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Information on EC 1.12.1.4 - hydrogenase (NAD+, ferredoxin) for references in articles please use BRENDA:EC1.12.1.4Word Map on EC 1.12.1.4
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The expected taxonomic range for this enzyme is: Bacteria
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hydrogenase (NAD+, ferredoxin)
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2 H2 + NAD+ + 2 oxidized ferredoxin = 5 H+ + NADH + 2 reduced ferredoxin
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hydrogen production I
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hydrogen:NAD+, ferredoxin oxidoreductase
The enzyme from Thermotoga maritima contains a [FeFe] cluster (H-cluster) and iron-sulfur clusters. It works in the direction evolving hydrogen as a means of eliminating excess reducing equivalents.
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bifurcating [FeFe] hydrogenase
electron-bifurcating ferredoxin- and NAD-dependent hydrogenase
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NAD(P)+-linked [FeFe]-hydrogenase
[FeFe]-type hydrogenase
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bifurcating [FeFe] hydrogenase
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bifurcating [FeFe] hydrogenase
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Fe-only hydrogenase
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Fe-only hydrogenase
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HydABC
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HydC
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gamma subunit
NAD(P)+-linked [FeFe]-hydrogenase
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NAD(P)+-linked [FeFe]-hydrogenase
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physiological function
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an active HydF GTPase domain is essential to produce a functional [FeFe]-hydrogenase
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H2 + NAD+ + oxidized ferredoxin
H+ + NADH + reduced ferredoxin
NAD+ + 2 oxidized ferredoxin + 2 H2
NADH + 3 H+ + 2 reduced ferredoxin
NAD+ + 2 oxidized methyl viologen + 2 H2
NADH + 3 H+ + 2 reduced methyl viologen
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NADH + 3 H+ + 2 reduced ferredoxin
NAD+ + 2 oxidized ferredoxin + 2 H2
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per mol NADH, about 2 mol H2 is formed
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NADH + 3 H+ + 2 reduced methyl viologen
NAD+ + 2 oxidized methyl viologen + 2 H2
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NADH + H+ + reduced ferredoxin
H2 + NAD+ + oxidized ferredoxin
NADH + H+ + reduced methyl viologen
H2 + NAD+ + oxidized methyl viologen
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sodium dithionite + H+ + reduced anthraquinone-2,6-disulfonic acid
H2 + oxidized anthraquinone-2,6-disulfonic acid + ?
sodium dithionite + H+ + reduced methyl viologen
H2 + oxidized methyl viologen + ?
H2 + NAD+ + oxidized ferredoxin
H+ + NADH + reduced ferredoxin
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H2 + NAD+ + oxidized ferredoxin
H+ + NADH + reduced ferredoxin
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NAD+ + 2 oxidized ferredoxin + 2 H2
NADH + 3 H+ + 2 reduced ferredoxin
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ferredoxin reduction with H2 is driven by the exergonic reduction of NAD+ (E0 -320 mV) with H2. In the absence of NAD+, ferredoxin is not reduced
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NAD+ + 2 oxidized ferredoxin + 2 H2
NADH + 3 H+ + 2 reduced ferredoxin
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NAD+ + 2 oxidized ferredoxin + 2 H2
NADH + 3 H+ + 2 reduced ferredoxin
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NADH + H+ + reduced ferredoxin
H2 + NAD+ + oxidized ferredoxin
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NADH + H+ + reduced ferredoxin
H2 + NAD+ + oxidized ferredoxin
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the iron hydrogenase requires the presence of both electron carriers, NADH and ferredoxin, for catalysis of H2 production
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NADH + H+ + reduced ferredoxin
H2 + NAD+ + oxidized ferredoxin
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the iron hydrogenase requires the presence of both electron carriers, NADH and ferredoxin, for catalysis of H2 production
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NADH + H+ + reduced ferredoxin
H2 + NAD+ + oxidized ferredoxin
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sodium dithionite + H+ + reduced anthraquinone-2,6-disulfonic acid
H2 + oxidized anthraquinone-2,6-disulfonic acid + ?
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sodium dithionite + H+ + reduced anthraquinone-2,6-disulfonic acid
H2 + oxidized anthraquinone-2,6-disulfonic acid + ?
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sodium dithionite + H+ + reduced methyl viologen
H2 + oxidized methyl viologen + ?
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sodium dithionite + H+ + reduced methyl viologen
H2 + oxidized methyl viologen + ?
