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Literature summary extracted from

  • Ramos-Vera, W.H.; Weiss, M.; Strittmatter, E.; Kockelkorn, D.; Fuchs, G.
    Identification of missing genes and enzymes for autotrophic carbon fixation in crenarchaeota (2011), J. Bacteriol., 193, 1201-1211.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.1.1.35 expression in Escherichia coli Metallosphaera sedula
2.3.1.9 overexpressed as soluble N-terminal His10-tagged protein in Escherichia coli Pyrobaculum neutrophilum
4.2.1.150 expression in Escherichia coli Metallosphaera sedula
6.2.1.40 expression in Escherichia coli. The recombinant enzyme is modified by acetylation at Lys594 and is partly inactive Pyrobaculum neutrophilum

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.1.1.35 0.2
-
 (S)-3-hydroxybutanoyl-CoA pH 8.0, 65°C Metallosphaera sedula
1.1.1.35 0.25
-
 NAD+ pH 8.0, 65°C Metallosphaera sedula
2.3.1.9 0.15
-
 acetoacetyl-CoA pH 7.8, 85°C Pyrobaculum neutrophilum
4.2.1.150 0.3
-
 crotonyl-CoA pH 8.0, 65°C Metallosphaera sedula
6.2.1.40 0.7
-
 4-hydroxybutanoate pH 7.8, 85°C Pyrobaculum neutrophilum

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
1.1.1.35 70000
-
 x * 70000, SDS-PAGE Metallosphaera sedula
2.3.1.9 42444
-
 x * 42444, calculated from sequence Pyrobaculum neutrophilum
4.2.1.150 70000
-
 x * 70000, SDS-PAGE Metallosphaera sedula
6.2.1.40 71356
-
 x * 71356, calculated from sequence Pyrobaculum neutrophilum

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
1.1.1.35 (S)-3-hydroxybutanoyl-CoA + NAD+ Metallosphaera sedula the enzyme is involved in autotrophic carbon fixation acetoacetyl-CoA + NADH + H+
-
 ?
1.1.1.35 (S)-3-hydroxybutanoyl-CoA + NAD+ Metallosphaera sedula DSM 5348 the enzyme is involved in autotrophic carbon fixation acetoacetyl-CoA + NADH + H+
-
 ?
2.3.1.9 CoA + acetoacetyl-CoA Pyrobaculum neutrophilum the enzyme is involved in autotrophic carbon fixation 2 acetyl-CoA
-
 ?
2.3.1.9 CoA + acetoacetyl-CoA Pyrobaculum neutrophilum DSM 2338 the enzyme is involved in autotrophic carbon fixation 2 acetyl-CoA
-
 ?
4.2.1.150 crotonyl-CoA + H2O Metallosphaera sedula the enzyme is involved in autotrophic carbon fixation (S)-3-hydroxybutanoyl-CoA
-
 ?
4.2.1.150 crotonyl-CoA + H2O Metallosphaera sedula DSM 5348 the enzyme is involved in autotrophic carbon fixation (S)-3-hydroxybutanoyl-CoA
-
 ?
6.2.1.40 ATP + 4-hydroxybutanoate + CoA Pyrobaculum neutrophilum the enzyme is involved in autotrophic carbon fixation AMP + diphosphate + 4-hydroxybutanoyl-CoA
-
 ?
6.2.1.40 ATP + 4-hydroxybutanoate + CoA Pyrobaculum neutrophilum DSM 2338 the enzyme is involved in autotrophic carbon fixation AMP + diphosphate + 4-hydroxybutanoyl-CoA
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
1.1.1.35 Metallosphaera sedula A4YDS4
-
-
1.1.1.35 Metallosphaera sedula DSM 5348 A4YDS4
-
-
2.3.1.9 Pyrobaculum neutrophilum B1YB71
-
-
2.3.1.9 Pyrobaculum neutrophilum DSM 2338 B1YB71
-
-
4.2.1.150 Metallosphaera sedula A4YDS4
-
-
4.2.1.150 Metallosphaera sedula DSM 5348 A4YDS4
-
-
6.2.1.40 Pyrobaculum neutrophilum B1YBY4
-
-
6.2.1.40 Pyrobaculum neutrophilum DSM 2338 B1YBY4
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.1.1.35
-
 Metallosphaera sedula
2.3.1.9
-
 Pyrobaculum neutrophilum
4.2.1.150
-
 Metallosphaera sedula
6.2.1.40
-
 Pyrobaculum neutrophilum

