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Literature summary for 1.12.1.2 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)

Cloned (Comment) Organism
-
Cupriavidus necator

Protein Variants

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

Localization

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

Natural Substrates/ Products (Substrates)

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

Organism

Organism UniProt Comment Textmining
Cupriavidus necator
-
ATCC 17699, DSM 428
-

Reaction

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

Substrates and Products (Substrate)

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

Subunits

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

Synonyms

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

pH Optimum

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

Cofactor

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