Showing papers on "Methanogen published in 2017"

Rumen methanogens and mitigation of methane emission by anti-methanogenic compounds and substances

Abstract: Methanogenic archaea reside primarily in the rumen and the lower segments of the intestines of ruminants, where they utilize the reducing equivalents derived from rumen fermentation to reduce carbon dioxide, formic acid, or methylamines to methane (CH4). Research on methanogens in the rumen has attracted great interest in the last decade because CH4 emission from ruminants contributes to global greenhouse gas emission and represents a loss of feed energy. Some DNA-based phylogenetic studies have depicted a diverse and dynamic community of methanogens in the rumen. In the past decade, researchers have focused on elucidating the underpinning that determines and affects the diversity, composition, structure, and dynamics of methanogen community of the rumen. Concurrently, many researchers have attempted to develop and evaluate interventions to mitigate enteric CH4 emission. Although much work has been done using plant secondary metabolites, other approaches such as using nitrate and 3-nitrooxy propanol have also yielded promising results. Most of these antimethanogenic compounds or substances often show inconsistent results among studies and also lead to adverse effects on feed intake and digestion and other aspects of rumen fermentation when fed at doses high enough to achieve effective mitigation. This review provides a brief overview of the rumen methanogens and then an appraisal of most of the antimethanogenic compounds and substances that have been evaluated both in vitro and in vivo. Knowledge gaps and future research needs are also discussed with a focus on methanogens and methane mitigation.

Methanogenesis in oxygenated soils is a substantial fraction of wetland methane emissions.

Abstract: The current paradigm, widely incorporated in soil biogeochemical models, is that microbial methanogenesis can only occur in anoxic habitats. In contrast, here we show clear geochemical and biological evidence for methane production in well-oxygenated soils of a freshwater wetland. A comparison of oxic to anoxic soils reveal up to ten times greater methane production and nine times more methanogenesis activity in oxygenated soils. Metagenomic and metatranscriptomic sequencing recover the first near-complete genomes for a novel methanogen species, and show acetoclastic production from this organism was the dominant methanogenesis pathway in oxygenated soils. This organism, Candidatus Methanothrix paradoxum, is prevalent across methane emitting ecosystems, suggesting a global significance. Moreover, in this wetland, we estimate that up to 80% of methane fluxes could be attributed to methanogenesis in oxygenated soils. Together, our findings challenge a widely held assumption about methanogenesis, with significant ramifications for global methane estimates and Earth system modeling.

Energy Metabolism during Anaerobic Methane Oxidation in ANME Archaea.

Abstract: Anaerobic methane oxidation in archaea is often presented to operate via a pathway of "reverse methanogenesis". However, if the cumulative reactions of a methanogen are run in reverse there is no apparent way to conserve energy. Recent findings suggest that chemiosmotic coupling enzymes known from their use in methylotrophic and acetoclastic methanogens-in addition to unique terminal reductases-biochemically facilitate energy conservation during complete CH4 oxidation to CO2. The apparent enzyme modularity of these organisms highlights how microbes can arrange their energy metabolisms to accommodate diverse chemical potentials in various ecological niches, even in the extreme case of utilizing "reverse" thermodynamic potentials.

A Ferredoxin- and F420H2-Dependent, Electron-Bifurcating, Heterodisulfide Reductase with Homologs in the Domains Bacteria and Archaea.