Source Tissue

EC Number Source Tissue Comment Organism Textmining
1.1.1.35 culture condition:autotrophically-grown cell bifunctional crotonyl-CoA hydratase/(S)-3-hydroxybutanoyl-CoA dehydrogenase Metallosphaera sedula
-
1.1.1.35 culture condition:heterotrophically-grown cell bifunctional crotonyl-CoA hydratase/(S)-3-hydroxybutanoyl-CoA dehydrogenase Metallosphaera sedula
-
4.2.1.150 culture condition:autotrophically-grown cell
-
 Metallosphaera sedula
-
4.2.1.150 culture condition:heterotrophically-grown cell
-
 Metallosphaera sedula
-

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
1.1.1.35 3 8 pH 8.0, 65°C, native enzyme, autotrophically- or heterotrophically-grown cell Metallosphaera sedula
2.3.1.9 55
-
 pH 7.2, 85°C Pyrobaculum neutrophilum
4.2.1.150 3 8 pH 8.0, 65°C, native enzyme, autotrophically- or heterotrophically-grown cell Metallosphaera sedula
6.2.1.40 1.6
-
 pH 7.8, 85°C Pyrobaculum neutrophilum

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.1.1.35 (S)-3-hydroxybutanoyl-CoA + NAD+ the enzyme is involved in autotrophic carbon fixation Metallosphaera sedula acetoacetyl-CoA + NADH + H+
-
 ?
1.1.1.35 (S)-3-hydroxybutanoyl-CoA + NAD+ no activity with (R)-3-hydroxybutanoyl-CoA, no activity with NADP+. Bifunctional crotonyl-CoA hydratase/(S)-3-hydroxybutanoyl-CoA dehydrogenase (EC 4.2.1.150/EC 1.1.1.35) Metallosphaera sedula acetoacetyl-CoA + NADH + H+
-
 ?
1.1.1.35 (S)-3-hydroxybutanoyl-CoA + NAD+ the enzyme is involved in autotrophic carbon fixation Metallosphaera sedula DSM 5348 acetoacetyl-CoA + NADH + H+
-
 ?
1.1.1.35 (S)-3-hydroxybutanoyl-CoA + NAD+ no activity with (R)-3-hydroxybutanoyl-CoA, no activity with NADP+. Bifunctional crotonyl-CoA hydratase/(S)-3-hydroxybutanoyl-CoA dehydrogenase (EC 4.2.1.150/EC 1.1.1.35) Metallosphaera sedula DSM 5348 acetoacetyl-CoA + NADH + H+
-
 ?
2.3.1.9 CoA + acetoacetyl-CoA
-
 Pyrobaculum neutrophilum 2 acetyl-CoA
-
 ?
2.3.1.9 CoA + acetoacetyl-CoA the enzyme is involved in autotrophic carbon fixation Pyrobaculum neutrophilum 2 acetyl-CoA
-
 ?
2.3.1.9 CoA + acetoacetyl-CoA
-
 Pyrobaculum neutrophilum DSM 2338 2 acetyl-CoA
-
 ?
2.3.1.9 CoA + acetoacetyl-CoA the enzyme is involved in autotrophic carbon fixation Pyrobaculum neutrophilum DSM 2338 2 acetyl-CoA
-
 ?
4.2.1.150 crotonyl-CoA + H2O the enzyme is involved in autotrophic carbon fixation Metallosphaera sedula (S)-3-hydroxybutanoyl-CoA
-
 ?
4.2.1.150 crotonyl-CoA + H2O bifunctional crotonyl-CoA hydratase/(S)-3-hydroxybutanoyl-CoA dehydrogenase (EC 4.2.1.150/EC 1.1.1.35) Metallosphaera sedula (S)-3-hydroxybutanoyl-CoA
-
 ?
4.2.1.150 crotonyl-CoA + H2O the enzyme is involved in autotrophic carbon fixation Metallosphaera sedula DSM 5348 (S)-3-hydroxybutanoyl-CoA
-
 ?
4.2.1.150 crotonyl-CoA + H2O bifunctional crotonyl-CoA hydratase/(S)-3-hydroxybutanoyl-CoA dehydrogenase (EC 4.2.1.150/EC 1.1.1.35) Metallosphaera sedula DSM 5348 (S)-3-hydroxybutanoyl-CoA
-
 ?
6.2.1.40 ATP + 3-hydroxybutyrate + CoA 8% of the activity compared to 4-hydroxybutanoate Pyrobaculum neutrophilum AMP + diphosphate + 3-hydroxybutyryl-CoA
-
 ?
6.2.1.40 ATP + 3-hydroxybutyrate + CoA 8% of the activity compared to 4-hydroxybutanoate Pyrobaculum neutrophilum DSM 2338 AMP + diphosphate + 3-hydroxybutyryl-CoA
-
 ?
6.2.1.40 ATP + 3-hydroxypropionate + CoA 9% of the activity compared to 4-hydroxybutanoate Pyrobaculum neutrophilum AMP + diphosphate + 3-hydroxypropionyl-CoA
-
 ?
6.2.1.40 ATP + 4-hydroxybutanoate + CoA
-
 Pyrobaculum neutrophilum AMP + diphosphate + 4-hydroxybutanoyl-CoA
-
 ?
6.2.1.40 ATP + 4-hydroxybutanoate + CoA the enzyme is involved in autotrophic carbon fixation Pyrobaculum neutrophilum AMP + diphosphate + 4-hydroxybutanoyl-CoA
-
 ?
6.2.1.40 ATP + 4-hydroxybutanoate + CoA
-
 Pyrobaculum neutrophilum DSM 2338 AMP + diphosphate + 4-hydroxybutanoyl-CoA
-
 ?
6.2.1.40 ATP + 4-hydroxybutanoate + CoA the enzyme is involved in autotrophic carbon fixation Pyrobaculum neutrophilum DSM 2338 AMP + diphosphate + 4-hydroxybutanoyl-CoA
-
 ?
6.2.1.40 ATP + acetate + CoA 13% of the activity compared to 4-hydroxybutanoate Pyrobaculum neutrophilum AMP + diphosphate + acetyl-CoA
-
 ?
6.2.1.40 ATP + acetate + CoA 13% of the activity compared to 4-hydroxybutanoate Pyrobaculum neutrophilum DSM 2338 AMP + diphosphate + acetyl-CoA
-
 ?
6.2.1.40 ATP + crotonate + CoA 22% of the activity compared to 4-hydroxybutanoate Pyrobaculum neutrophilum AMP + diphosphate + crotonyl-CoA
-
 ?
6.2.1.40 ATP + crotonate + CoA 22% of the activity compared to 4-hydroxybutanoate Pyrobaculum neutrophilum DSM 2338 AMP + diphosphate + crotonyl-CoA
-
 ?

