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Spectroscopic and biochemical insight into an electron-bifurcating [FeFe] hydrogenase

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Title: Spectroscopic and biochemical insight into an electron-bifurcating [FeFe] hydrogenase
Authors: Chongdar, Nipa Browse this author
Pawlak, Krzysztof Browse this author
Ruediger, Olaf Browse this author
Reijerse, Edward J. Browse this author
Rodriguez-Macia, Patricia Browse this author
Lubitz, Wolfgang Browse this author
Birrell, James A. Browse this author
Ogata, Hideaki Browse this author →KAKEN DB
Keywords: [FeFe] hydrogenase
Electron bifurcation
Spectroscopy
Electrochemistry
Ferredoxin
Issue Date: Feb-2020
Publisher: Springer
Journal Title: Journal of Biological Inorganic Chemistry
Volume: 25
Issue: 1
Start Page: 135
End Page: 149
Publisher DOI: 10.1007/s00775-019-01747-1
Abstract: The heterotrimeric electron-bifurcating [FeFe] hydrogenase (HydABC) from Thermotoga maritima (Tm) couples the endergonic reduction of protons (H+) by dihydronicotinamide adenine dinucleotide (NADH) ( increment G(0) approximate to 18 kJ mol(-1)) to the exergonic reduction of H+ by reduced ferredoxin (Fd(red)) ( increment G(0) approximate to - 16 kJ mol(-1)). The specific mechanism by which HydABC functions is not understood. In the current study, we describe the biochemical and spectroscopic characterization of TmHydABC recombinantly produced in Escherichia coli and artificially maturated with a synthetic diiron cofactor. We found that TmHydABC catalyzed the hydrogen (H-2)-dependent reduction of nicotinamide adenine dinucleotide (NAD(+)) in the presence of oxidized ferredoxin (Fd(ox)) at a rate of approximate to 17 mu mol NADH min(-1) mg(-1). Our data suggest that only one flavin is present in the enzyme and is not likely to be the site of electron bifurcation. FTIR and EPR spectroscopy, as well as FTIR spectroelectrochemistry, demonstrated that the active site for H-2 conversion, the H-cluster, in TmHydABC behaves essentially the same as in prototypical [FeFe] hydrogenases, and is most likely also not the site of electron bifurcation. The implications of these results are discussed with respect to the current hypotheses on the electron bifurcation mechanism of [FeFe] hydrogenases. Overall, the results provide insight into the electron-bifurcating mechanism and present a well-defined system for further investigations of this fascinating class of [FeFe] hydrogenases. Graphic abstract
Rights: http://creativecommons.org/licenses/by/4.0
Type: article
URI: http://hdl.handle.net/2115/77971
Appears in Collections:低温科学研究所 (Institute of Low Temperature Science) > 雑誌発表論文等 (Peer-reviewed Journal Articles, etc)

Submitter: 緒方 英明

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