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Literature summary for 4.2.1.112 extracted from

  • Tenbrink, F.; Schink, B.; Kroneck, P.M.
    Exploring the active site of the tungsten, iron-sulfur enzyme acetylene hydratase (2011), J. Bacteriol., 193, 1229-1236.
    View publication on PubMedView publication on EuropePMC
Search for more literature for 4.2.1.112

Cloned(Commentary)

Cloned (Comment) Organism
cloning of AH gene in Escherichia coli strain JM109, expression of wild-type enzyme, active-site variants of the enzyme, and of the nitrate reductase N-terminal chaperone binding site NarG-fusion enzyme in Escherichia coli strain BL21(DE3) Syntrophotalea acetylenica

Crystallization (Commentary)

Crystallization (Comment) Organism
purified recombinant NarG-fusion acetylene hydratase, sitting and hanging drop vapor diffusion methods, small crystals after 3 to 4 weeks, 6.5-10 mg/ml protein in solution containing 5 mM HEPES-NaOH, pH 7.5, and 7.5 mM Na2S2O4, cryoprotection by 20% v/v 2-methyl-2,4-pentanediol, X-ray diffraction structure determination and analysis Syntrophotalea acetylenica

Protein Variants

Protein Variants Comment Organism
D13E site-directed mutagenesis, almost inactive mutant Syntrophotalea acetylenica
I142A site-directed mutagenesis, the mutant shows a marked loss of activity compared to the wild-type enzyme Syntrophotalea acetylenica
K48A site-directed mutagenesis, the mutant shows unaltered activity compared to the wild-type enzyme Syntrophotalea acetylenica
additional information construction of active-site variants, and of a fusion protein of the N-terminal chaperone binding site of the Escherichia coli nitrate reductase NarG to the AH gene improving the yield and activity of AH and its variants significantly, overview Syntrophotalea acetylenica

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.014
-
 Acetylene pH 7.5, 30°C Syntrophotalea acetylenica

Localization

Localization Comment Organism GeneOntology No. Textmining
soluble
-
 Syntrophotalea acetylenica
-
-

Metals/Ions

Metals/Ions Comment Organism Structure
Iron acetylene hydratase is a tungsten, iron-sulfur enzyme, contains 4.8 mol of iron per mol of enzyme Syntrophotalea acetylenica
Tungsten acetylene hydratase is a tungsten, iron-sulfur enzyme, active W(IV) state structure, contains 0.4 mol of tungsten per mol of enzymeoverview Syntrophotalea acetylenica

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
acetylene + H2O Syntrophotalea acetylenica
-
 acetaldehyde
-
 r
acetylene + H2O Syntrophotalea acetylenica WoAcy1 / DSM 3246
-
 acetaldehyde
-
 r

Organism

Organism UniProt Comment Textmining
Syntrophotalea acetylenica
-
-
-
Syntrophotalea acetylenica WoAcy1 / DSM 3246
-
-
-

Reaction

Reaction Comment Organism Reaction ID
acetaldehyde = acetylene + H2O active-site access, active-site architecture, and reaction mechanism, overview Syntrophotalea acetylenica
a

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
additional information
-
 activity of different purifications, overview Syntrophotalea acetylenica
14.2
-
 purified recombinant enzyme, pH 7.5, 30°C Syntrophotalea acetylenica

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
acetylene + H2O
-
 Syntrophotalea acetylenica acetaldehyde
-
 r
acetylene + H2O the active site residues Asp13, Lys48, and Ile142 are involved in catalysis. Asp13 close to W(IV) coordinates two molybdopterin-guanosine-dinucleotide ligands, Lys48 couples the [4Fe-4S] cluster to the W site, and Ile142 is part of a hydrophobic ring at the end of the substrate access channel designed to accommodate the substrate acetylene Syntrophotalea acetylenica acetaldehyde
-
 r
acetylene + H2O
-
 Syntrophotalea acetylenica WoAcy1 / DSM 3246 acetaldehyde
-
 r
acetylene + H2O the active site residues Asp13, Lys48, and Ile142 are involved in catalysis. Asp13 close to W(IV) coordinates two molybdopterin-guanosine-dinucleotide ligands, Lys48 couples the [4Fe-4S] cluster to the W site, and Ile142 is part of a hydrophobic ring at the end of the substrate access channel designed to accommodate the substrate acetylene Syntrophotalea acetylenica WoAcy1 / DSM 3246 acetaldehyde
-
 r

Subunits

Subunits Comment Organism
More analysis of the active W(IV) state three-dimensional structure, overview Syntrophotalea acetylenica

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
50
-
-
 Syntrophotalea acetylenica

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
6 6.5
-
 Syntrophotalea acetylenica

Cofactor

Cofactor Comment Organism Structure
molybdenum cofactor molybdopterin cofactor, 0.94 mol per mol of enzyme in purified recombinant enzyme Syntrophotalea acetylenica