Abstract: Heterodisulfide reductases (Hdr) of the HdrABC class are ancient enzymes and a component of the anaerobic core belonging to the prokaryotic common ancestor. The ancient origin is consistent with the widespread occurrence of genes encoding putative HdrABC homologs in metabolically diverse prokaryotes predicting diverse physiological functions; however, only one HdrABC has been characterized and that was from a narrow metabolic group of obligate CO2-reducing methanogenic anaerobes (methanogens) from the domain Archaea Here we report the biochemical characterization of an HdrABC homolog (HdrA2B2C2) from the acetate-utilizing methanogen Methanosarcina acetivorans with unusual properties structurally and functionally distinct from the only other HdrABC characterized. Homologs of the HdrA2B2C2 archetype are present in phylogenetically and metabolically diverse species from the domains Bacteria and Archaea The expression of the individual HdrA2, HdrB2, and HdrB2C2 enzymes in Escherichia coli, and reconstitution of an active HdrA2B2C2 complex, revealed an intersubunit electron transport pathway dependent on ferredoxin or coenzyme F420 (F420H2) as an electron donor. Remarkably, HdrA2B2C2 couples the previously unknown endergonic oxidation of F420H2 and reduction of ferredoxin with the exergonic oxidation of F420H2 and reduction of the heterodisulfide of coenzyme M and coenzyme B (CoMS-SCoB). The unique electron bifurcation predicts a role for HdrA2B2C2 in Fe(III)-dependent anaerobic methane oxidation (ANME) by M. acetivorans and uncultured species from ANME environments. HdrA2B2C2, ubiquitous in acetotrophic methanogens, was shown to participate in electron transfer during acetotrophic growth of M. acetivorans and proposed to be essential for growth in the environment when acetate is limiting. IMPORTANCE Discovery of the archetype HdrA2B2C2 heterodisulfide reductase with categorically unique properties extends the understanding of this ancient family beyond CO2-reducing methanogens to include diverse prokaryotes from the domains Bacteria and Archaea The unprecedented coenzyme F420-dependent electron bifurcation, an emerging fundamental principle of energy conservation, predicts a role for HdrA2B2C2 in diverse metabolisms, including anaerobic CH4-oxidizing pathways. The results document an electron transport role for HdrA2B2C2 in acetate-utilizing methanogens responsible for at least two-thirds of the methane produced in Earth's biosphere. The previously unavailable heterologous production of individual subunits and the reconstitution of HdrA2B2C2 with activity have provided an understanding of intersubunit electron transfer in the HdrABC class and a platform for investigating the principles of electron bifurcation.

Qualitative Analysis of Microbial Dynamics during Anaerobic Digestion of Microalgal Biomass in a UASB Reactor.

Abstract: Anaerobic digestion (AD) is a microbiologically coordinated process with dynamic relationships between bacterial players. Current understanding of dynamic changes in the bacterial composition during the AD process is incomplete. The objective of this research was to assess changes in bacterial community composition that coordinates with anaerobic codigestion of microalgal biomass cultivated on municipal wastewater. An upflow anaerobic sludge blanket reactor was used to achieve high rates of microalgae decomposition and biogas production. Samples of the sludge were collected throughout AD and extracted DNA was subjected to next-generation sequencing using methanogen mcrA gene specific and universal bacterial primers. Analysis of the data revealed that samples taken at different stages of AD had varying bacterial composition. A group consisting of Bacteroidales, Pseudomonadales, and Enterobacteriales was identified to be putatively responsible for the hydrolysis of microalgal biomass. The methanogenesis phase was dominated by Methanosarcina mazei. Results of observed changes in the composition of microbial communities during AD can be used as a road map to stimulate key bacterial species identified at each phase of AD to increase yield of biogas and rate of substrate decomposition. This research demonstrates a successful exploitation of methane production from microalgae without any biomass pretreatment.

Microbial methanogenesis in the sulfate-reducing zone of sediments in the Eckernförde Bay, SW Baltic Sea

