Information on EC 1.12.1.3 - hydrogen dehydrogenase (NADP+)

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The expected taxonomic range for this enzyme is: Bacteria, Archaea, Eukaryota

EC NUMBER
COMMENTARY hide
1.12.1.3
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RECOMMENDED NAME
GeneOntology No.
hydrogen dehydrogenase (NADP+)
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
H2 + NADP+ = H+ + NADPH
show the reaction diagram
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REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
oxidation
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redox reaction
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reduction
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PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
hydrogen oxidation III (anaerobic, NADP)
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hydrogen production IV
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hydrogen production
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SYSTEMATIC NAME
IUBMB Comments
hydrogen:NADP+ oxidoreductase
The protein from the bacterium Desulfovibrio fructosovorans is an iron-sulfur protein that exclusively functions as a hydrogen dehydrogenase [1], while the enzyme from the archaeon Pyrococcus furiosus is a nickel, iron, iron-sulfur protein, that is part of a heterotetrameric complex where the alpha and delta subunits function as a hydrogenase while the beta and gamma subunits function as sulfur reductase (EC 1.12.98.4, sulfhydrogenase). Different from EC 1.12.1.5, hydrogen dehydrogenase [NAD(P)+].
CAS REGISTRY NUMBER
COMMENTARY hide
9027-05-8
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ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
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Manually annotated by BRENDA team
green alga
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Manually annotated by BRENDA team
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
H+ + NADPH
H2 + NADP+
show the reaction diagram
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?
H+ + reduced benzyl viologen
H2 + oxidized benzyl viologen
show the reaction diagram
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?
H2 + 2,6-dichlorophenolindophenol
H+ + ?
show the reaction diagram
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r
H2 + NAD+
H+ + NADH
show the reaction diagram
specific activity with NAD+ as electron acceptor is 150times lower than that of NADP+
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r
H2 + NADP+
H+ + NADPH
show the reaction diagram
H2 + oxidized benzyl viologen
H+ + reduced benzyl viologen
show the reaction diagram
H2 + oxidized ferredoxin
H+ + reduced ferredoxin
show the reaction diagram
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-
?
H2 + oxidized methyl viologen
H+ + reduced methyl viologen
show the reaction diagram
H2 + oxidized methylene blue
H+ + reduced methylene blue
show the reaction diagram
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r
NADPH + H+
H2 + NADP+
show the reaction diagram
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?
additional information
?
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the enzyme introduces corrosion of 316L stainless steel in the presence of NAD+. The enzyme induces the occurrence of cathodic and anodic microsites on the same electrode surface
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NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
H2 + NADP+
H+ + NADPH
show the reaction diagram
H2 + oxidized ferredoxin
H+ + reduced ferredoxin
show the reaction diagram
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?
additional information
?
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the enzyme introduces corrosion of 316L stainless steel in the presence of NAD+. The enzyme induces the occurrence of cathodic and anodic microsites on the same electrode surface
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COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
NAD+
specific activity as electron acceptor is 150times lower than that of NADP+
NADP+
electron acceptor
[Ni-Fe]-center
metal content Ni:Fe is 1:25 for the native enzyme and 1:32 for the recombinant enzyme
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METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Nickel
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
CO
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0.02 mM, 50% inhibition
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.07951
Ferredoxin
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0.012 - 0.02
H2
0.037 - 0.09
NADP+
0.007 - 0.03
NADPH
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.076
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crude extract
87.78
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after affinity chromatogrphy
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
90
H2 production with methyl viologen
95
H2 uptake with methyl viologen
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
PDB
SCOP
CATH
UNIPROT
ORGANISM
Q46505
Desulfovibrio fructosivorans;
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
10920
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truncated HndD subunit, mass spectrometry
11000
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truncated HndA subunit, SDS-PAGE
19000
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HndA subunit, SDS-PAGE
55000
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SDS-PAGE
150000
gel filtration
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
?
(alphabetagammadelta)n, n is 3 or more
heterotetramer
a maturation pathway is proposed where, in the terminal step, the NiFe-containing catalytic subunit assembles with NADPH-oxidizing trimeric form to give the active holoenzym
pentamer
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16.5% tricine-SDS
tetramer
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
80
203 min, 50% loss of activity of evolution reaction
100
t1/2: 2 h, native enzyme
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
50% loss of enzyme activity when extracts are exposed for 15 min to air
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spontaneous proteolysis of the HndA subunit occurs during desalting preparation performed in a Centricon
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OXIDATION STABILITY
ORGANISM
UNIPROT
LITERATURE
t1/2 for the native enzyme is 25 h at 23°C in presence of air, t1/2 for for the recombinant enzyme is 20 h at 23°C in presence of air
746235
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
C-terminal domain of subunit HndA and N-terminal domain of subunit HndD recombinantly expressed in Escherichia coli
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one-step purification of the recombinant enzyme including a polyhistidine affinity tag results in a 50fold increase in yield compared to the four-step purification procedure for the native enzyme
single affinity chromatography, 4°C, aerobic conditions
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strategies for reliable and improved large-scale production of Pyrococcus furiosus with integrated purification of hydrogenase I
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truncated version of HndA subunit
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Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
expression in Escherichia coli. The native Escherichia coli maturation machinery will also generate a functional hydrogenase when provided with only the genes encoding the hydrogenase subunits and a single protease from Pyrococcus furiosus. The expression is induced by anaerobic conditions, whereby Escherichia coli is grown aerobically and production of recombinant hydrogenase is achieved by simply changing the gas feed from air to an inert gas (N2). The recombinant enzyme is purified and shown to be functionally similar to the native enzyme purified from Pyrococcus furiosus. The methodology to generate this key hydrogen-producing enzyme has dramatic implications for the production of hydrogen and NADPH as vehicles for energy storage and transport, for engineering hydrogenase to optimize production and catalysis, as well as for the general production of complex, oxygen-sensitive metalloproteins
expression of C-terminal domain of subunit HndA and N-terminal domain of subunit HndD in Escherichia coli
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HndA and HndB subunit
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HndA and HndD subunit; non-lethal deletions
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overexpression in Synechocystis
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the four gene operon encoding the enzyme is overexpressed in Pyrococcus furiosus and includes a polyhistidine affinity tag
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
energy production
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application in biofuel cells, to generate an electric current
synthesis
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