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Literature summary 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

Cloned(Commentary)

EC Number Cloned (Comment) Organism
4.2.1.112 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)

EC Number Crystallization (Comment) Organism
4.2.1.112 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

EC Number Protein Variants Comment Organism
4.2.1.112 D13E site-directed mutagenesis, almost inactive mutant Syntrophotalea acetylenica
4.2.1.112 I142A site-directed mutagenesis, the mutant shows a marked loss of activity compared to the wild-type enzyme Syntrophotalea acetylenica
4.2.1.112 K48A site-directed mutagenesis, the mutant shows unaltered activity compared to the wild-type enzyme Syntrophotalea acetylenica
4.2.1.112 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]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
4.2.1.112 0.014
-
 Acetylene pH 7.5, 30°C Syntrophotalea acetylenica

Localization

EC Number Localization Comment Organism GeneOntology No. Textmining
4.2.1.112 soluble
-
 Syntrophotalea acetylenica
-
-

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
4.2.1.112 Iron acetylene hydratase is a tungsten, iron-sulfur enzyme, contains 4.8 mol of iron per mol of enzyme Syntrophotalea acetylenica
4.2.1.112 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)

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

Organism

EC Number Organism UniProt Comment Textmining
4.2.1.112 Syntrophotalea acetylenica
-
-
-
4.2.1.112 Syntrophotalea acetylenica WoAcy1 / DSM 3246
-
-
-

Reaction

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

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
4.2.1.112 additional information
-
 activity of different purifications, overview Syntrophotalea acetylenica
4.2.1.112 14.2
-
 purified recombinant enzyme, pH 7.5, 30°C Syntrophotalea acetylenica

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
4.2.1.112 acetylene + H2O
-
 Syntrophotalea acetylenica acetaldehyde
-
 r
4.2.1.112 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
4.2.1.112 acetylene + H2O
-
 Syntrophotalea acetylenica WoAcy1 / DSM 3246 acetaldehyde
-
 r
4.2.1.112 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

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

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
4.2.1.112 50
-
-
 Syntrophotalea acetylenica

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
4.2.1.112 6 6.5
-
 Syntrophotalea acetylenica

Cofactor

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