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Literature summary extracted from

  • Schneider, K.; Schlegel, H.G.
    Purification and properties of soluble hydrogenase from Alcaligenes eutrophus H 16 (1976), Biochim. Biophys. Acta, 452, 66-80.
    View publication on PubMed

General Stability

EC Number General Stability Organism
1.12.1.2 unstable in reduced active form Cupriavidus necator

Inhibitors

EC Number Inhibitors Comment Organism Structure
1.12.1.2 high ionic strength 200 mM phosphate buffer, 70% inhibition Cupriavidus necator

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.12.1.2 0.037
-
 H2
-
 Cupriavidus necator
1.12.1.2 0.123
-
 NAD+
-
 Cupriavidus necator
1.12.1.2 0.56
-
 NAD+
-
 Cupriavidus necator

Localization

EC Number Localization Comment Organism GeneOntology No. Textmining
1.12.1.2 soluble
-
 Cupriavidus necator
-
-

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
1.12.1.2 Iron non-heme iron protein Cupriavidus necator
1.12.1.2 Iron no increase of activity by addition of Co2+, Mn2+, Ni2+ or Fe2+ Cupriavidus necator
1.12.1.2 additional information no increase of activity by addition of Co2+, Mn2+, Ni2+ or Fe2+ Cupriavidus necator

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
1.12.1.2 204500
-
 gel filtration Cupriavidus necator
1.12.1.2 206000
-
 sucrose density gradient centrifugation Cupriavidus necator

Organism

EC Number Organism UniProt Comment Textmining
1.12.1.2 Cupriavidus necator
-
-
-
1.12.1.2 Cupriavidus necator
-
 formerly Alcaligenes eutrophus
-
1.12.1.2 Cupriavidus necator H16 / ATCC 23440 / NCIB 10442 / S-10-1
-
-
-

Oxidation Stability

EC Number Oxidation Stability Organism
1.12.1.2 oxidized hydrogenase purified under aerobic conditions is highly stable but not reactive, reductive activation of the enzyme by H2 in the presence of catalytic amounts of NADH or by reducing agents destabilizes Cupriavidus necator

Purification (Commentary)

EC Number Purification (Comment) Organism
1.12.1.2 sp. H16, cetavlon, ammonium sulfate, DEAE-cellulose, Sephadex G-200, hydroxyapatite Cupriavidus necator

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
1.12.1.2 54.46
-
-
 Cupriavidus necator

Storage Stability

EC Number Storage Stability Organism
1.12.1.2 -20°C, O2, several months, no loss in activity Cupriavidus necator

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.12.1.2 2 ferricyanide + NAD(P)H
-
 Cupriavidus necator 2 ferrocyanide + NAD(P)+ + H+
-
 ?
1.12.1.2 H2 + NAD+ electron acceptor FMN Cupriavidus necator H+ + NADH
-
 r
1.12.1.2 H2 + NAD+ electron acceptor FAD Cupriavidus necator H+ + NADH
-
 r
1.12.1.2 H2 + NAD+ artificial electron acceptors: Janus green, 2,6-dichlorophenolindophenol, phenazine methosulfate, menaquinone, ubiquinone, cytochrome c Cupriavidus necator H+ + NADH
-
 r
1.12.1.2 H2 + NAD+ artificial electron acceptor benzyl viologen Cupriavidus necator H+ + NADH
-
 r
1.12.1.2 H2 + NAD+ artificial electron acceptor methyl viologen Cupriavidus necator H+ + NADH
-
 r
1.12.1.2 H2 + NAD+ weak electron acceptor O2 Cupriavidus necator H+ + NADH
-
 r
1.12.1.2 H2 + NAD+ enzyme also has diaphorase and NAD(P)H oxidase activity Cupriavidus necator H+ + NADH
-
 r
1.12.1.2 H2 + NAD+ electron acceptor NAD+ Cupriavidus necator H+ + NADH
-
 r
1.12.1.2 H2 + NAD+ artificial electron acceptor ferricyanide Cupriavidus necator H+ + NADH
-
 r
1.12.1.2 H2 + NAD+ artificial electron acceptor methylene blue Cupriavidus necator H+ + NADH
-
 r
1.12.1.2 H2 + NAD+ electron acceptor FMN Cupriavidus necator H16 / ATCC 23440 / NCIB 10442 / S-10-1 H+ + NADH
-
 r
1.12.1.2 H2 + NAD+ electron acceptor FAD Cupriavidus necator H16 / ATCC 23440 / NCIB 10442 / S-10-1 H+ + NADH
-
 r
1.12.1.2 H2 + NAD+ artificial electron acceptors: Janus green, 2,6-dichlorophenolindophenol, phenazine methosulfate, menaquinone, ubiquinone, cytochrome c Cupriavidus necator H16 / ATCC 23440 / NCIB 10442 / S-10-1 H+ + NADH
-
 r
1.12.1.2 H2 + oxidized benzyl viologen
-
 Cupriavidus necator H+ + reduced benzyl viologen
-
 r
1.12.1.2 H2 + oxidized methyl viologen
-
 Cupriavidus necator H+ + reduced methyl viologen
-
 r
1.12.1.2 H2 + oxidized methyl viologen
-
 Cupriavidus necator H16 / ATCC 23440 / NCIB 10442 / S-10-1 H+ + reduced methyl viologen
-
 r
1.12.1.2 NAD(P)H + H+ + oxidized 2,6-dichlorophenolindophenol
-
 Cupriavidus necator NAD(P)+ + reduced 2,6-dichlorophenolindophenol
-
 ?
1.12.1.2 NAD(P)H + H+ + oxidized 2,6-dichlorophenolindophenol
-
 Cupriavidus necator H16 / ATCC 23440 / NCIB 10442 / S-10-1 NAD(P)+ + reduced 2,6-dichlorophenolindophenol
-
 ?
1.12.1.2 oxidized cytochrome c + NAD(P)H
-
 Cupriavidus necator reduced cytchrome c + NAD(P)+
-
 ?
1.12.1.2 oxidized methylene blue + NAD(P)H
-
 Cupriavidus necator reduced methylene blue + NAD(P)+
-
 ?

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.12.1.2 33
-
-
 Cupriavidus necator

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.12.1.2 8
-
-
 Cupriavidus necator

Cofactor

EC Number Cofactor Comment Organism Structure
1.12.1.2 ATP 30% stimulation Cupriavidus necator
1.12.1.2 flavin flavoprotein Cupriavidus necator
1.12.1.2 FMN 25% stimulation Cupriavidus necator