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Literature summary for 1.12.1.2 extracted from

  • Van der Linden, E.; Burgdorf, T.; Bernhard, M.; Bleijlevens, B.; Friedrich, B.; Albracht, S.P.
    The soluble [NiFe]-hydrogenase from Ralstonia eutropha contains four cyanides in its active site, one of which is responsible for the insensitivity towards oxygen (2004), J. Biol. Inorg. Chem., 9, 616-626.
    View publication on PubMed
Search for more literature for 1.12.1.2

Activating Compound

Activating Compound Comment Organism Structure
FMN
-
 Cupriavidus necator

Inhibitors

Inhibitors Comment Organism Structure
additional information release of one FMN reduces the NAD+ reduction by 90%, but is reversible by addition of excess FMN, insensitivity towards oxygen, with all 4 CN- groups bound to the enzyme, and towards CO Cupriavidus necator
O2 irreversible enzyme inhibition by oxygen occurs if the CN- bound to Ni2+ is irreversibly removed or if the enzyme is reduced by NADH Cupriavidus necator

Localization

Localization Comment Organism GeneOntology No. Textmining
cytoplasm
-
 Cupriavidus necator 5737
-
soluble
-
 Cupriavidus necator
-
-

Metals/Ions

Metals/Ions Comment Organism Structure
CN- enzyme contains four cyanides in its active site, the Ni2+ bound one is responsible for the insensitivity towards oxygen, the active site is a (enzyme-Cys)2(CN)Ni(micro-enzyme-Cys)2Fe(CN)3(CO) centre, the CN- bound to the nickel ion can be irreversibly removed inducing enzyme inhibition by oxygen Cupriavidus necator
CO bound to the active site, the active site is a (enzyme-Cys)2(CN)Ni(micro-enzyme-Cys)2Fe(CN)3(CO) centre Cupriavidus necator
cyanide enzyme contains four cyanides in its active site, one of which is responsible for the insensitivity towards oxygen Cupriavidus necator
Fe2+ enzyme contains a [Ni-Fe] cluster Cupriavidus necator
Fe2+ the active site is a (enzyme-Cys)2(CN)Ni(micro-enzyme-Cys)2Fe(CN)3(CO) centre Cupriavidus necator
Ni2+ enzyme contains a [Ni-Fe] cluster Cupriavidus necator
Ni2+ the active site is a (enzyme-Cys)2(CN)Ni(micro-enzyme-Cys)2Fe(CN)3(CO) centre, H2 activation solely takes place on Ni2+ Cupriavidus necator

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
H2 + NAD+ Cupriavidus necator
-
 H+ + NADH
-
 ?
H2 + NAD+ Cupriavidus necator
-
 H+ + NADH
-
 r

Organism

Organism UniProt Comment Textmining
Cupriavidus necator
-
-
-
Cupriavidus necator
-
 facultative lithoautotrophic Knallgas bacterium
-

Oxidation Stability

Oxidation Stability Organism
insensitive to O2 Cupriavidus necator
insensitivity towards oxygen, irreversible enzyme inhibition by oxygen occurs if the CN- bound to Ni2+ is irreversibly removed or if the enzyme is reduced by NADH Cupriavidus necator

Purification (Commentary)

Purification (Comment) Organism
-
 Cupriavidus necator

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
additional information
-
-
 Cupriavidus necator
17 84 activity of different batches with NAD+ and H2 Cupriavidus necator
30 100 purified enzyme, forward reaction under aerobic conditions with NAD+ Cupriavidus necator
125 175 activity of different batches with NADH and ferricyanide Cupriavidus necator
1263
-
 activity of different batches with benzyl viologen and H2 Cupriavidus necator

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
ferrocyanide + NAD+
-
 Cupriavidus necator ferricyanide + NADH
-
 r
ferrocyanide + NAD+ diaphorase activity Cupriavidus necator ferricyanide + NADH
-
 r
H2 + NAD+
-
 Cupriavidus necator H+ + NADH
-
 ?
H2 + NAD+
-
 Cupriavidus necator H+ + NADH
-
 r
H2 + NAD+ H2 activation solely takes place on Ni2+ Cupriavidus necator H+ + NADH
-
 r
H2 + oxidized benzyl viologen
-
 Cupriavidus necator H+ + reduced benzyl viologen
-
 r
H2 + oxidized benzyl viologen hydrogenase activity Cupriavidus necator H+ + reduced benzyl viologen
-
 r
additional information addition of NADH prolonged the lag phase before H2 consumption Cupriavidus necator ?
-
 ?
additional information FMN release induces reduction with NADH, enzyme shows both hydrogenase and diaphorase activities, proton channeling Cupriavidus necator ?
-
 ?

Subunits

Subunits Comment Organism
More subunits HoxHY form a heterodimer which is responsible for the hydrogenase activity, subunits HoxFU form a heterodimer which is responsible for the NADH-dehydrogenase or diaphorase activity Cupriavidus necator
tetramer enzyme is composed of 4 Hox subunits, HoxF, HoxH, HoxU, and HoxY, with MWs of 67 kDa, 55 kDa, 26 kDa, and 23 kDa Cupriavidus necator

Synonyms

Synonyms Comment Organism
NAD+-reducing [NiFe]-hydrogenase
-
 Cupriavidus necator
SH
-
 Cupriavidus necator
SH i.e. soluble cytoplasmic hydrogenase Cupriavidus necator
soluble [NiFe]-hydrogenase
-
 Cupriavidus necator

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
30
-
 assay at Cupriavidus necator

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
8
-
 assay at Cupriavidus necator

Cofactor

Cofactor Comment Organism Structure
benzyl viologen oxidized and reduced Cupriavidus necator
FMN 2 molecules per enzyme molecule, one of 2 can be reversibly released upon reduction of the enzyme Cupriavidus necator
FMN required, 2 molecules per tetrameric enzyme complex, reduced enzyme reversibly releases half of the FMN bound, kinetics, FMN release induces reduction with NADH, overview Cupriavidus necator
NAD+
-
 Cupriavidus necator
NADH
-
 Cupriavidus necator