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Literature summary extracted from

  • Burgdorf, T.; De Lacey, A.L.; Friedrich, B.
    Functional analysis by site-directed mutagenesis of the NAD+-reducing hydrogenase from Ralstonia eutropha (2002), J. Bacteriol., 184, 6280-6288.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.12.1.2
-
Cupriavidus necator
1.12.99.6
-
Cupriavidus necator

Protein Variants

EC Number Protein Variants Comment Organism
1.12.1.2 C461A site-directed mutagenesis, catalytic subunit HoxH allele, mutant strain does not grow, inactive Cupriavidus necator
1.12.1.2 D456N site-directed mutagenesis, catalytic subunit HoxH allele, mutant strain does not grow, nearly inactive Cupriavidus necator
1.12.1.2 D456S site-directed mutagenesis, catalytic subunit HoxH allele, mutant strain does not grow, nearly inactive Cupriavidus necator
1.12.1.2 E14Q site-directed mutagenesis, catalytic subunit HoxH allele, mutant strain does not grow, inactive Cupriavidus necator
1.12.1.2 E14V site-directed mutagenesis, catalytic subunit HoxH allele, mutant strain does not grow, nearly inactive Cupriavidus necator
1.12.1.2 G15A site-directed mutagenesis, catalytic subunit HoxH allele, mutant strain does not grow, nearly inactive Cupriavidus necator
1.12.1.2 H16L site-directed mutagenesis, catalytic subunit HoxH allele, mutant strain does not grow, highly reduced activity compared to the wild-type strain Cupriavidus necator
1.12.1.2 H396L site-directed mutagenesis, catalytic subunit HoxH allele, highly reduced activity compared to the wild-type strain Cupriavidus necator
1.12.1.2 H396Q site-directed mutagenesis, catalytic subunit HoxH allele, about 3.5fold reduced activity compared to the wild-type strain Cupriavidus necator
1.12.1.2 H69Q site-directed mutagenesis, catalytic subunit HoxH allele, about 5fold reduced activity compared to the wild-type strain Cupriavidus necator
1.12.1.2 H69Q/P390A site-directed mutagenesis, catalytic subunit HoxH alleles, reduced activity compared to the wild-type strain Cupriavidus necator
1.12.1.2 I64A site-directed mutagenesis, catalytic subunit HoxH allele, mutant strain does not grow, highly reduced activity compared to the wild-type strain Cupriavidus necator
1.12.1.2 L118A site-directed mutagenesis, catalytic subunit HoxH allele, reduced activity compared to the wild-type strain Cupriavidus necator
1.12.1.2 L118F site-directed mutagenesis, catalytic subunit HoxH allele, mutant strain does not grow at O2 concentrations above 5%, about 4fold reduced activity compared to the wild-type strain Cupriavidus necator
1.12.1.2 L118I site-directed mutagenesis, catalytic subunit HoxH allele, reduced activity compared to the wild-type strain Cupriavidus necator
1.12.1.2 L394A site-directed mutagenesis, catalytic subunit HoxH allele, about 5fold reduced activity compared to the wild-type strain Cupriavidus necator
1.12.1.2 L394N site-directed mutagenesis, catalytic subunit HoxH allele, mutant strain does not grow, nearly inactive Cupriavidus necator
1.12.1.2 additional information construction of an inactive null mutant strain, mutant strain growth characteristics, overview Cupriavidus necator
1.12.1.2 P390A site-directed mutagenesis, catalytic subunit HoxH allele, about 5fold reduced activity compared to the wild-type strain Cupriavidus necator
1.12.1.2 R12L site-directed mutagenesis, catalytic subunit HoxH allele, mutant strain does not grow, highly reduced activity compared to the wild-type strain Cupriavidus necator
1.12.1.2 R12Q site-directed mutagenesis, catalytic subunit HoxH allele, mutant strain does not grow, highly reduced activity compared to the wild-type strain Cupriavidus necator
1.12.1.2 R391L site-directed mutagenesis, catalytic subunit HoxH allele, mutant strain does not grow, inactive Cupriavidus necator
1.12.1.2 R391Q site-directed mutagenesis, catalytic subunit HoxH allele, mutant strain does not grow, inactive Cupriavidus necator
1.12.1.2 R60Q site-directed mutagenesis, catalytic subunit HoxH allele, highly reduced activity compared to the wild-type strain Cupriavidus necator
1.12.1.2 S68V site-directed mutagenesis, catalytic subunit HoxH allele, highly reduced activity compared to the wild-type strain Cupriavidus necator
1.12.1.2 T415S site-directed mutagenesis, catalytic subunit HoxH allele, about 5fold reduced activity compared to the wild-type strain Cupriavidus necator
1.12.1.2 T415V site-directed mutagenesis, catalytic subunit HoxH allele, highly reduced activity compared to the wild-type strain Cupriavidus necator
1.12.1.2 T415V/N415H site-directed mutagenesis, catalytic subunit HoxH alleles, O2-sensitive growth, highly reduced activity compared to the wild-type strain Cupriavidus necator
1.12.99.6 additional information site directed mutagenesis in conserved motifs of the subunit HoxH Cupriavidus necator

Localization

EC Number Localization Comment Organism GeneOntology No. Textmining
1.12.1.2 cytosol
-
Cupriavidus necator 5829
-

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
1.12.1.2 H2 + NAD+ Cupriavidus necator
-
H+ + NADH
-
?
1.12.1.2 additional information Cupriavidus necator activity is equally high in atmosperes of 20% O2, 20% N2, or 100% H2, the latter being slightly preferred ?
-
?

Organism

EC Number Organism UniProt Comment Textmining
1.12.1.2 Cupriavidus necator
-
ATCC 17699, DSM 428
-
1.12.99.6 Cupriavidus necator
-
wild type and mutants
-

Reaction

EC Number Reaction Comment Organism Reaction ID
1.12.1.2 H2 + NAD+ = H+ + NADH molecular mechanism of hydrogen activation Cupriavidus necator

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.12.1.2 H2 + NAD+
-
Cupriavidus necator H+ + NADH
-
?
1.12.1.2 H2 + oxidized benzyl viologen
-
Cupriavidus necator H+ + reduced benzyl viologen
-
?
1.12.1.2 additional information activity is equally high in atmosperes of 20% O2, 20% N2, or 100% H2, the latter being slightly preferred Cupriavidus necator ?
-
?
1.12.1.2 additional information deuterium-hydrogen proton exchange activity of wild-type and mutantenzymes, overview Cupriavidus necator ?
-
?
1.12.99.6 H2 + acceptor
-
Cupriavidus necator H+ + reduced acceptor
-
?

Subunits

EC Number Subunits Comment Organism
1.12.1.2 tetramer the catalytic subunit is termed HoxH Cupriavidus necator

Synonyms

EC Number Synonyms Comment Organism
1.12.1.2 NAD+-reducing hydrogenase
-
Cupriavidus necator
1.12.1.2 SH
-
Cupriavidus necator
1.12.1.2 soluble hydrogenase
-
Cupriavidus necator
1.12.1.2 [NiFe] hydrogenase
-
Cupriavidus necator

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.12.1.2 7
-
assay at Cupriavidus necator

Cofactor

EC Number Cofactor Comment Organism Structure
1.12.1.2 benzyl viologen oxidized and reduced Cupriavidus necator
1.12.1.2 NAD+
-
Cupriavidus necator
1.12.1.2 NADH
-
Cupriavidus necator