Abstract: . Benthic microbial methanogenesis is a known source of methane in marine systems. In most sediments, the majority of methanogenesis is located below the sulfate-reducing zone, as sulfate reducers outcompete methanogens for the major substrates hydrogen and acetate. The coexistence of methanogenesis and sulfate reduction has been shown before and is possible through the usage of noncompetitive substrates by methanogens such as methanol or methylated amines. However, knowledge about the magnitude, seasonality, and environmental controls of this noncompetitive methane production is sparse. In the present study, the presence of methanogenesis within the sulfate reduction zone (SRZ methanogenesis) was investigated in sediments (0–30 cm below seafloor, cm b.s.f.) of the seasonally hypoxic Eckernforde Bay in the southwestern Baltic Sea. Water column parameters such as oxygen, temperature, and salinity together with porewater geochemistry and benthic methanogenesis rates were determined in the sampling area Boknis Eck quarterly from March 2013 to September 2014 to investigate the effect of seasonal environmental changes on the rate and distribution of SRZ methanogenesis, to estimate its potential contribution to benthic methane emissions, and to identify the potential methanogenic groups responsible for SRZ methanogenesis. The metabolic pathway of methanogenesis in the presence or absence of sulfate reducers, which after the addition of a noncompetitive substrate was studied in four experimental setups: (1) unaltered sediment batch incubations (net methanogenesis), (2) 14C-bicarbonate labeling experiments (hydrogenotrophic methanogenesis), (3) manipulated experiments with the addition of either molybdate (sulfate reducer inhibitor), 2-bromoethanesulfonate (methanogen inhibitor), or methanol (noncompetitive substrate, potential methanogenesis), and (4) the addition of 13C-labeled methanol (potential methylotrophic methanogenesis). After incubation with methanol, molecular analyses were conducted to identify key functional methanogenic groups during methylotrophic methanogenesis. To also compare the magnitudes of SRZ methanogenesis with methanogenesis below the sulfate reduction zone (> 30 cm b.s.f.), hydrogenotrophic methanogenesis was determined by 14C-bicarbonate radiotracer incubation in samples collected in September 2013. SRZ methanogenesis changed seasonally in the upper 30 cm b.s.f. with rates increasing from March (0.2 nmol cm−3 d−1) to November (1.3 nmol cm−3 d−1) 2013 and March (0.2 nmol cm−3 d−1) to September (0.4 nmol cm−3 d−1) 2014. Its magnitude and distribution appeared to be controlled by organic matter availability, C / N, temperature, and oxygen in the water column, revealing higher rates in the warm, stratified, hypoxic seasons (September–November) compared to the colder, oxygenated seasons (March–June) of each year. The majority of SRZ methanogenesis was likely driven by the usage of noncompetitive substrates (e.g., methanol and methylated compounds) to avoid competition with sulfate reducers, as was indicated by the 1000–3000-fold increase in potential methanogenesis activity observed after methanol addition. Accordingly, competitive hydrogenotrophic methanogenesis increased in the sediment only below the depth of sulfate penetration (> 30 cm b.s.f.). Members of the family Methanosarcinaceae, which are known for methylotrophic methanogenesis, were detected by PCR using Methanosarcinaceae-specific primers and are likely to be responsible for the observed SRZ methanogenesis. The present study indicates that SRZ methanogenesis is an important component of the benthic methane budget and carbon cycling in Eckernforde Bay. Although its contributions to methane emissions from the sediment into the water column are probably minor, SRZ methanogenesis could directly feed into methane oxidation above the sulfate–methane transition zone.

Methylphosphonate metabolism by Pseudomonas sp. populations contributes to the methane oversaturation paradox in an oxic freshwater lake.

Abstract: The 'CH4 oversaturation paradox' has been observed in oxygen-rich marine and lake waters, and viewed to significantly contribute to biosphere cycling of methane, a potent greenhouse gas. Our study focused on the intriguing well-defined pelagic methane enriched zone (PMEZ) in freshwater lakes. Spiking Yellowstone Lake PMEZ samples with 13 C-labeled potential methanogenesis substrates found only 13 C-methylphosphonate (MPn) resulted in 13 CH4 generation. In 16S rRNA gene Illumina libraries, four Pseudomonas sp. operational taxonomic units surprisingly accounted for ∼11% abundance in the PMEZ community. Pseudomonas sp. isolates were also obtained from MPn enrichments with PMEZ water; they were most aggressive in MPn metabolism and their 16S rRNA gene sequences matched 35% of the Illumina PMEZ Pseudomonas reads. Further, two key genes encoding C-P lyase (phnJL, an important enzyme for dealkylation of MPn), were only amplifiable from PMEZ DNA and all PCR generated phnJL clones matched those of the Pseudomonas sp. isolates. Notably, methanogen 16S rRNA signatures were absent in all Illumina libraries and mcrA was not detected via PCR. Collectively, these observations are consistent with the conclusion that MPn metabolism contributes significantly to CH4 oversaturation in Yellowstone Lake and likely other oxic freshwater lake environments, and that Pseudomonas sp. populations are critical participants.

Enhanced system performance by dosing ferroferric oxide during the anaerobic treatment of tryptone-based high-strength wastewater.

