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A detailed phylogeny for the Methanomicrobiales

The small subunit rRNA sequence of twenty archaea, members of the Methanomicrobiales, permits a detailed phylogenetic tree to be inferred for the group. The tree confirms earlier studies, based on far fewer sequences, in showing the group to be divided into two major clusters, temporarily designated the "methanosarcina" group and the "methanogenium" group. The tree also defines phylogenetic relationships within these two groups, which in some cases do not agree with the phylogenetic relationships implied by current taxonomic names--a problem most acute for the genus Methanogenium and its relatives. The present phylogenetic characterization provides the basis for a consistent taxonomic restructuring of this major methanogenic taxon.

NASA Discipline Number 52-30↗

The microbial community of Ophrydium versatile colonies: endosymbionts, residents, and tenants

Ophrydium versatile is a sessile peritrichous ciliate (Kingdom Protoctista, class Oligohymenophora, order Peritrichida, suborder Sessilina) that forms green, gelatinous colonies. Chlorophyll a and b impart a green color to Ophrydium masses due to 400-500 Chlorella-like endosymbionts in each peritrich. Ophrydium colonies, collected from two bog wetlands (Hawley and Leverett, Massachusetts) were analyzed for their gel inhabitants. Other protists include ciliates, mastigotes, euglenids, chlorophytes, and heliozoa. Routine constituents include from 50-100,000 Nitzschia per ml of gel and at least four other diatom genera (Navicula, Pinnularia, Gyrosigma, Cymbella) that may participate in synthesis of the gel matrix. Among the prokaryotes are filamentous and coccoid cyanobacteria, large rod-shaped bacteria, at least three types of spirochetes and one unidentified Saprospira-like organism. Endosymbiotic methanogenic bacteria, observed using fluorescence microscopy, were present in unidentified hypotrichous ciliates. Animals found inside the gel include rotifers, nematodes, and occasional copepods. The latter were observed in the water reservoir of larger Ophrydium masses. From 30-46% of incident visible radiation could be attenuated by Ophrydium green jelly masses in laboratory observations. Protargol staining was used to visualize the elongate macronuclei and small micronucleus of O. versatile zooids and symbiotic algal nuclei. Electron microscopic analysis of the wall of the Chlorella-like symbiont suggests that although the Ophrydium zooids from British Columbia harbor Chlorella vulgaris, those from Hawley Bog contain Graesiella sp. The growth habit in the photic zone and loose level of individuation of macroscopic Ophrydium masses are interpretable as extant analogs of certain Ediacaran biota: colonial protists in the Vendian fossil record.

Non-NASA Center↗

Predictive isotopic biogeochemistry: hydrocarbons from anoxic marine basins

Carbon isotopic compositions were determined for individual hydrocarbons in water column and sediment samples from the Cariaco Trench and Black Sea. In order to identify hydrocarbons derived from phytoplankton, the isotopic compositions expected for biomass of autotrophic organisms living in surface waters of both localities were calculated based on the concentrations of CO2(aq) and the isotopic compositions of dissolved inorganic carbon. These calculated values are compared to measured delta values for particulate organic carbon and for individual hydrocarbon compounds. Specifically, we find that lycopane is probably derived from phytoplankton and that diploptene is derived from the lipids of chemoautotrophs living above the oxic/anoxic boundary. Three acyclic isoprenoids that have been considered markers for methanogens, pentamethyleicosane and two hydrogenated squalenes, have different delta values and apparently do not derive from a common source. Based on the concentration profiles and isotopic compositions, the C31 and C33 n-alkanes and n-alkenes have a similar source, and both may have a planktonic origin. If so, previously assigned terrestrial origins of organic matter in some Black Sea sediments may be erroneous.

Non-NASA Center↗

Halotolerant and Resistant to High pH Hydrogenase from Haloalkaliphilic Sulfate-Reducing Bacterium Desulfonatronum thiodismutans

