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Literature summary extracted from

  • Yoon, K.S.; Sakai, Y.; Tsukada, N.; Fujisawa, K.; Nishihara, H.
    Purification and biochemical characterization of a membrane-bound [NiFe]-hydrogenase from a hydrogen-oxidizing, lithotrophic bacterium, Hydrogenophaga sp. AH-24 (2009), FEMS Microbiol. Lett., 290, 114-120.
    View publication on PubMed

Activating Compound

EC Number Activating Compound Comment Organism Structure
1.12.99.6 additional information activity increases up to 2.5fold during incubation at 50°C under H2 Hydrogenophaga sp.

Inhibitors

EC Number Inhibitors Comment Organism Structure
1.12.99.6 O2 enzyme is quite stable under air atmosphere, half-life of activity is ca. 48 h at 4°C, inactivation is irreversible Hydrogenophaga sp.

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.12.99.6 0.025
-
 oxidized methylene blue pH 7.0, 30°C Hydrogenophaga sp.
1.12.99.6 1.78
-
 benzyl viologen pH 7.0, 30°C Hydrogenophaga sp.
1.12.99.6 12.5
-
 methyl viologen pH 7.0, 30°C Hydrogenophaga sp.

Localization

EC Number Localization Comment Organism GeneOntology No. Textmining
1.12.99.6 membrane
-
 Hydrogenophaga sp. 16020
-

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
1.12.99.6 Fe membrane-bound [NiFe]-hydrogenase exhibits prominent electron paramagnetic resonance signals originating from [3Fe–4S]1 and [4Fe–4S]1 clusters Hydrogenophaga sp.

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
1.12.99.6 37000
-
 1 * 62000 + 1 * 37000, SDS-PAGE Hydrogenophaga sp.
1.12.99.6 62000
-
 1 * 62000 + 1 * 37000, SDS-PAGE Hydrogenophaga sp.
1.12.99.6 100000
-
 gel filtration Hydrogenophaga sp.

Organism

EC Number Organism UniProt Comment Textmining
1.12.99.6 Hydrogenophaga sp.
-
 AH-24
-
1.12.99.6 Hydrogenophaga sp. AH-24
-
 AH-24
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.12.99.6 to homogeneity, by gel filtration, 133fold purified and 68.2% of yield relative to the membrane Hydrogenophaga sp.

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
1.12.99.6 110
-
 purified enzyme Hydrogenophaga sp.

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.12.99.6 H2 + oxidized benzyl viologen
-
 Hydrogenophaga sp. H+ + reduced benzyl viologen
-
 ?
1.12.99.6 H2 + oxidized benzyl viologen
-
 Hydrogenophaga sp. AH-24 H+ + reduced benzyl viologen
-
 ?
1.12.99.6 H2 + oxidized methyl viologen
-
 Hydrogenophaga sp. H+ + reduced methyl viologen
-
 ?
1.12.99.6 H2 + oxidized methyl viologen
-
 Hydrogenophaga sp. AH-24 H+ + reduced methyl viologen
-
 ?
1.12.99.6 H2 + oxidized methylene blue
-
 Hydrogenophaga sp. H+ + reduced methylene blue
-
 ?
1.12.99.6 H2 + oxidized methylene blue
-
 Hydrogenophaga sp. AH-24 H+ + reduced methylene blue
-
 ?
1.12.99.6 H2 + oxidized methylene blue
-
 Hydrogenophaga sp. reduced methylene blue + H+
-
 r
1.12.99.6 H2 + oxidized methylene blue
-
 Hydrogenophaga sp. AH-24 reduced methylene blue + H+
-
 r
1.12.99.6 H2 + oxidized methylene blue
-
 Hydrogenophaga sp. reduced methylene blue
-
 r
1.12.99.6 H2 + oxidized methylene blue
-
 Hydrogenophaga sp. AH-24 reduced methylene blue
-
 r

Subunits

EC Number Subunits Comment Organism
1.12.99.6 heterodimer 1 * 62000 + 1 * 37000, SDS-PAGE Hydrogenophaga sp.

Synonyms

EC Number Synonyms Comment Organism
1.12.99.6 Mbh
-
 Hydrogenophaga sp.
1.12.99.6 membrane-bound [NiFe]-hydrogenase
-
 Hydrogenophaga sp.

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.12.99.6 75
-
-
 Hydrogenophaga sp.

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
1.12.99.6 50
-
 under anaerobic conditions, retains full activity for over 5 h Hydrogenophaga sp.

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.12.99.6 20
-
 methyl viologen pH 7.0, 30°C Hydrogenophaga sp.
1.12.99.6 240
-
 benzyl viologen pH 7.0, 30°C Hydrogenophaga sp.
1.12.99.6 560
-
 methylene blue pH 7.0, 30°C Hydrogenophaga sp.

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.12.99.6 4
-
 H2 evolution Hydrogenophaga sp.
1.12.99.6 8
-
 H2 oxidation Hydrogenophaga sp.

pH Range

EC Number pH Minimum pH Maximum Comment Organism
1.12.99.6 3.5 9 H2 evolution, pH 3.5 and 9.0: ca. 50% of maximal activity Hydrogenophaga sp.
1.12.99.6 5.5 9 H2 oxidation, pH 5.5: 40% of maximal activity, pH 9: 60% of maximal activity Hydrogenophaga sp.

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
1.12.99.6 1.6
-
 methyl viologen pH 7.0, 30°C Hydrogenophaga sp.
1.12.99.6 135
-
 benzyl viologen pH 7.0, 30°C Hydrogenophaga sp.
1.12.99.6 22400
-
 methylene blue pH 7.0, 30°C Hydrogenophaga sp.