Quantification of methanogenic heterodisulfide reductase activity in biogas sludge

Refai, Sarah; Berger, Stefanie; Wassmann, Kati; Deppenmeier, Uwe

doi:10.1016/j.jbiotec.2014.04.001

Cited by 8 publications

(4 citation statements)

References 15 publications

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“…For the aerobic determination of specific enzyme activities of Ack, Buk and But, as well as Western blot analysis of Ack, samples stored at −20°C were thawed on ice by the addition of one volume of corresponding enzyme assay reaction buffer (see below). Homogenization of the cells was carried out by using an ultrasonic pulser (Sonifier 250, Branson, Dietzenbach, Germany) with the following adjustments: 30 s pulsing at 60–70% intensity, 30 s break, 15 cycles (modified; Refai et al ., 2014 ). Cell debris was pelleted by centrifugation (4000 × g , 20 min, 4°C).…”

Section: Methodsmentioning

confidence: 99%

Analysis of the key enzymes of butyric and acetic acid fermentation in biogas reactors

Gabris

Bengelsdorf

Dürre

2015

Microbial Biotechnology

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This study aimed at the investigation of the mechanisms of acidogenesis, which is a key process during anaerobic digestion. To expose possible bottlenecks, specific activities of the key enzymes of acidification, such as acetate kinase (Ack, 0.23–0.99 U mg−1 protein), butyrate kinase (Buk, < 0.03 U mg−1 protein) and butyryl-CoA:acetate-CoA transferase (But, 3.24–7.64 U mg−1 protein), were determined in cell free extracts of biogas reactor content from three different biogas reactors. Furthermore, the detection of Ack was successful via Western blot analysis. Quantification of corresponding functional genes encoding Buk (buk) and But (but) was not feasible, although an amplification was possible. Thus, phylogenetic trees were constructed based on respective gene fragments. Four new clades of possible butyrate-producing bacteria were postulated, as well as bacteria of the genera Roseburia or Clostridium identified. The low Buk activity was in contrast to the high specific But activity in the analysed samples. Butyrate formation via Buk activity does barely occur in the investigated biogas reactor. Specific enzyme activities (Ack, Buk and But) in samples drawn from three different biogas reactors correlated with ammonia and ammonium concentrations (NH3 and NH4+-N), and a negative dependency can be postulated. Thus, high concentrations of NH3 and NH4+-N may lead to a bottleneck in acidogenesis due to decreased specific acidogenic enzyme activities.

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Section: Methodsmentioning

confidence: 99%

Analysis of the key enzymes of butyric and acetic acid fermentation in biogas reactors

Gabris

Bengelsdorf

Dürre

2015

Microbial Biotechnology

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“…In the reaction in which anaerobic oxidation of methane (AOM) directly generates CH 4 , methyl-coenzyme M-reductase (Mcr) uses coenzyme B (HS-CoB), which is an electron donor, and reduces the terminal methyl carrier CH 3 -S-CoM to produce methane ( Figure 6, Reaction (12)). In Reaction (13), heterodisulfide reductase (Hdr), as a key enzyme, can re-release coenzyme M and coenzyme B so that the methanogenesis process can be carried out continuously [87,89]. There are two types of Hdr [89,90].…”

Section: The Utilization Of Fe 2+ In Methanogans and Srbmentioning

confidence: 99%

“…In Reaction (13), heterodisulfide reductase (Hdr), as a key enzyme, can re-release coenzyme M and coenzyme B so that the methanogenesis process can be carried out continuously [87,89]. There are two types of Hdr [89,90]. One is necessary for methanogens using the reduced ferredoxin as electron donors.…”

Section: The Utilization Of Fe 2+ In Methanogans and Srbmentioning

confidence: 99%

Can Primary Ferroan Dolomite and Ankerite Be Precipitated? Its Implications for Formation of Submarine Methane-Derived Authigenic Carbonate (MDAC) Chimney

You

et al. 2019

Minerals

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Microbes can mediate the precipitation of primary dolomite under surface conditions. Meanwhile, primary dolomite mediated by microbes often contains more Fe2+ than standard dolomite in modern microbial culture experiments. Ferroan dolomite and ankerite have been regarded as secondary products. This paper reviews the process and possible mechanisms of microbial mediated precipitation of primary ferroan dolomite and/or ankerite. In the microbial geochemical Fe cycle, many dissimilatory iron-reducing bacteria (DIRB), sulfate-reducing bacteria (SRB), and methanogens can reduce Fe3+ to Fe2+, while SRB and methanogens can also promote the precipitation of primary dolomite. There are an oxygen respiration zone (ORZ), an iron reduction zone (IRZ), a sulfate reduction zone (SRZ), and a methanogenesis zone (MZ) from top to bottom in the muddy sediment diagenesis zone. DIRB in IRZ provide the lower section with Fe2+, which composes many enzymes and proteins to participate in metabolic processes of SRB and methanogens. Lastly, heterogeneous nucleation of ferroan dolomite on extracellular polymeric substances (EPS) and cell surfaces is mediated by SRB and methanogens. Exploring the origin of microbial ferroan dolomite may help to solve the “dolomite problem”.

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“…Therefore, enzyme activity assays, e.g. enzymes of hydrolysis (Gasch et al ., ) or enzymes of methanogenesis (Refai et al ., ), were also established for analysis of microbial communities from BGPs and could be applied to confirm metaproteome data. Alternatively, concentrations of intracellular and extracellular metabolites (metabolome) could be determined as they also represent the microbial activity.…”

Section: Tools For the Characterization Of Microbial Communitiesmentioning

confidence: 99%

Metaproteomics of complex microbial communities in biogas plants

Heyer

Kohrs

Reichl

et al. 2015

Microbial Biotechnology

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Production of biogas from agricultural biomass or organic wastes is an important source of renewable energy. Although thousands of biogas plants (BGPs) are operating in Germany, there is still a significant potential to improve yields, e.g. from fibrous substrates. In addition, process stability should be optimized. Besides evaluating technical measures, improving our understanding of microbial communities involved into the biogas process is considered as key issue to achieve both goals. Microscopic and genetic approaches to analyse community composition provide valuable experimental data, but fail to detect presence of enzymes and overall metabolic activity of microbial communities. Therefore, metaproteomics can significantly contribute to elucidate critical steps in the conversion of biomass to methane as it delivers combined functional and phylogenetic data. Although metaproteomics analyses are challenged by sample impurities, sample complexity and redundant protein identification, and are still limited by the availability of genome sequences, recent studies have shown promising results. In the following, the workflow and potential pitfalls for metaproteomics of samples from full-scale BGP are discussed. In addition, the value of metaproteomics to contribute to the further advancement of microbial ecology is evaluated. Finally, synergistic effects expected when metaproteomics is combined with advanced imaging techniques, metagenomics, metatranscriptomics and metabolomics are addressed.

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Quantification of methanogenic heterodisulfide reductase activity in biogas sludge

Cited by 8 publications

References 15 publications

Analysis of the key enzymes of butyric and acetic acid fermentation in biogas reactors

Analysis of the key enzymes of butyric and acetic acid fermentation in biogas reactors

Can Primary Ferroan Dolomite and Ankerite Be Precipitated? Its Implications for Formation of Submarine Methane-Derived Authigenic Carbonate (MDAC) Chimney

Metaproteomics of complex microbial communities in biogas plants

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