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H2 + NAD+ + oxidized ferredoxin
H+ + NADH + reduced ferredoxin
NADH + H+ + reduced ferredoxin
H2 + NAD+ + oxidized ferredoxin
H2 + NAD+ + oxidized ferredoxin
H+ + NADH + reduced ferredoxin
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H2 + NAD+ + oxidized ferredoxin
H+ + NADH + reduced ferredoxin
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NADH + H+ + reduced ferredoxin
H2 + NAD+ + oxidized ferredoxin
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NADH + H+ + reduced ferredoxin
H2 + NAD+ + oxidized ferredoxin
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the iron hydrogenase requires the presence of both electron carriers, NADH and ferredoxin, for catalysis of H2 production
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NADH + H+ + reduced ferredoxin
H2 + NAD+ + oxidized ferredoxin
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the iron hydrogenase requires the presence of both electron carriers, NADH and ferredoxin, for catalysis of H2 production
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NADH + H+ + reduced ferredoxin
H2 + NAD+ + oxidized ferredoxin
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4Fe-4S-center
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displays electron paramagnetic resonance signals characteristic of oxidized [3Fe-4S]+ cluster. Cysteine residues C302, C353, and C356 are strictly required for the binding of the [4Fe-4S] cluster, whereas the fourth ligand of the coordination sphere can vary depending on the molecular environment created by local residues and/or experimental conditions
additional information
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no cofactor: NADP+
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FMN
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FMN
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purified enzyme contains 31 irons and 0.8 FMN per heterotrimer
iron-sulfur centre
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the alpha subunit contains two [2Fe-2S] clusters and three [4Fe-4S] clusters, the beta subunit contains three [4Fe-4S] clusters and one [2Fe-2S] clusters, the gamma subunit contains one [2Fe-2S] cluster
iron-sulfur centre
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subunit HydA (63 kDa) is predicted to harbor the H cluster, i.e., the [6Fe4S] hydrogenase active site, three [4Fe4S] clusters, and one [2Fe2S] cluster. Subunit HydB (67 kDa) is predicted to harbor three [4Fe4S] clusters, a flavin binding site, and an NAD binding site. HydC (17 kDa) is predicted to harbor one [2Fe2S] cluster
NAD+
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NADH
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Fe2+
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the holoprotein contains 31.1 mol iron per mol enzyme
Iron
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purified enzyme contains 31 irons and 0.8 FMN per heterotrimer
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additional information
H2
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apparent Km value for H2 is about 6% in the gas phase, pH 7.5, 45°C
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4.6
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oxidation of ferredoxin, pH 7.5, 45°C
57.5
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reduction of ferredoxin, pH 7.5, 45°C
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reduction of methyl viologen, pH 7.5, 45°C
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18025
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1 * 72248 + 1 * 68676 + 1 * 18025, calculated from amino acid sequence
19000
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1 * 73000 + 1 * 68000 + 1 * 19000, SDS-PAGE
68676
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1 * 72248 + 1 * 68676 + 1 * 18025, calculated from amino acid sequence
72248
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1 * 72248 + 1 * 68676 + 1 * 18025, calculated from amino acid sequence
73000
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1 * 73000 + 1 * 68000 + 1 * 19000, SDS-PAGE
160000
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calculated from amino acid sequence
68000
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1 * 73000 + 1 * 68000 + 1 * 19000, SDS-PAGE
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heterohexamer
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2 * 67000, subunit HydB, plus 2 * 63000, subunit HydA, plus 2 * 17000, subunit HydC, SDS-PAGE
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x * 25000, subunit HydF, SDS-PAGE
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x * 50000, subunit HydF, SDS-PAGE
heterotrimer
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heterotrimer
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1 * 72248 + 1 * 68676 + 1 * 18025, calculated from amino acid sequence; 1 * 73000 + 1 * 68000 + 1 * 19000, SDS-PAGE
heterotrimer
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; 1 * 72248 + 1 * 68676 + 1 * 18025, calculated from amino acid sequence; 1 * 73000 + 1 * 68000 + 1 * 19000, SDS-PAGE
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multimer
