ArticlePloS one2010
High-yield expression of heterologous [FeFe] hydrogenases in Escherichia coli.
Article in PloS one, 2010. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 71 papers.
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Who cites it
71 citing papers in PubMed, 178 citations in OpenAlex.
- The anaerobic fungusmBio · 2026Article
- Mössbauer spectroscopy in drug discovery: revealing Fe- and Fe-S cluster dependent targets.Expert opinion on drug discovery · 2026Review
- A Low-Potential π‑Extended Viologen Electron Donor Results in Increased HACS organic & inorganic Au · 2026Article
- Heterologous production of tungsten-dependent formate dehydrogenase I from Methylorubrum extorquens in Escherichia coli reveals α-subunit maturation as the major bottleneck.Bioresources and bioprocessing · 2026Article
- Photoactivation of [FeFe] Hydrogenase Studied by Multiscale Time-Resolved Infrared Spectroscopy.The journal of physical chemistry letters · 2026Article
- Clostridium beijerinckii displays a soluble [FeFe]-hydrogenase/formate dehydrogenase enzyme complex that links H2 and CO2 metabolism.The Biochemical journal · 2026Article
- Reconsidering the Enzyme Kinetics of [FeFe]-Hydrogenases: Improved Turnover Rates and New Insights into pH and Potential Dependence with Eu(II)-Based Solution Assays.Analytical chemistry · 2025Article
- In vitro maturation of fully active [FeFe]-hydrogenase in a defined system including the iron carrier NfuA.Proceedings of the National Academy of Sciences of the United States of America · 2025Article
- Hydrogen-Producing Catalysts Based on Ferredoxin Scaffolds.Advanced science (Weinheim, Baden-Wurttemberg, Germany) · 2025Article
- Structural determinants of oxygen resistance and ZnProceedings of the National Academy of Sciences of the United States of America · 2025Article
- Using Phylogeny and a Conserved Genomic Neighborhood Analysis to Extract and Visualize Gene Sets Involved in Target Gene Function: The Case of [NiFe]-hydrogenase and Succinate Dehydrogenase.Microbes and environments · 2025Article
- Computation-guided transcription factor biosensor specificity engineering for adipic acid detection.Computational and structural biotechnology journal · 2024Article
- Bioelectrocatalytic carbon dioxide reduction by an engineered formate dehydrogenase from Thermoanaerobacter kivui.Nature communications · 2024Article
- Ammonia synthesis via an engineered nitrogenase assembly pathway inNature catalysis · 2024Article
- Conjugated Polymer/RecombinantACS nano · 2024Article
- Kinetic Modeling of the Reversible or Irreversible Electrochemical Responses of FeFe-Hydrogenases.Journal of the American Chemical Society · 2024Article
- The AlgaInternational journal of molecular sciences · 2023Article
- Molecular architecture and electron transfer pathway of the Stn family transhydrogenase.Nature communications · 2023Article
- Maturation strategy influences expression levels and cofactor occupancy in Fe-S proteins.Journal of biological inorganic chemistry : JBIC : a publication of the Society of Biological Inorganic Chemistry · 2023Article
- Increasing the OACS catalysis · 2023Article
11 more citing papers are in PubMed but not listed here.
Corrections and comments
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Authors and funding
6 authors at 3 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
backgroundThe realization of hydrogenase-based technologies for renewable H(2) production is presently limited by the need for scalable and high-yielding methods to supply active hydrogenases and their required maturases. PRINCIPAL
findingsIn this report, we describe an improved Escherichia coli-based expression system capable of producing 8-30 mg of purified, active [FeFe] hydrogenase per liter of culture, volumetric yields at least 10-fold greater than previously reported. Specifically, we overcame two problems associated with other in vivo production methods: low protein yields and ineffective hydrogenase maturation. The addition of glucose to the growth medium enhances anaerobic metabolism and growth during hydrogenase expression, which substantially increases total yields. Also, we combine iron and cysteine supplementation with the use of an E. coli strain upregulated for iron-sulfur cluster protein accumulation. These measures dramatically improve in vivo hydrogenase activation. Two hydrogenases, HydA1 from Chlamydomonas reinhardtii and HydA (CpI) from Clostridium pasteurianum, were produced with this improved system and subsequently purified. Biophysical characterization and FTIR spectroscopic analysis of these enzymes indicate that they harbor the H-cluster and catalyze H(2) evolution with rates comparable to those of enzymes isolated from their respective native organisms. SIGNIFICANCE: The production system we describe will facilitate basic hydrogenase investigations as well as the development of new technologies that utilize these prolific H(2)-producing enzymes. These methods can also be extended for producing and studying a variety of oxygen-sensitive iron-sulfur proteins as well as other proteins requiring anoxic environments.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.