Hydrogenase is the key enzyme of energetic metabolism in cells, it catalyzing the converse reaction of hydrogen oxidation and responsible for consumption and excretion of hydrogen in bacteria. Hydrogenases are proteins containing either Nickel and Iron, or the only Iron in theirs active center. Hydrogenases have been found in many microorganisms, such as Methanogenic, acetogenic, nitrogen-fixing, photosynthetic and sulfate-reducing bacteria that could utilize the hydrogen as energy source or use it as electron sink. Hydrogenases are subject for wide physiological, biochemical, physicochemical and genetic studies due to theirs abilities produce the molecular hydrogen as alternative source of pure energy. Notwithstanding on enough large quantity of works that deal with intracellular and extrasellular enzymes of halophilic bacteria, the data about hydrogenases and theirs functions of salts practically are absent. The study of hydrogenase in cell-free extracts of extremely halophilic eubacterium Acetohalobium mabaticum showed dramatic increasing activity of the enzyme at high concentrations of NaCl and KCI (close to saturated solution). Here we present the data of free-cells extracted hydrogenase from new haloalkaliphilic sulfate-reducing bacterium Desulfonatronum thiodismutans, which grow on highly miniralized carbonate-bicarbonate medium in salinity range 1 to 7 % and at pH 7.8 - 10.5. Studied enzyme was active in Concentration range from 0 to 4.3 M NaCl with optimum at 1.0 M NaCl. At 1.0 M NaCl the enzyme activity was increased on 20 %, but with changing concentration from 2.1 M to 3.4 M the activity decreased and was kept on constant level. NaHCO3 inhibited hydrogenase activity on more then 30 %. The maximum of enzyme activity was observed at pH 9.5 with limits 7.5 and 11.5 that practically equal to pH optimum of bacterial growth. Therefore the hydrogenase of Desulfanatronum thiodismutans is tolerant to high concentrations of sodium salts and it also resistant to high pH that make it the unique subject for different biochemical research and detects the possibility for biotechnological application.

Detkova, Ekaterina N.↗

Carbon isotope effects associated with aceticlastic methanogenesis

The carbon isotope effects associated with synthesis of methane from acetate have been determined for Methanosarcina barkeri 227 and for methanogenic archaea in sediments of Wintergreen Lake, Michigan. At 37 degrees C, the 13C isotope effect for the reaction acetate (methyl carbon) --> methane, as measured in replicate experiments with M. barkeri, was - 21.3% +/- 0.3%. The isotope effect at the carboxyl portion of acetate was essentially equal, indicating participation of both positions in the rate-determining step, as expected for reactions catalyzed by carbon monoxide dehydrogenase. A similar isotope effect, - 19.2% +/- 0.3% was found for this reaction in the natural community (temperature = 20 degrees C). Given these observations, it has been possible to model the flow of carbon to methane within lake sediment communities and to account for carbon isotope compositions of evolving methane. Extension of the model allows interpretation of seasonal fluctuations in 13C contents of methane in other systems.

NASA Discipline Exobiology↗

Linking Archaeal Molecular Diversity and Lipid Biomarker Composition in a Hypersaline Microbial Mat Community

Lipid biomarkers for discrete microbial groups are a valuable tool for establishing links to ancient microbial ecosystems. Lipid biomarkers can establish organism source and function in contemporary microbial ecosystems (membrane lipids) and by analogy, potential relevance to the fossilized carbon skeletons (geolipids) extracted from ancient sedimentary rock. The Mars Exploration Rovers have provided clear evidence for an early wet Mars and the presence of hypersaline evaporitic basins. Ongoing work on an early Earth analog, the hypersaline benthic mats in Guerrero Negro, Baja California Sur, may provide clues to what may have evolved and flourished on an early wet Mars, if only for a short period. Cyanobacterial mats are a pertinent early Earth analog for consideration of evolutionary and microbial processes within the aerobic photosynthetic and adjacent anoxic layers. Fluctuations in physio-chemical parameters associated with spatial and temporal scales are expressed through vast microbial metabolic diversity. Our recent work hopes to establish the dynamic of archaeal diversity, particularly as it relates to methane production in this high sulfate environment, through the use of lipid biomarker and phylogenetic analyses. Archaeal 16s rRNA and mcrA gene assemblages, demonstrated distinct spatial separation over the 130 mm core of at least three distinct genera within the order Methanosarcinales, as well as an abundance of uncultured members of the Thermoplasmales and Crenarchaeota. Ether-bound lipid analysis identified abundant 0-alkyl and 0-isopranyl chains throughout the core, and the presence of sn-2 hydroxyarchaeol, a biomarker for methylotrophic methanogens. A unique ether isoprenoid chain, a C30:1 , possibly related to the geolipid squalane, a paleobiomarker associated with hypersaline environments, was most abundant within the oxic-anoxic transition zone.