Abstract: Conductive materials can facilitate syntrophic methane (CH4) production by improving direct interspecies electron transfer. The effect of a conductive material, ferroferric oxide (Fe3O4), on the anaerobic treatment of tryptone-based high-strength wastewater was investigated in two anaerobic sequencing batch reactors (ASBRs). One was the control reactor without amendment of Fe3O4, and the other was the Fe3O4-amended reactor. In an ASBR reaction cycle, the dosage of Fe3O4 increased the CH4 production rate by 12.2%, shortened the methanogenic lag phase by 32.3%, and improved the COD removal rate by 8.1%. During anaerobic treatment, the dosage of Fe3O4 not only enhanced the CH4 production from acetate but also facilitated the hydrolysis/acidification process fed by tryptone. Maximum electron transport activities were increased by 40.4, 137.3, and 64.6% in the processes of the ASBR reaction cycle, the CH4 production from acetate, and the hydrolysis/acidification of tryptone, respectively. Among all tested quorum sensing signal substances, N-octanoyl-l-homoserine lactone was dominant in both ASBRs, with the highest concentration in the biomass and the lowest concentration in the water phase. In addition, the concentration of signal substance was high in the ASBR cycle and low in the CH4 production from acetate and the hydrolysis/acidification of tryptone. Proteiniclasticum and vadinCA02 were dominant acidogens, and their relative abundances increased during the long-term operation with Fe3O4 dosing. Methanosarcina was the predominant methanogen, contributing to interspecies electron transfer.

Microbial community changes in methanogenic granules during the transition from mesophilic to thermophilic conditions.

Abstract: Upflow anaerobic sludge blanket (UASB) reactor is one of the most applied technologies for various high-strength wastewater treatments. The present study analysed the microbial community changes in UASB granules during the transition from mesophilic to thermophilic conditions. Dynamicity of microbial community in granules was analysed using high-throughput sequencing of 16S ribosomal RNA gene amplicons, and the results showed that the temperature strictly determines the diversity of the microbial consortium. It was demonstrated that most of the microbes which were present in the initial mesophilic community were not found in the granules after the transition to thermophilic conditions. More specifically, only members from family Anaerolinaceae managed to tolerate the temperature change and contributed in maintaining the physical integrity of granular structure. On the contrary, new hydrolytic and fermentative bacteria were quickly replacing the old members in the community. A direct result from this abrupt change in the microbial diversity was the accumulation of volatile fatty acids and the concomitant pH drop in the reactor inhibiting the overall anaerobic digestion process. Nevertheless, by maintaining deliberately the pH levels at values higher than 6.5, a methanogen belonging to Methanoculleus genus emerged in the community enhancing the methane production.

Microbial community characterization during anaerobic digestion of Scenedesmus spp. under mesophilic and thermophilic conditions

Abstract: Microbial communities were thoroughly characterized in a mesophilic anaerobic membrane bioreactor (AnMBR) and a thermophilic continuous stirred tank reactor (CSTR), which were both treating recalcitrant microalgal biomass dominated by Scenedesmus . 16S rRNA amplicon sequencing analysis was performed when the AnMBR achieved 70% algal biodegradation and revealed high microbial diversity, probably due to the high solid retention time (SRT) of the AnMBR configuration. The bacterial community consisted of Chloroflexi (27.9%), WWE1 (19.0%) and Proteobacteria (15.4%) as the major phyla, followed by Spirochaetes (7.7%), Bacteroidetes (5.9%) and Firmicutes (3.6%). These phyla are known to exhibit proteolytic and cellulolytic capabilities required to degrade the Scenedesmus cell-wall. Methanosaeta was the most abundant methanogen detected in the AnMBR suggesting that methane was mainly produced by the acetoclastic pathway. In comparison, the thermophilic CSTR achieved 32.6% algal biodegradation, and its bacterial community had fewer Operational Taxonomic Units (977 OTUs) than the AnMBR (1396 OTUs), as is generally observed for high temperature biogas reactors. However, phyla with high hydrolytic potential were detected such as Firmicutes (34.6%) and the candidate taxon EM3 (38.7%) in the thermophilic CSTR. Although the functional metabolism of EM3 in anaerobic digesters is unknown, the high abundance of EM3 suggests that this taxon plays an important role in the thermophilic, anaerobic degradation of Scenedesmus . The abundant syntrophic bacteria and the detection of hydrogenotrophic methanogens in the thermophilic CSTR suggest that the hydrogenotrophic pathway was the dominant pathway for methane production in this reactor.

