Energy Conservation Associated with Ethanol Formation from H 2 and CO 2 in Clostridium autoethanogenum Involving Electron Bifurcation
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UNLABELLED: Most acetogens can reduce CO2 with H2 to acetic acid via the Wood-Ljungdahl pathway, in which the ATP required for formate activation is regenerated in the acetate kinase reaction. However, a few acetogens, such as Clostridium autoethanogenum, Clostridium ljungdahlii, and Clostridium ragsdalei, also form large amounts of ethanol from CO2 and H2. How these anaerobes with a growth pH optimum near 5 conserve energy has remained elusive. We investigated this question by determining the specific activities and cofactor specificities of all relevant oxidoreductases in cell extracts of H2/CO2-grown C. autoethanogenum. The activity studies were backed up by transcriptional and mutational analyses. Most notably, despite the presence of six hydrogenase systems of various types encoded in the genome, the cells appear to contain only one active hydrogenase. The active [FeFe]-hydrogenase is electron bifurcating, with ferredoxin and NADP as the two electron acceptors. Consistently, most of the other active oxidoreductases rely on either reduced ferredoxin and/or NADPH as the electron donor. An exception is ethanol dehydrogenase, which was found to be NAD specific. Methylenetetrahydrofolate reductase activity could only be demonstrated with artificial electron donors. Key to the understanding of this energy metabolism is the presence of membrane-associated reduced ferredoxin:NAD(+) oxidoreductase (Rnf), of electron-bifurcating and ferredoxin-dependent transhydrogenase (Nfn), and of acetaldehyde:ferredoxin oxidoreductase, which is present with very high specific activities in H2/CO2-grown cells. Based on these findings and on thermodynamic considerations, we propose metabolic schemes that allow, depending on the H2 partial pressure, the chemiosmotic synthesis of 0.14 to 1.5 mol ATP per mol ethanol synthesized from CO2 and H2. IMPORTANCE: Ethanol formation from syngas (H2, CO, and CO2) and from H2 and CO2 that is catalyzed by bacteria is presently a much-discussed process for sustainable production of biofuels. Although the process is already in use, its biochemistry is only incompletely understood. The most pertinent question is how the bacteria conserve energy for growth during ethanol formation from H2 and CO2, considering that acetyl coenzyme A (acetyl-CoA), is an intermediate. Can reduction of the activated acetic acid to ethanol with H2 be coupled with the phosphorylation of ADP? Evidence is presented that this is indeed possible, via both substrate-level phosphorylation and electron transport phosphorylation. In the case of substrate-level phosphorylation, acetyl-CoA reduction to ethanol proceeds via free acetic acid involving acetaldehyde:ferredoxin oxidoreductase (carboxylate reductase).
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Transcriptomics analyses have been performed for C. autoethanogenum and C. ljungdahlii.
“and transcriptomics analyses of C. autoethanogenum (Mock et al., 2015; Marcellin et al., 2016) and C. ljungdahlii (Nagarajan et al., 2013; Tan et al., 2013; Whitham et al., 2015).”
From Gas Fermentation—A Flexible Platform for Commercial Scale Production of Low-Carbon-Fuels and Chemicals from Waste and Renewable Feedstocks (2016) · cites this paper as Mock et al., 2015
Cofactors for key reactions have been identified for C. autoethanogenum, A. woodii, M. thermoacetica, and E. limosum.
“identification of cofactors for key reactions for C. autoethanogenum (Wang et al., 2013a; Mock et al., 2015), A. woodii (Schuchmann and Müller, 2012, 2014), M. thermoacetica (Huang et al., 2012; Mock et al., 2014), and E. limosum (Jeong et al., 2015);”
From Gas Fermentation—A Flexible Platform for Commercial Scale Production of Low-Carbon-Fuels and Chemicals from Waste and Renewable Feedstocks (2016) · cites this paper as Mock et al., 2015
Conjugation was reported for C. autoethanogenum using E. coli strain HB101 as donor and for A. woodii using E. coli strain S17-1 as donor.
“Conjugation was reported for C. autoethanogenum (using E. coli strain HB101 as donor) (Mock et al., 2015), and A. woodii (using E. coli strain S17-1 as donor) (Strätz et al., 1994).”
From Gas Fermentation—A Flexible Platform for Commercial Scale Production of Low-Carbon-Fuels and Chemicals from Waste and Renewable Feedstocks (2016) · cites this paper as Mock et al., 2015
H2/CO2 gas fermentations mostly produce acetate, though production of ethanol and other products such as acetone or butyrate have been described, albeit at lower productivity compared to CO.
“H2/CO2 gas fermentations mostly produce acetate, though production of ethanol (Mock et al., 2015) and other products such as acetone (Schiel-Bengelsdorf and Dürre, 2012; Hoffmeister et al., 2016) or butyrate (Ueki et al., 2014) have been described, albeit at lower productivity compared to CO.”
From Gas Fermentation—A Flexible Platform for Commercial Scale Production of Low-Carbon-Fuels and Chemicals from Waste and Renewable Feedstocks (2016) · cites this paper as Mock et al., 2015
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- Authors
- Mock, J., Zheng, Y., Mueller, A. P., Ly, S., Tran, L., Segovia, S., Nagaraju, S., Köpke, M., Dürre, P., & Thauer, R. K.
- Affiliations
- Max Planck Institute for Terrestrial Microbiology (DE)
- LanzaTech Inc., Skokie, Illinois, USA
- Universität Ulm (DE)
- Venue
- Journal of Bacteriology
- Publisher
- American Society for Microbiology
- Published
- 2015-09-15
- Volume
- 197
- Issue
- 18
- Pages
- 2965-2980
- ISSN
- 0021-9193 (ISSN-L)
- 1098-5530
- PMC
- PMC4542177
- OpenAlex
- W1841900872
- Cited by
- 220
- References
- 88
- Citations by year
- 2026: 13 · 2025: 17 · 2024: 18 · 2023: 25 · 2022: 26 · 2021: 18 · 2020: 27 · 2019: 21 · 2018: 20 · 2017: 13 · 2016: 16 · 2015: 6
- FWCI
- 7.96
- Type
- journal article
- Language
- en
- Open access
- green
- www.ncbi.nlm.nih.gov
- Locations
- Journal of Bacteriology (publishedVersion)
- PubMed Central (publishedVersion)
- PubMed (publishedVersion)
- Max Planck Institute for Plasma Physics (submittedVersion)
- MPG.PuRe (Max Planck Society) (submittedVersion)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Microbial Metabolic Engineering and Bioproduction
- Physical Sciences › Engineering › Building and Construction › Anaerobic Digestion and Biogas Production
- Physical Sciences › Engineering › Biomedical Engineering › Biofuel production and bioconversion
- Keywords
- Clostridium autoethanogenum, ethanol production, electron bifurcation, energy conservation, acetogenic bacteria, hydrogenase, [FeFe]-hydrogenase, methylenetetrahydrofolate reductase, ethanol dehydrogenase, Wood-Ljungdahl pathway, ATP production, syngas conversion, biofuel production
- MeSH terms
- Acetyl Coenzyme A, Adenosine Diphosphate, Ethanol, Bacterial Proteins, Carbon Dioxide, Clostridium, Electron Transport, Energy Metabolism, Hydrogen, Membrane Proteins, Oxidoreductases, Phosphoproteins, Gene Expression Regulation, Bacterial, Gene Expression Regulation, Enzymologic, Acetic Acid
About these checks
Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.
Statistics and arithmetic are recomputed, not judged.
Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.
What counts as a claim ›How we measure confidence ›How we check this ›
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