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x * 68000, subunit HdB, plus x * 64000, subunit HydA, plus x * 18000, subunit HydC
multimer
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x * 68000, subunit HdB, plus x * 64000, subunit HydA, plus x * 18000, subunit HydC
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DEAE-Sepharose column chromatography, Q-Sepharose column chromatography, hydroxyapatite column chromatography, phenyl-Sepharose column chromatography, and Superdex 200 gel filtration
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nickel affinity column chromatography and Superdex 200 gel filtration
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nickel affinity column chromatography, and Superose 12 gel filtration
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streptactin column chromatography
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under strictly anoxic conditions, the enzyme is inactivated in the presence of even trace amounts of O2
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expressed in Escherichia coli BL21(DE3) cells
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expressed in Escherichia coli Rosetta (DE3) cells
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expression in Escherichia coli
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A85P
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the mutant has an altered reduction potential compared to the wild type enzyme
V131N
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the mutant has an altered reduction potential compared to the wild type enzyme
A85P
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the mutant has an altered reduction potential compared to the wild type enzyme
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V131N
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the mutant has an altered reduction potential compared to the wild type enzyme
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C302S
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mutant displays a very weak electron paramagnetic resonance signal
C353S
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mutant displays a very weak electron paramagnetic resonance signal
C356S
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mutant displays a very weak electron paramagnetic resonance signal
H304A
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similar to wild-type, mutant exhibits electron paramagnetic resonance signals characteristic of oxidized [3Fe-4S]+ cluster
H352A
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similar to wild-type, mutant exhibits electron paramagnetic resonance signals characteristic of oxidized [3Fe-4S]+ cluster
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A0A0P0LLL7_BACVU
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8939
TrEMBL
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Schut, G.J.; Adams, M.W.
The iron-hydrogenase of Thermotoga maritima utilizes ferredoxin and NADH synergistically: a new perspective on anaerobic hydrogen production
J. Bacteriol.
191
4451-4457
2009
Thermotoga maritima, Thermotoga maritima DSM 3109
brenda
Verhagen, M.; Adams, M.
Fe-only hydrogenase from Thermotoga maritima
Methods Enzymol.
331
216-226
2001
Thermotoga maritima, Thermotoga maritima DSM 3109
brenda
Berto, P.; Di Valentin, M.; Cendron, L.; Vallese, F.; Albertini, M.; Salvadori, E.; Giacometti, G.M.; Carbonera, D.; Costantini, P.
The [4Fe-4S]-cluster coordination of [FeFe]-hydrogenase maturation protein HydF as revealed by EPR and HYSCORE spectroscopies
Biochim. Biophys. Acta
1817
2149-2157
2012
Thermotoga neapolitana
brenda
Wang, S.; Huang, H.; Kahnt, J.; Thauer, R.K.
A reversible electron-bifurcating ferredoxin- and NAD-dependent [FeFe]-hydrogenase (HydABC) in Moorella thermoacetica
J. Bacteriol.
195
1267-1275
2013
Moorella thermoacetica
brenda
Zheng, Y.; Kahnt, J.; Kwon, I.H.; Mackie, R.I.; Thauer, R.K.
Hydrogen formation and its regulation in Ruminococcus albus: Involvement of an electron-bifurcating [FeFe]-hydrogenase, of a non-electron-bifurcating [FeFe]-hydrogenase, and of a putative hydrogen-sensing [FeFe]-hydrogenase
J. Bacteriol.
196
3840-3852
2014
Ruminococcus albus, Ruminococcus albus DSM 20455
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Maso, L.; Galazzo, L.; Vallese, F.; Di Valentin, M.; Albertini, M.; De Rosa, E.; Giacometti, G.; Costantini, P.; Carbonera, D.
A conformational study of the GTPase domain of [FeFe]-hydrogenase maturation protein HydF by PELDOR spectroscopy
Appl. Magn. Reson.
46
465-479
2015
Thermotoga neapolitana
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brenda
Birrell, J.A.; Laurich, C.; Reijerse, E.J.; Ogata, H.; Lubitz, W.
Importance of hydrogen bonding in fine tuning the [2Fe-2S] cluster redox potential of HydC from Thermotoga maritima
Biochemistry
55
4344-4355
2016
Thermotoga maritima, Thermotoga maritima MSB8 / DSM 3109 / ATCC 43589
brenda
Albertini, M.; Vallese, F.; Di Valentin, M.; Berto, P.; Giacometti, G.; Costantini, P.; Carbonera, D.
The proton iron-sulfur cluster environment of the [FeFe]-hydrogenase maturation protein HydF from Thermotoga neapolitana
Int. J. Hydrogen Energy
39
18574-18582
2014
Thermotoga neapolitana
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brenda
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