Investigation and manipulation of metabolically active methanogen community composition during rumen development in black goats.

Abstract: This study was performed to investigate the initial colonization of metabolically active methanogens and subsequent changes in four fractions: the rumen solid-phase (RS), liquid-phase (RL), protozoa-associated (RP), and epithelium-associated (RE) from 1 to 60 d after birth, and manipulate methanogen community by early weaning on 40 d and supplementing rhubarb from 40 to 60 d in black goats. The RNA-based real-time quantitative PCR and 16S rRNA amplicon sequencing were employed to indicate the metabolically active methanogens. Results showed that active methanogens colonized in RL and RE on 1 d after birth. RP and RE contained the highest and lowest density of methanogens, respectively. Methanobrevibacter, Candidatus Methanomethylophilus, and Methanosphaera were the top three genera. The methanogen communities before weaning differed from those post weaning and the structure of the methanogen community in RE was distinct from those in the other three fractions. The discrepancies in the distribution of methanogens across four fractions, and various fluctuations in abundances among four fractions according to age were observed. The addition of rhubarb significantly (P < 0.05) reduced the abundances of Methanimicrococcus spp. in four fractions on 50 d, but did not change the methanogen community composition on 60 d.

Non-autotrophic methanogens dominate in anaerobic digesters.

Abstract: Anaerobic digesters are man-made habitats for fermentative and methanogenic microbes, and are characterized by extremely high concentrations of organics. However, little is known about how microbes adapt to such habitats. In the present study, we report phylogenetic, metagenomic, and metatranscriptomic analyses of microbiomes in thermophilic packed-bed digesters fed acetate as the major substrate, and we have shown that acetoclastic and hydrogenotrophic methanogens that utilize acetate as a carbon source dominate there. Deep sequencing and precise binning of the metagenomes reconstructed complete genomes for two dominant methanogens affiliated with the genera Methanosarcina and Methanothermobacter, along with 37 draft genomes. The reconstructed Methanosarcina genome was almost identical to that of a thermophilic acetoclastic methanogen Methanosarcina thermophila TM-1, indicating its cosmopolitan distribution in thermophilic digesters. The reconstructed Methanothermobacter (designated as Met2) was closely related to Methanothermobacter tenebrarum, a non-autotrophic hydrogenotrophic methanogen that grows in the presence of acetate. Met2 lacks the Cdh complex required for CO2 fixation, suggesting that it requires organic molecules, such as acetate, as carbon sources. Although the metagenomic analysis also detected autotrophic methanogens, they were less than 1% in abundance of Met2. These results suggested that non-autotrophic methanogens preferentially grow in anaerobic digesters containing high concentrations of organics.

Genomic analysis of methanogenic archaea reveals a shift towards energy conservation

Abstract: The metabolism of archaeal methanogens drives methane release into the environment and is critical to understanding global carbon cycling. Methanogenesis operates at a very low reducing potential compared to other forms of respiration and is therefore critical to many anaerobic environments. Harnessing or altering methanogen metabolism has the potential to mitigate global warming and even be utilized for energy applications. Here, we report draft genome sequences for the isolated methanogens Methanobacterium bryantii, Methanosarcina spelaei, Methanosphaera cuniculi, and Methanocorpusculum parvum. These anaerobic, methane-producing archaea represent a diverse set of isolates, capable of methylotrophic, acetoclastic, and hydrogenotrophic methanogenesis. Assembly and analysis of the genomes allowed for simple and rapid reconstruction of metabolism in the four methanogens. Comparison of the distribution of Clusters of Orthologous Groups (COG) proteins to a sample of genomes from the RefSeq database revealed a trend towards energy conservation in genome composition of all methanogens sequenced. Further analysis of the predicted membrane proteins and transporters distinguished differing energy conservation methods utilized during methanogenesis, such as chemiosmotic coupling in Msar. spelaei and electron bifurcation linked to chemiosmotic coupling in Mbac. bryantii and Msph. cuniculi. Methanogens occupy a unique ecological niche, acting as the terminal electron acceptors in anaerobic environments, and their genomes display a significant shift towards energy conservation. The genome-enabled reconstructed metabolisms reported here have significance to diverse anaerobic communities and have led to proposed substrate utilization not previously reported in isolation, such as formate and methanol metabolism in Mbac. bryantii and CO2 metabolism in Msph. cuniculi. The newly proposed substrates establish an important foundation with which to decipher how methanogens behave in native communities, as CO2 and formate are common electron carriers in microbial communities.

