Expression of the [FeFe] hydrogenase in the chloroplast of Chlamydomonas reinhardtii

Biological hydrogen generation from phototrophic organisms is a promising source of renewable fuel. The nuclear-expressed [FeFe] hydrogenase from Chlamydomonas reinhardtii has an extremely high turnover rate, and so has been a target of intense research. Here, we demonstrate that a codon-optimized n...

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Bibliographic Details
Published in:International journal of hydrogen energy 2014-03, Vol.39 (8), p.3657-3665
Main Authors: Reifschneider-Wegner, Kiera, Kanygin, Andrey, Redding, Kevin E.
Format: Article
Language:English
Subjects:
[FeFe] hydrogenase
Alternative fuels. Production and utilization
Applied sciences
Chlamydomonas
Chlamydomonas reinhardtii
Chloroplast
Energy
Exact sciences and technology
Fuels
Gene expression
Green algae
Hydrogen
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Summary:Biological hydrogen generation from phototrophic organisms is a promising source of renewable fuel. The nuclear-expressed [FeFe] hydrogenase from Chlamydomonas reinhardtii has an extremely high turnover rate, and so has been a target of intense research. Here, we demonstrate that a codon-optimized native hydrogenase can be successfully expressed in the chloroplast. We also demonstrate a curiously strong negative selective pressure resulting from unregulated hydrogenase expression in this location, and discuss management of its expression with a vitamin-controlled gene repression system. To the best of our knowledge, this represents the first example of a nuclear-expressed, chloroplast-localized metalloprotein being synthesized in situ. Control of this process opens up several bioengineering possibilities for the production of biohydrogen. •We express an active [FeFe] hydrogenase in the chloroplast of Chlamydomonas reinhardtii.•The chloroplast hydrogenase is no longer regulated by the native control system.•Production of the apoprotein occurs under both aerobic and anaerobic conditions.•Unregulated expression results in a strong negative selective pressure.•A vitamin-controlled gene repression system suitably manages the negative effects.
ISSN:0360-3199
1879-3487
DOI:10.1016/j.ijhydene.2013.12.157