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Literature summary extracted from

  • Teufel, R.; Kung, J.; Kockelkorn, D.; Alber, B.; Fuchs, G.
    3-Hydroxypropionyl-coenzyme A dehydratase and acryloyl-coenzyme A reductase, enzymes of the autotrophic 3-hydroxypropionate/4-hydroxybutyrate cycle in the Sulfolobales (2009), J. Bacteriol., 191, 4572-4581.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.3.1.95 expressed in Escherichia coli Rosetta 2 (DE3) cells Sulfurisphaera tokodaii

Inhibitors

EC Number Inhibitors Comment Organism Structure
1.3.1.84 additional information not inhibitory: EDTA, Zn2+, other divalent cations Sulfurisphaera tokodaii

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.3.1.84 0.003
-
 acryloyl-CoA pH 6.0, 65°C Sulfurisphaera tokodaii
1.3.1.84 0.036
-
 NADPH pH 6.0, 65°C Sulfurisphaera tokodaii
1.3.1.95 0.01
-
 acryloyl-CoA apparent value, Km less than 0.01 mM, recombinant enzyme, at pH 7.5 and 22°C Sulfurisphaera tokodaii
1.3.1.95 0.01
-
 acryloyl-CoA apparent value, Km less than 0.01 mM, recombinant enzyme, at pH 7.5 and 22°C Metallosphaera sedula
1.3.1.95 0.036
-
 NADPH apparent value, recombinant enzyme, at pH 7.5 and 22°C Metallosphaera sedula
1.3.1.95 0.036
-
 acryloyl-CoA apparent value, recombinant enzyme, at pH 7.5 and 22°C Sulfurisphaera tokodaii
4.2.1.116 0.06
-
 3-hydroxypropionyl-CoA pH 8.6, 65°C Metallosphaera sedula
4.2.1.116 0.075
-
 (S)-3-hydroxybutyryl-CoA pH 8.6, 65°C Metallosphaera sedula

Localization

EC Number Localization Comment Organism GeneOntology No. Textmining
1.3.1.84 soluble
-
 Sulfurisphaera tokodaii
-
-

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
1.3.1.84 Zn2+ presence of 0.8 mol Zn2+ per mol of monomer, addition of Zn2+ is not stimulatory Sulfurisphaera tokodaii
1.3.1.95 Zn2+ contains 0.8 mol of Zn2+ per mol of enzyme monomer Sulfurisphaera tokodaii
1.3.1.95 Zn2+ contains 0.8 mol of Zn2+ per mol of enzyme monomer Metallosphaera sedula

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
1.3.1.84 38600
-
 1 * 39000, SDS-PAGE, 1 * 38600, calculated Sulfurisphaera tokodaii
1.3.1.84 39000
-
 1 * 39000, SDS-PAGE, 1 * 38600, calculated Sulfurisphaera tokodaii
1.3.1.84 40000
-
 x * 40000, SDS-PAGE Metallosphaera sedula
1.3.1.84 43000
-
 gel filtration Sulfurisphaera tokodaii
1.3.1.95 36000
-
 calculated from amino acid sequence Sulfurisphaera tokodaii
1.3.1.95 40000
-
 1 * 40000, SDS-PAGE Sulfurisphaera tokodaii
1.3.1.95 40000
-
 1 * 40000, SDS-PAGE Metallosphaera sedula
1.3.1.95 43000
-
 gel filtration Sulfurisphaera tokodaii
4.2.1.116 23000
-
 gel filtration Metallosphaera sedula
4.2.1.116 28300
-
 1 * 29000, SDS-PAGE, 1 * 28300, calculated Metallosphaera sedula
4.2.1.116 29000
-
 1 * 29000, SDS-PAGE, 1 * 28300, calculated Metallosphaera sedula

Organism

EC Number Organism UniProt Comment Textmining
1.3.1.84 Metallosphaera sedula
-
-
-
1.3.1.84 Sulfurisphaera tokodaii Q975C8
-
-
1.3.1.95 Metallosphaera sedula
-
-
-
1.3.1.95 Sulfurisphaera tokodaii
-
-
-
4.2.1.116 Metallosphaera sedula A4YI89
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.3.1.95 heat precipitation and Ni2+-chelating Sepharose column chromatography Sulfurisphaera tokodaii
1.3.1.95 heat precipitation, Q Sepharose column chromatography, carboxymethylcellulose column chromatography, ammonium sulfate precipitation, phenyl-Sepharose column chromatography, and Resource-S gel filtration Metallosphaera sedula

