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Spectroscopic investigations under whole cell conditions provide new insight into the metal hydride chemistry of [FeFe]-hydrogenase

revised on 28.01.2020, 13:37 and posted on 29.01.2020, 05:29 by Lívia S. Mészáros, Pierre Ceccaldi, Marco Lorenzi, Holly J. Redman, Emanuel Pfitzner, Joachim Heberle, Moritz Senger, Sven T. Stripp, Gustav Berggren
Hydrogenases are among the fastest H2 evolving catalysts known to date and have been extensively studied under in vitro conditions. Here, we report the first mechanistic investigation of an [FeFe]-hydrogenase under in vivo conditions. Functional [FeFe]-hydrogenase from the green alga Chlamydomonas reinhardtii is generated in genetically modified Escherichia coli cells, by addition of a synthetic cofactor to the growth medium. The assembly and reactivity of the resulting semi-synthetic enzyme was monitored using whole-cell electron paramagnetic resonance as well as Fourier-transform infrared spectroscopy. Through a combination of gas treatments, pH titrations and isotope editing, we were able to corroborate the physiological relevance of a number of proposed catalytic intermediates, including reactive iron-hydride species. We demonstrate the formation of the so-called hydride state in vivo. Moreover, two previously uncharacterized redox species are reported herein, illustrating the complex metal hydride chemistry of [FeFe]-hydrogenase.


Olle Engkvist Byggmästare Foundation

European Research Council

Deutsche Forschungsgemeinschaft

EXC 2008:  UniSysCat

Deutsche Forschungsgemeinschaft

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Freie Universität Berlin



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No conflict of interest.