Metabolic engineering of Methanosarcina acetivorans for lactate production from methane.

Abstract: We previously demonstrated anaerobic conversion of the greenhouse gas methane into acetate using an engineered archaeon that produces methyl-coenzyme M reductase (Mcr) from unculturable microorganisms from a microbial mat in the Black Sea to create the first culturable prokaryote that reverses methanogenesis and grows anaerobically on methane. In this work, we further engineered the same host with the goal of converting methane into butanol. Instead, we discovered a process for converting methane to a secreted valuable product, L-lactate, with sufficient optical purity for synthesizing the biodegradable plastic poly-lactic acid. We determined that the 3-hydroxybutyryl-CoA dehydrogenase (Hbd) from Clostridium acetobutylicum is responsible for lactate production. This work demonstrates the first metabolic engineering of a methanogen with a synthetic pathway; in effect, we produce a novel product (lactate) from a novel substrate (methane) by cloning the three genes for Mcr and one for Hbd. We further demonstrate the utility of anaerobic methane conversion with an increased lactate yield compared to aerobic methane conversion to lactate. Biotechnol. Bioeng. 2017;114: 852–861. © 2016 Wiley Periodicals, Inc.

Unexpected competitiveness of Methanosaeta populations at elevated acetate concentrations in methanogenic treatment of animal wastewater

Abstract: Acetoclastic methanogenesis is a key metabolic process in anaerobic digestion, a technology with broad applications in biogas production and waste treatment. Acetoclastic methanogenesis is known to be performed by two archaeal genera, Methanosaeta and Methanosarcina. The conventional model posits that Methanosaeta populations are more competitive at low acetate levels (<1 mM) than Methanosarcina and vice versa at higher acetate concentrations. While this model is supported by an extensive body of studies, reports of inconsistency have grown that Methanosaeta were observed to outnumber Methanosarcina at elevated acetate levels. In this study, monitoring of anaerobic digesters treating animal wastewater unexpectedly identified Methanosaeta as the dominant acetoclastic methanogen population at both low and high acetate levels during organic overloading. The surprising competitiveness of Methanosaeta at elevated acetate was further supported by the enrichment of Methanosaeta with high concentrations of acetate (20 mM). The dominance of Methanosaeta in the methanogen community could be reproduced in anaerobic digesters with the direct addition of acetate to above 20 mM, again supporting the competitiveness of Methanosaeta over Methanosarcina at elevated acetate levels. This study for the first time systematically demonstrated that the dominance of Methanosaeta populations in anaerobic digestion could be linked to the competitiveness of Methanosaeta at elevated acetate concentrations. Given the importance of acetoclastic methanogenesis in biological methane production, findings from this study could have major implications for developing strategies for more effective control of methanogenic treatment processes.

Nitrate decreases methane production also by increasing methane oxidation through stimulating NC10 population in ruminal culture

Abstract: Studies proved that addition of nitrate in rumen could lead to reduction of methane emission. The mechanism of this function was involved in the competition effect of nitrate on hydrogen consumption and the inhibitory effect of generated nitrite on methanogen proliferation. The present study investigated an alternative mechanism that denitrifying anaerobic methane oxidizing (DAMO) bacteria, DAMO archaea and anammox bacteria may co-exist in rumen, therefore, more methane can be oxidized when addition of nitrate. Ruminal batch culture model was used to test the effects of addition of 5 mM NaNO3, 4 mM NH4Cl, or both into the culture substrate on methane production, fermentation patterns, and population of methanogens, NC10 and anaerobic methanotrophic-2d (ANME-2d). Our results showed that NC10 in the ruminal culture was detected by polymerase chain reaction (PCR) when using NC10 special primer sets, and addition of nitrate reduced methane production and the relative proportions of methanogen, whereas increased the relative proportion of NC10. A combined addition of ammonia salt and nitrate did not show further inhibitory effect on methane production but accelerated nitrate removal. We did not detect DAMO archaea in ruminal culture by real-time PCR when using ANME-2d special primer sets. The present study may encourage researchers to pay more attention to methane oxidation performed by anaerobic methanotroph when studying the strategies of inhibiting ruminal methane emission.