Jahnke, Linda↗

Optical Detection of Life on Exoplanets

We describe what is known about the atmospheric properties (Teff, lob g, [FelH]) and fundamental properties (mass, age, and metal content) of nearby stars and how they influence the habitable zones and habitable eras of these stars. We then take an observer's point of view to assess the ability of optical telescopes to detect photosynthetic or methanogenic life on planets orbiting these stars.

Heap, Sara↗

Psychrophilic Biomass Producers in the Trophic Chain of the Microbial Community of Lake Untersee, Antarctica

The study of photosynthetic microorganisms from the Lake Untersee samples showed dispersed distribution of phototrophs within ~80 m water column. Lake Untersee represents a unique ecosystem that experienced complete isolation: sealed by the Anuchin Glacier for many millennia. Consequently, its biocenosis has evolved over a significant period of time without exchange or external interaction with species from other environments. The major producers of organic matter in Lake Untersee are represented by phototrophic and chemolithotrophic microorganisms. This is the traditional trophic scheme for lacustrine ecosystems on Earth. Among the phototrophs, diatoms were not found, which differentiates this lake from other known ecosystems. The dominant species among phototrophs was Chlamydomonas sp. with typical morphostructure: green chloroplasts, bright red round spot, and two polar flagella near the opening. As expected, the physiology of studied phototrophs was limited by low temperature, which defined them as obligate psychrophilic microorganisms. By the quantity estimation of methanogenesis in this lake, the litho-autotrophic production of organic matter is competitive with phototrophic production. However, pure cultures of methanogens have not yet been obtained. We discuss the primary producers of organic matter and the participation of our novel psychrophilic homoacetogen into the litho-autotrophic link of biomass production in Lake Untersee.

Pikuta, Elena V.↗

Possible Sources for Methane and C2-C5 Organics in the Plume of Enceladus

In this paper we consider six possible sources of CH4 and other low-mass (C2 - C5) organics in the plume of Enceladus: initial endowments of cometary organics or Titan- like tholin, in situ production by Fisher-Tropsch type reactions, water-rock reactions, or microbiology, and thermogenesis from heavier organics already present. We report on new laboratory results C2 hydrocarbons released on thermogenesis of laboratory tholin and Fisher-Tropsch type synthesis. Tholin heating produced ratios of CH4/C2H4 and CH4/C2H6 of about 2 for temperatures up to 450 C and about 6 for a temperature of 650 C. Low pressure Fisher-Tropsch type experiments produced CH4/C2H4 of approx 1.5, similar to previous results. No C2H2 was produced by either process. Tests of gas production by four strains of methanogens confirmed the absence of any detectable production of non-methane hydrocarbons. Cometary endowment, Fisher-Tropsch type synthesis, and Titan-like tholin incorporation could be primary inputs of organics and subsequent thermal processing of any of these all are possible sources of low mass organics in the plume. Biological production and water-rock reactions are an alternative source of CH4. Neither water-- ]rock reactions or thermal processing of biomass could be a source C2 . C5 organics due to the low interior pressures. The confirmed detection of CO and C2H2 in the plume of Enceladus would provide an important constraint on sources as we have identified no process . other than the initial volatile component of cometary organics which can supply these gases. Precise determination of the relative concentrations of C1 - C5 hydrocarbons may provide additional constraints on sources but a detailed isotopic analysis of C and H in these organics and a search for amino acids constitute the next important steps in resolving the sources of the organics in Enceladus' plume.

McKay, Chris↗

Modeling the Distribution and Type of High-Latitude Natural Wetlands for Methane Studies

High latitude (>50N) natural wetlands emit a substantial amount of methane to the atmosphere, and are located in a region of amplified warming. Northern hemisphere high latitudes are characterized by cold climates, extensive permafrost, poor drainage, short growing seasons, and slow decay rates. Under these conditions, organic carbon accumulates in the soil, sequestering CO2 from the atmosphere. Methanogens produce methane from this carbon reservoir, converting stored carbon into a powerful greenhouse gas. Methane emission from wetland ecosystems depends on vegetation type, climate characteristics (e.g, precipitation amount and seasonality, temperature, snow cover, etc.), and geophysical variables (e.g., permafrost, soil type, and landscape slope). To understand how wetland methane dynamics in this critical region will respond to climate change, we have to first understand how wetlands themselves will change and therefore, what the primary controllers of wetland distribution and type are. Understanding these relationships permits data-anchored, physically-based modeling of wetland distribution and type in other climate scenarios, such as paleoclimates or future climates, a necessary first step toward modeling wetland methane emissions in these scenarios. We investigate techniques and datasets for predicting the distribution and type of high latitude (>50N) natural wetlands from a suite of geophysical and climate predictors. Hierarchical clustering is used to derive an empirical methane-centric wetland model. The model is applied in a multistep process first to predict the distribution of wetlands from relevant geophysical parameters, and then, given the predicted wetland distribution, to classify the wetlands into methane-relevant types using an expanded suite of climate and biogeophysical variables. As the optimum set of predictor variables is not known a priori, the model is applied iteratively, and each simulation is evaluated with respect to observed high-latitude wetlands.