Subunits

EC Number Subunits Comment Organism
1.1.1.35 ? x * 70000, SDS-PAGE Metallosphaera sedula
2.3.1.9 ? x * 42444, calculated from sequence Pyrobaculum neutrophilum
4.2.1.150 ? x * 70000, SDS-PAGE Metallosphaera sedula
6.2.1.40 ? x * 71356, calculated from sequence Pyrobaculum neutrophilum

Synonyms

EC Number Synonyms Comment Organism
1.1.1.35 Msed_0399 locus name, bifunctional crotonyl-CoA hydratase/(S)-3-hydroxybutanoyl-CoA dehydrogenase (EC 4.2.1.150/EC 1.1.1.35) Metallosphaera sedula
2.3.1.9 beta-ketothiolase
-
 Pyrobaculum neutrophilum
2.3.1.9 Tneu_0249 locus name Pyrobaculum neutrophilum
4.2.1.150 Msed_0399 locus name, bifunctional crotonyl-CoA hydratase/(S)-3-hydroxybutanoyl-CoA dehydrogenase (EC 4.2.1.150/EC 1.1.1.35) Metallosphaera sedula
6.2.1.40 Tneu_0420 locus name Pyrobaculum neutrophilum

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.1.1.35 65
-
 assay at Metallosphaera sedula
4.2.1.150 65
-
 assay at Metallosphaera sedula
6.2.1.40 85
-
 assay at Pyrobaculum neutrophilum

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.1.1.35 8
-
 assay at Metallosphaera sedula
4.2.1.150 8
-
 assay at Metallosphaera sedula
6.2.1.40 7.8
-
 assay at Pyrobaculum neutrophilum

Cofactor

EC Number Cofactor Comment Organism Structure
1.1.1.35 NAD+ no activity with NADP+ Metallosphaera sedula

General Information

EC Number General Information Comment Organism
1.1.1.35 physiological function the enzyme is involved in autotrophic carbon fixation Metallosphaera sedula
2.3.1.9 physiological function the enzyme is involved in autotrophic carbon fixation Pyrobaculum neutrophilum
4.2.1.150 physiological function the enzyme is involved in autotrophic carbon fixation Metallosphaera sedula
6.2.1.40 physiological function the enzyme is involved in autotrophic carbon fixation Pyrobaculum neutrophilum