ArticleThe Journal of biological chemistry2012
Engineering hyperthermophilic archaeon Pyrococcus furiosus to overproduce its cytoplasmic [NiFe]-hydrogenase.
Article in The Journal of biological chemistry, 2012. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 21 papers.
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21 citing papers in PubMed, 53 citations in OpenAlex.
- An unprecedented function for a tungsten-containing oxidoreductase.Journal of biological inorganic chemistry : JBIC : a publication of the Society of Biological Inorganic Chemistry · 2022Article
- Encapsulins.Annual review of biochemistry · 2022Review
- Large-scale computational discovery and analysis of virus-derived microbial nanocompartments.Nature communications · 2021Article
- Methanosarcina acetivorans contains a functional ISC system for iron-sulfur cluster biogenesis.BMC microbiology · 2020Article
- Improving Arsenic Tolerance of Pyrococcus furiosus by Heterologous Expression of a Respiratory Arsenate Reductase.Applied and environmental microbiology · 2020Article
- Characterization of thiosulfate reductase from Pyrobaculum aerophilum heterologously produced in Pyrococcus furiosus.Extremophiles : life under extreme conditions · 2020Article
- Distinct Physiological Roles of the Three Ferredoxins Encoded in the Hyperthermophilic ArchaeonmBio · 2019Article
- Characterization of membrane-bound sulfane reductase: A missing link in the evolution of modern day respiratory complexes.The Journal of biological chemistry · 2018Article
- Biotechnology of extremely thermophilic archaea.FEMS microbiology reviews · 2018Review
- Enzymatic and spectroscopic properties of a thermostable [NiFe]‑hydrogenase performing HBiochimica et biophysica acta. Bioenergetics · 2018Article
- Docking and migration of carbon monoxide in nitrogenase: the case for gated pockets from infrared spectroscopy and molecular dynamics.Biochemistry · 2015Article
- High-yield hydrogen production from biomass by in vitro metabolic engineering: Mixed sugars coutilization and kinetic modeling.Proceedings of the National Academy of Sciences of the United States of America · 2015Article
- Production and Application of a Soluble Hydrogenase from Pyrococcus furiosus.Archaea (Vancouver, B.C.) · 2015Review
- Engineering the respiratory membrane-bound hydrogenase of the hyperthermophilic archaeon Pyrococcus furiosus and characterization of the catalytically active cytoplasmic subcomplex.Protein engineering, design & selection : PEDS · 2015Article
- Intact functional fourteen-subunit respiratory membrane-bound [NiFe]-hydrogenase complex of the hyperthermophilic archaeon Pyrococcus furiosus.The Journal of biological chemistry · 2014Article
- Engineering hydrogen gas production from formate in a hyperthermophile by heterologous production of an 18-subunit membrane-bound complex.The Journal of biological chemistry · 2014Article
- Exploiting microbial hyperthermophilicity to produce an industrial chemical, using hydrogen and carbon dioxide.Proceedings of the National Academy of Sciences of the United States of America · 2013Article
- Genome sequencing of a genetically tractable Pyrococcus furiosus strain reveals a highly dynamic genome.Journal of bacteriology · 2012Article
- Recombinogenic properties of Pyrococcus furiosus strain COM1 enable rapid selection of targeted mutants.Applied and environmental microbiology · 2012Article
- Overview of the genetic tools in the Archaea.Frontiers in microbiology · 2012Article
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Authors and funding
6 authors at 2 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
The cytoplasmic hydrogenase (SHI) of the hyperthermophilic archaeon Pyrococcus furiosus is an NADP(H)-dependent heterotetrameric enzyme that contains a nickel-iron catalytic site, flavin, and six iron-sulfur clusters. It has potential utility in a range of bioenergy systems in vitro, but a major obstacle in its use is generating sufficient amounts. We have engineered P. furiosus to overproduce SHI utilizing a recently developed genetic system. In the overexpression (OE-SHI) strain, transcription of the four-gene SHI operon was under the control of a strong constitutive promoter, and a Strep-tag II was added to the N terminus of one subunit. OE-SHI and wild-type P. furiosus strains had similar rates of growth and H(2) production on maltose. Strain OE-SHI had a 20-fold higher transcription of the polycistronic hydrogenase mRNA encoding SHI, and the specific activity of the cytoplasmic hydrogenase was ∼10-fold higher when compared with the wild-type strain, although the expression levels of genes encoding processing and maturation of SHI were the same in both strains. Overexpressed SHI was purified by a single affinity chromatography step using the Strep-tag II, and it and the native form had comparable activities and physical properties. Based on protein yield per gram of cells (wet weight), the OE-SHI strain yields a 100-fold higher amount of hydrogenase when compared with the highest homologous [NiFe]-hydrogenase system previously reported (from Synechocystis). This new P. furiosus system will allow further engineering of SHI and provide hydrogenase for efficient in vitro biohydrogen production.
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