ArticlePloS one2010
A cell-free microtiter plate screen for improved [FeFe] hydrogenases.
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 17 papers.
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Who cites it
17 citing papers in PubMed.
- A Gas Chromatography-Based Workflow Leads to the Detection of Hydrogenase Activity in Native Bacteria.Chembiochem : a European journal of chemical biology · 2026Article
- Second and Outer Coordination Sphere Effects in Nitrogenase, Hydrogenase, Formate Dehydrogenase, and CO Dehydrogenase.Chemical reviews · 2022Review
- Synthetic biology for improved hydrogen production in Chlamydomonas reinhardtii.Microbial biotechnology · 2022Review
- Heterologous Hydrogenase Overproduction Systems for Biotechnology-An Overview.International journal of molecular sciences · 2020Review
- Macromolecular crowding effects on transcription and translation are regulated by free magnesium ion.Biotechnology and applied biochemistry · 2020Article
- Article
- Atypical effect of temperature tuning on the insertion of the catalytic iron-sulfur center in a recombinant [FeFe]-hydrogenase.Protein science : a publication of the Protein Society · 2015Article
- Implementation of photobiological H2 production: the O 2 sensitivity of hydrogenases.Photosynthesis research · 2015Article
- [FeFe]-hydrogenase abundance and diversity along a vertical redox gradient in Great Salt Lake, USA.International journal of molecular sciences · 2014Article
- Improvement of biocatalysts for industrial and environmental purposes by saturation mutagenesis.Biomolecules · 2013Article
- Biocatalyst development by directed evolution.Bioresource technology · 2012Review
- Hydrogen production by recombinant Escherichia coli strains.Microbial biotechnology · 2012Review
- Technology prospecting on enzymes: application, marketing and engineering.Computational and structural biotechnology journal · 2012Review
- Article
- A synthetic system links FeFe-hydrogenases to essential E. coli sulfur metabolism.Journal of biological engineering · 2011Article
- Article
- Development of an in vitro compartmentalization screen for high-throughput directed evolution of [FeFe] hydrogenases.PloS one · 2010Article
Corrections and comments
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Authors and funding
2 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
background[FeFe] hydrogenase enzymes catalyze the production and dissociation of H(2), a potential renewable fuel. Attempts to exploit these catalysts in engineered systems have been hindered by the biotechnologically inconvenient properties of the natural enzymes, including their extreme oxygen sensitivity. Directed evolution has been used to improve the characteristics of a range of natural catalysts, but has been largely unsuccessful for [FeFe] hydrogenases because of a lack of convenient screening platforms. METHODOLOGY/PRINCIPAL
findingsHere we describe an in vitro screening technology for oxygen-tolerant and highly active [FeFe] hydrogenases. Despite the complexity of the protocol, we demonstrate a level of reproducibility that allows moderately improved mutants to be isolated. We have used the platform to identify a mutant of the Chlamydomonas reinhardtii [FeFe] hydrogenase HydA1 with a specific activity approximately 4 times that of the wild-type enzyme. CONCLUSIONS/SIGNIFICANCE: Our results demonstrate the feasibility of using the screen presented here for large-scale efforts to identify improved biocatalysts for energy applications. The system is based on our ability to activate these complex enzymes in E. coli cell extracts, which allows unhindered access to the protein maturation and assay environment.
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