Romanski, Joy↗

Synthesis and oxidation of methane

The present disclosure describes genes and proteins of the coenzyme F430 synthetic pathway. The genes and proteins in the pathway find uses as isolated nucleic acids, transformation vectors, a transformation media, genetically modified cells, methods of modulating methanogenesis, methods of modulating methane oxidation, methods of making a tetrapyrrole compound, methods of oxidizing methane, methods of biogenic methane synthesis is provided, methods of assaying an organism for potential methanogenic or methanotrophic activity, and isolated proteins.

10 SYNTHETIC FUELS↗

BETO 2021 Peer Review - Biomethanation to Upgrade Biogas to Pipeline Grade Methane WBS 5.1.3.102

We are developing, innovating and de-risking a biomethanation process capable of megawatt-scale deployment that upgrades biogas waste streams to produce pipeline quality renewable natural gas (RNG). Biomethanation is a two-step process using a methanogenic microorganism to convert renewable hydrogen (H2) and waste carbon dioxide (CO2) to renewable methane (CH4) - the primary component in natural gas. Using biogenic CO2 from biogas sources like dairies, wastewater treatment plants, and landfills allows production of this drop-in direct replacement fuel to participate in the growing number of carbon markets; like California's Low Carbon Fuel Standard and the Federal Renewable Fuel Standard. The end-of-project goal is to demonstrate pipeline quality RNG production (> 95% CH4, < 4% H2, <1% CO2, < 0.2% O2 and < 4 parts per million hydrogen sulfide) using real biogas feedstocks. We will accomplish this goal by designing and building a pressurized (18 bar) mobile lab-scale (20L) bioreactor research platform, including integrated electrolyzer system, based on lessons learned from operating the 700L pilot system from Southern California Gas Company. In collaboration with Electrochaea, natural gas utilities and Argonne National lab, will provide the data to establish a preliminary range of carbon intensity to help accelerate the deployment of utility-scale H2 production and qualify the biomethanation pathway process for RNG production.

29 ENERGY PLANNING, POLICY, AND ECONOMY↗

Devices and methods for generating electrical current from methane

Methods, microbial fuel cells and microbial consortia for generating electrical current are provided according to the present invention which include providing a microbial consortium to an anode chamber of a microbial fuel cell, wherein the microbial consortium includes: 1) an engineered methanogen that contains a heterologous nucleic acid sequence encoding methyl-coenzyme M reductase derived from an anaerobic methane oxidizer, 2) an exoelectrogen microbe that produces electrically-conductive appendages and/or one or more types of electron carrier, and 3) a sludge, methane-acclimated sludge, a sludge isolate component, a methane-acclimated sludge isolate component chosen from Paracoccus spp., Geotoga spp., Geobacter spp., Methanosarcina spp., Garciella spp., humic acids; or a combination of any two or more thereof.

Wood, Thomas K.↗

Biomethanation to Upgrade Biogas to Pipeline Grade Methane

NREL is working closely with DOE, Electrochaea GmbH, and Southern California Gas Company (SoCalGas) to reduce costs of a biomethanation process capable of megawatt-scale deployment that upgrades organic biogas waste streams to produce pipeline quality renewable natural gas (RNG). Biomethanation is a two-step process using a single-celled methanogenic archaea that converts low-carbon low-cost hydrogen (H2) and waste carbon dioxide (CO2) to produce renewable methane (CH4). The process upgrades the biogenic CO2 - while allowing the CH4 to pass through - from biogas sources like dairies, wastewater treatment plants, and landfills. The CH4 produced is a drop-in direct replacement fuel and producers can participate in the growing number of carbon markets; like California's Low Carbon Fuel Standard and the Federal Renewable Fuel Standard. NREL and Argonne National Laboratory have completed a life cycle analysis using the GREET model to show that the biomethanation process produces RNG that is carbon negative even when H2 production via low-temperature water electrolysis is driven by the existing carbon intensity of California's electricity grid. And of course, even further carbon negative (-233 kg CO2e/kWh) when the electricity is produced from low-carbon sources like wind and solar. Leveraging lessons learned from operating SoCalGas' 700L 18-bar bioreactor system, NREL is designing and building a flexible RD&D platform that will enable field trials at biogas and other CO2 sources. A custom 16' long trailer will house a 20L 18-bar bioreactor, 3 - 25 kW proton exchange membrane electrolyzer, and dosing, thermal, and controls systems to support operations with only power, biogas, and water feedstocks required by the field locations. The end-of-project goal is to demonstrate pipeline quality RNG production (> 95% CH4, < 4% H2, <1% CO2, < 0.2% O2 and < 4 parts per million H2 sulfide) using real biogas feedstocks - thereby recycling both greenhouse gases for injection into the natural gas network or to be used onsite.