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
1.3.1.84 2.9
-
 65°C Metallosphaera sedula
1.3.1.84 18.7
-
 pH 6.0, 65°C Sulfurisphaera tokodaii
1.3.1.95 0.014
-
 cell extract, at pH 7.5 and 22°C Metallosphaera sedula
1.3.1.95 2.9
-
 after 210fold purification, at pH 7.5 and 22°C Metallosphaera sedula
1.3.1.95 4.6
-
 cell extract, at pH 7.5 and 22°C Sulfurisphaera tokodaii
1.3.1.95 18.7
-
 after 4.1fold purification, at pH 7.5 and 22°C Sulfurisphaera tokodaii
4.2.1.116 2.4
-
 extract of autotrophically grown cells, pH 8.6 Metallosphaera sedula
4.2.1.116 3.1
-
 extract of heterotrophically grown cells, pH 8.6 Metallosphaera sedula
4.2.1.116 151
-
 purified, recombinant protein, pH 8.6 Metallosphaera sedula

Storage Stability

EC Number Storage Stability Organism
1.3.1.84 -20°C, 30% glycerol, stable for months Sulfurisphaera tokodaii

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.3.1.84 acryloyl-CoA + NADPH + H+
-
 Sulfurisphaera tokodaii propanoyl-CoA + NADP+
-
 ?
1.3.1.84 acrylyl-CoA + NADPH + H+
-
 Metallosphaera sedula propanoyl-CoA + NADP+
-
 ?
1.3.1.84 additional information no substrate: NADH, crotonyl-CoA Sulfurisphaera tokodaii ?
-
 ?
1.3.1.95 acryloyl-CoA + NADPH + H+
-
 Metallosphaera sedula propanoyl-CoA + NADP+
-
 ir
1.3.1.95 acryloyl-CoA + NADPH + H+ 100% specificity Sulfurisphaera tokodaii propanoyl-CoA + NADP+
-
 ir
1.3.1.95 additional information less than 1% specificity for crotonyl-CoA Sulfurisphaera tokodaii ?
-
 ?
4.2.1.116 (S)-3-hydroxybutyryl-CoA no substrate: (R)-3-hydroxybutyryl-CoA Metallosphaera sedula crotonyl-CoA + H2O
-
 ?
4.2.1.116 3-hydroxypropanoyl-CoA
-
 Metallosphaera sedula acryloyl-CoA + H2O
-
 ?

Subunits

EC Number Subunits Comment Organism
1.3.1.84 ? x * 40000, SDS-PAGE Metallosphaera sedula
1.3.1.84 monomer 1 * 39000, SDS-PAGE, 1 * 38600, calculated Sulfurisphaera tokodaii
1.3.1.95 monomer 1 * 40000, SDS-PAGE Sulfurisphaera tokodaii
1.3.1.95 monomer 1 * 40000, SDS-PAGE Metallosphaera sedula
4.2.1.116 monomer 1 * 29000, SDS-PAGE, 1 * 28300, calculated Metallosphaera sedula

Synonyms

EC Number Synonyms Comment Organism
1.3.1.84 acryloyl-coenzyme A reductase
-
 Metallosphaera sedula
1.3.1.84 acryloyl-coenzyme A reductase
-
 Sulfurisphaera tokodaii
1.3.1.95 acryloyl-CoA reductase
-
 Sulfurisphaera tokodaii
1.3.1.95 acryloyl-CoA reductase
-
 Metallosphaera sedula
1.3.1.95 acryloyl-CoA reductase (NADPH)
-
 Sulfurisphaera tokodaii
1.3.1.95 acryloyl-CoA reductase (NADPH)
-
 Metallosphaera sedula
1.3.1.95 acryloyl-coenzyme A reductase
-
 Sulfurisphaera tokodaii
1.3.1.95 acryloyl-coenzyme A reductase
-
 Metallosphaera sedula

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.3.1.84 13
-
 NADPH pH 6.0, 65°C Sulfurisphaera tokodaii
1.3.1.95 13
-
 acryloyl-CoA apparent value, recombinant enzyme, at pH 7.5 and 22°C Sulfurisphaera tokodaii
1.3.1.95 13
-
 acryloyl-CoA apparent value, recombinant enzyme, at pH 7.5 and 22°C Metallosphaera sedula
4.2.1.116 96
-
 3-hydroxypropionyl-CoA pH 8.6, 65°C Metallosphaera sedula

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.3.1.84 6
-
-
 Sulfurisphaera tokodaii
1.3.1.95 6
-
-
 Sulfurisphaera tokodaii
4.2.1.116 8.1
-
-
 Metallosphaera sedula

pH Range

EC Number pH Minimum pH Maximum Comment Organism
1.3.1.84 7.5
-
 half-maximal activity Sulfurisphaera tokodaii
4.2.1.116 6.5
-
 half-maximal activity Metallosphaera sedula
4.2.1.116 9.5
-
 half-maximal activity Metallosphaera sedula

Cofactor

EC Number Cofactor Comment Organism Structure
1.3.1.84 NADPH no cofactor: NADH Sulfurisphaera tokodaii
1.3.1.95 FAD
-
 Sulfurisphaera tokodaii
1.3.1.95 FAD
-
 Metallosphaera sedula
1.3.1.95 NADPH
-
 Sulfurisphaera tokodaii
1.3.1.95 NADPH
-
 Metallosphaera sedula