and sulfate-reducing bacteria in the coal formation water. These findings provide a new conceptual framework for understanding the coal reservoir biosphere.
Project description:Accumulation of carbon dioxide (CO2), associated with global temperature rise, and drastically decreasing fossil fuels necessitate the development of improved renewable and sustainable energy production processes. A possible route for CO2 recycling is to employ autotrophic and hydrogenotrophic methanogens for CO2-based biological methane (CH4) production (CO2-BMP). In this study, the physiology and productivity of Methanobacterium thermaggregans was investigated in fed-batch cultivation mode. It is shown that M. thermaggregans can be reproducibly adapted to high agitation speeds for an improved CH4 productivity. Moreover, inoculum size, sulfide feeding, pH, and temperature were optimized. Optimization of growth and CH4 productivity revealed that M. thermaggregans is a slightly alkaliphilic and thermophilic methanogen. Hitherto, it was only possible to grow seven autotrophic, hydrogenotrophic methanogenic strains in fed-batch cultivation mode. Here, we show that after a series of optimization and growth improvement attempts another methanogen, M. thermaggregas could be adapted to be grown in fed-batch cultivation mode to cell densities of up to 1.56 g L-1. Moreover, the CH4 evolution rate (MER) of M. thermaggregans was compared to Methanothermobacter marburgensis, the CO2-BMP model organism. Under optimized cultivation conditions, a maximum MER of 96.1?±?10.9 mmol L-1 h-1 was obtained with M. thermaggregans-97% of the maximum MER that was obtained utilizing M. marburgensis in a reference experiment. Therefore, M. thermaggregans can be regarded as a CH4 cell factory highly suited to be applicable for CO2-BMP.
Project description:Clostridium sp. Maddingley was isolated as an axenic culture from a brown coal-seam formation water sample collected from Victoria, Australia. It lacks the solventogenesis genes found in closely related clostridial strains. Metabolic reconstructions suggest that volatile fatty acids are the main fermentation end products.
Project description:Cellulosilyticum sp. strain I15G10I2 was isolated from a coal seam gas water treatment pond at the Spring Gully water treatment facility, Roma, Queensland, Australia. Analysis of the genome of 4,489,861 bp and G+C content of 35.23% revealed that strain I15G10I2 shared limited similarity to members of the genus Cellulosilyticum, family Lachnospiraceae.