biogas upgrading↗

Synthesis and oxidation of methane

The present disclosure describes genes and proteins of the coenzyme F430 synthetic pathway. The genes and proteins in the pathway find uses as isolated nucleic acids, transformation vectors, a transformation media, genetically modified cells, methods of modulating methanogenesis, methods of modulating methane oxidation, methods of making a tetrapyrrole compound, methods of oxidizing methane, methods of biogenic methane synthesis is provided, methods of assaying an organism for potential methanogenic or methanotrophic activity, and isolated proteins.

Mansoorabadi, Steven↗

Biological upgrading of biogas assisted with membrane supplied hydrogen gas in a three-phase upflow reactor

Biogas upgrading via CO 2 conversion to CH 4 is an emerging technology for renewable natural gas production and carbon management, but its development is limited by the low H 2 gas to liquid phase transfer. Herein, an innovative biogas upgrading system employing a three-phase design was studied for CO 2 conversion with H 2 supply via gas-permeable membrane. The system produced biogas consisted of 74.1 ± 7.1 % CH 4 and 25.9 ± 7.1 % CO 2 with intermittent injection of H 2 . When H 2 supply was continuous, the CH 4 content increased to 91.6 ± 2.2 % at a H 2 :CO 2 ratio of 4.4. Although a higher ratio of 5.5 could result in a higher CH 4 percentage of 95.2 ± 2.5 %, biogas production rate started to decrease. The removal efficiency of organic contents remained above 90 % throughout the experiment. Microbial community analysis corroborated the findings, showing that hydrogenotrophic Methanobacteriaceae was more prevalent in the biofilm (71.9 %) compared to that in anaerobic digestion (15.8 %) and effluent (14.1 %).

Agriculture↗

Methane flux from transplanted soil monoliths depends on moisture, but not origin

Soils both produce and consume methane (CH 4 ), a potent greenhouse gas that contributes to climate change. In coastal forests, upland soils are shifting from being CH 4 sinks to sources as sea levels rise, increasingly flooding soils with little prior inundation history. Ecosystem CH 4 budgets are highly uncertain due in part to the difficulty in separating fluxes measured at the soil surface into individual production and consumption processes which are likely to have different responses to future environmental conditions. Here, we measured growing season CH 4 fluxes from soil monoliths transplanted four years prior along an inundation and salinity gradient to determine how changes in abiotic conditions control CH 4 flux rates. To parse net fluxes measured at the soil surface into their component gross rates, we paired field measurements with a stable isotope pool dilution incubation of surface soils. Throughout the growing season, net soil surface CH 4 flux was positively correlated with soil moisture (p < 0.01), with lowland-located soils tending towards CH 4 sources (mean 0.349 ± 1.11 mg CH 4 -C m -2 hr -1 , error is standard deviation) and upland-located soils tending towards CH 4 sinks (-0.003 ± 0.003 mg CH 4 -C m -2 hr -1 ). Transplanted soils’ fluxes were statistically identical to their native neighbors once microtopography-driven differences in soil moisture were controlled for. The pool dilution experiment revealed that production and consumption rates were similar in upland and lowland surface soils (2.82 ± 3.29 µmol CH 4 g -1 dry soil d -1 production, 3.47 ± 2.03 µmol CH 4 g -1 dry soil d -1 consumption), indicating the majority of production likely occurs at depth in lowland soils. Both gross and net fluxes from transplanted soils showed no effect of soil origin after four years, suggesting low resistance of CH 4 cycling to global change drivers. Our results indicate the strength of the coastal forest CH 4 sink is likely to decrease in proportion to sea-level rise.

59 BASIC BIOLOGICAL SCIENCES↗