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Literature summary extracted from

  • Sun, J.; Hopkins, R.; Jenney Jr., F.; McTernan, P.; Adams, M.
    Heterologous expression and maturation of an NADP-Dependent [NiFe]-Hydrogenase A key enzyme in biofuel production (2010), PLoS ONE, 5, e10526 .
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.12.1.3 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 Pyrococcus furiosus

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.12.1.3 0.007
-
 NADPH 80°C, pH 8.4, recombinant enzyme Pyrococcus furiosus
1.12.1.3 0.03
-
 NADPH 80°C, pH 8.4, native enzyme Pyrococcus furiosus

Localization

EC Number Localization Comment Organism GeneOntology No. Textmining
1.12.1.3 cytoplasm
-
 Pyrococcus furiosus 5737
-

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
1.12.1.3 Iron
-
 Pyrococcus furiosus
1.12.1.3 Nickel
-
 Pyrococcus furiosus

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
1.12.1.3 150000
-
 gel filtration Pyrococcus furiosus

Organism

EC Number Organism UniProt Comment Textmining
1.12.1.3 Pyrococcus furiosus E7FI44 AND Q8U2E5 AND Q8U2E4 AND E7FHU4 E7FI44: subunit alpha, Q8U2E5: subunit beta, Q8U2E4: subunit gamma, E7FHU4: subunit delta
-

Oxidation Stability

EC Number Oxidation Stability Organism
1.12.1.3 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 Pyrococcus furiosus

Purification (Commentary)

EC Number Purification (Comment) Organism
1.12.1.3
-
 Pyrococcus furiosus

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.12.1.3 H+ + NADPH
-
 Pyrococcus furiosus H2 + NADP+
-
 ?
1.12.1.3 H+ + reduced benzyl viologen
-
 Pyrococcus furiosus H2 + oxidized benzyl viologen
-
 ?

Synonyms

EC Number Synonyms Comment Organism
1.12.1.3 NADP-dependent hydrogenase
-
 Pyrococcus furiosus
1.12.1.3 SHI
-
 Pyrococcus furiosus
1.12.1.3 soluble hydrogenase I
-
 Pyrococcus furiosus
1.12.1.3 [NiFe]-hydrogenase
-
 Pyrococcus furiosus

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.12.1.3 80
-
 assay at Pyrococcus furiosus

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
1.12.1.3 90
-
 t1/2: 30 h for the native enzyme, t1/2: 1 h for the recombinant enzyme Pyrococcus furiosus
1.12.1.3 100
-
 t1/2: 2 h, native enzyme Pyrococcus furiosus

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.12.1.3 8.4
-
 assay at Pyrococcus furiosus

Cofactor

EC Number Cofactor Comment Organism Structure
1.12.1.3 [Ni-Fe]-center metal content Ni:Fe is 1:25 for the native enzyme and 1:32 for the recombinant enzyme Pyrococcus furiosus