Indigenously associated methanogens intensified the metabolism in hydrogenosomes of anaerobic fungi with xylose as substrate.

Abstract: Anaerobic fungi are potent lignocellulose degraders, but have not yet been exploited in this capacity, largely owing to their poor metabolic characterization. In the current study, a time course of fermentation was conducted to study the effect of the co-cultured methanogens on xylose metabolism by anaerobic fungi. The fermentation end-products from anaerobic fungal monoculture were H2 (6.7 ml), CO2 (65.7 ml), formate (17.90 mM), acetate (9.00 mM), lactate (11.89 mM), ethanol, and malate after 96 h fermentation. Compared to the monoculture, the end-products of co-culture shifted to more CO2 (71.8 ml) and acetate (15.20 mM), methane (14.9 ml), less lactate (5.28 mM), and hardly detectable formate and H2 at the end of fermentation. After 48 h, accumulated formate was remarkably consumed by co-cultured methanogens, accompanied by significantly increased acetate, CO2 and pH, and decreased lactate and malate. Xylose utilization, in both cultures, was similar during fermentation. However, the relative flux of carbon in hydrogenosomes in the co-culture was higher than that in the monoculture. In conclusion, the co-culture with methanogens enhanced “energy yields” of anaerobic fungi by removing the accumulated formate, decreased the metabolism in cytosol, for example, the lactate pathway, and increased the metabolism in hydrogenosomes, for example, the acetate pathway.

Effects of ammonia on propionate degradation and microbial community in digesters using propionate as a sole carbon source

Abstract: BACKGROUND Propionate accumulation may lead to digester failure. This study aimed to investigate the effect of ammonia, a metabolic product of protein, on propionate degradation. The shift of microbial community was also investigated. RESULTS Propionate accumulated over the experimental period in the reactor with a total ammonia nitrogen (TAN) concentration of 2.5 g N L−1, as a result the digester failed after 4 hydraulic retention times (HRT) at an organic load rate (OLR) of 0.8 g propionic acid (HPr) L−1 d−1. The average HPr degradation rate was below 54% during the fourth HRT, while >97% of the degraded HPr was converted to methane. The reactor without ammonia stress did not experience HPr accumulation and OLR was increased stepwise to 1.2 g L−1 d−1 at the 8th HRT. The average HPr degradation rate and methane recovery rate of this reactor in the last HRT was 99% and 74%, respectively. According to the shifts of microbial community, acetoclastic methanogen was more vulnerable to ammonia than hydrogenotrophic methanogen CONCLUSION TAN concentration of 2.5 g N L−1 inhibited propionate degradation more severely than methanogenesis. The loss of abundance of Clostridiaceae and Syntrophobacter might be the main reason for the poor performance under ammonia stress. © 2017 Society of Chemical Industry

Hydrogen and volatile fatty acids production from marine macroalgae by anaerobic fermentation

Abstract: Hydrogen and volatile fatty acids (VFAs) were coproduced from marine macroalgae by anaerobic fermentation using a microbial community. The hydrogen and VFAs production were characterized based on inoculum heat-treatment, methanogen inhibitor addition, operating temperature, and in-situ extraction of VFAs. Maximum hydrogen of 179 mL/g-VS and VFAs concentration of 9.8 g/L were produced from 35 g/L of S. japonica within 5 days of anaerobic fermentation. Hydrogen and VFAs yields were well-correlated with carbohydrate content of substrate. Inoculum heat-treatment significantly improved hydrogen production while the VFAs productivity was affected adversely. The addition of methanogen inhibitors also enhanced the hydrogen production, but the effect on VFAs production was dependent on the type of inhibitor used. Low temperature (25°C) was found to be favorable for high hydrogen and VFAs yield, while high temperature (40°C) and programmed-temperature (35 ~ 45°C) were more favorable for hydrogen and VFAs productivity. Clostridium sp. content was found to be the most abundant at 25°C. An extractive fermentation with anion-exchange resin was tested to recover the VFAs and to control the pH during the anaerobic fermentation.