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Literature summary extracted from

  • Hawkins, A.B.; Adams, M.W.; Kelly, R.M.
    Conversion of 4-hydroxybutyrate to acetyl coenzyme A and its anapleurosis in the Metallosphaera sedula 3-hydroxypropionate/4-hydroxybutyrate carbon fixation pathway (2014), Appl. Environ. Microbiol., 80, 2536-2545.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.1.1.35 recombinantly expressed in Escherichia coli Metallosphaera sedula
2.3.1.9 recombinantly expressed in Escherichia coli Metallosphaera sedula
4.2.1.17 expressed in Escherichia coli Rosetta 2 (DE3) cells Metallosphaera sedula
4.2.1.120 recombinantly expressed in Escherichia coli Metallosphaera sedula
4.2.1.150 recombinantly expressed in Escherichia coli Metallosphaera sedula
6.2.1.40 recombinantly expressed in Escherichia coli Metallosphaera sedula

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.1.1.35 0.06
-
 (S)-3-hydroxybutanoyl-CoA pH 8.0, 70°C Metallosphaera sedula
2.3.1.9 0.18
-
 acetoacetyl-CoA pH 7.5, 70°C Metallosphaera sedula
4.2.1.17 0.06
-
 (S)-3-hydroxybutyryl-CoA at pH 7.0 and 70°C Metallosphaera sedula
4.2.1.17 0.07
-
 crotonoyl-CoA at pH 7.0 and 70°C Metallosphaera sedula
4.2.1.120 0.15
-
 4-hydroxybutanoyl-CoA pH 8.0, 70°C Metallosphaera sedula
4.2.1.150 0.07
-
 crotonyl-CoA pH 8.0, 70°C Metallosphaera sedula
6.2.1.40 2
-
 4-hydroxybutanoate pH 7.5, 70°C Metallosphaera sedula

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 the 3-hydroxypropionate/4-hydroxybutyrate carbon fixation pathway acetoacetyl-CoA + NADH + H+
-
 ?
1.1.1.35 (S)-3-hydroxybutanoyl-CoA + NAD+ Metallosphaera sedula DSM 5348 the enzyme is involved in the 3-hydroxypropionate/4-hydroxybutyrate carbon fixation pathway acetoacetyl-CoA + NADH + H+
-
 ?
2.3.1.9 CoA + acetoacetyl-CoA Metallosphaera sedula the enzyme is involved in the 3-hydroxypropionate/4-hydroxybutyrate carbon fixation pathway 2 acetyl-CoA
-
 ?
2.3.1.9 CoA + acetoacetyl-CoA Metallosphaera sedula DSM 5348 the enzyme is involved in the 3-hydroxypropionate/4-hydroxybutyrate carbon fixation pathway 2 acetyl-CoA
-
 ?
4.2.1.17 (S)-3-hydroxybutyryl-CoA Metallosphaera sedula
-
 crotonoyl-CoA + H2O
-
 r
4.2.1.17 (S)-3-hydroxybutyryl-CoA Metallosphaera sedula DSM 5348
-
 crotonoyl-CoA + H2O
-
 r
4.2.1.17 crotonoyl-CoA + H2O Metallosphaera sedula
-
 (S)-3-hydroxybutyryl-CoA
-
 r
4.2.1.17 crotonoyl-CoA + H2O Metallosphaera sedula DSM 5348
-
 (S)-3-hydroxybutyryl-CoA
-
 r
4.2.1.120 4-hydroxybutanoyl-CoA Metallosphaera sedula the enzyme is involved in the 3-hydroxypropionate/4-hydroxybutyrate carbon fixation pathway crotonyl-CoA + H2O
-
 ?
4.2.1.120 4-hydroxybutanoyl-CoA Metallosphaera sedula DSM 5348 the enzyme is involved in the 3-hydroxypropionate/4-hydroxybutyrate carbon fixation pathway crotonyl-CoA + H2O
-
 ?
4.2.1.150 crotonyl-CoA + H2O Metallosphaera sedula the enzyme is involved in the 3-hydroxypropionate/4-hydroxybutyrate carbon fixation pathway (S)-3-hydroxybutanoyl-CoA
-
 ?
4.2.1.150 crotonyl-CoA + H2O Metallosphaera sedula DSM 5348 the enzyme is involved in the 3-hydroxypropionate/4-hydroxybutyrate carbon fixation pathway (S)-3-hydroxybutanoyl-CoA
-
 ?
6.2.1.40 ATP + 4-hydroxybutanoate + CoA Metallosphaera sedula the enzyme is involved in the 3-hydroxypropionate/4-hydroxybutyrate carbon fixation pathway. The flux between the succinate and acetyl-CoA branches in the 3-hydroxypropionate/4-hydroxybutyrate carbon fixation pathway is governed by 4-hydroxybutyrate-CoA ligase, possibly regulated posttranslationally by the protein acetyltransferase AMP + diphosphate + 4-hydroxybutanoyl-CoA
-
 ?
6.2.1.40 ATP + 4-hydroxybutanoate + CoA Metallosphaera sedula DSM 5348 the enzyme is involved in the 3-hydroxypropionate/4-hydroxybutyrate carbon fixation pathway. The flux between the succinate and acetyl-CoA branches in the 3-hydroxypropionate/4-hydroxybutyrate carbon fixation pathway is governed by 4-hydroxybutyrate-CoA ligase, possibly regulated posttranslationally by the protein acetyltransferase 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
-
-
1.1.1.157 Metallosphaera sedula A4YDS4
-
-
1.1.1.157 Metallosphaera sedula DSM 5348 A4YDS4
-
-
2.3.1.9 Metallosphaera sedula A4YEH9
-
-
2.3.1.9 Metallosphaera sedula DSM 5348 A4YEH9
-
-
4.2.1.17 Metallosphaera sedula
-
-
-
4.2.1.17 Metallosphaera sedula DSM 5348
-
-
-
4.2.1.120 Metallosphaera sedula A4YGC7
-
-
4.2.1.120 Metallosphaera sedula DSM 5348 A4YGC7
-
-
4.2.1.150 Metallosphaera sedula A4YDS4
-
-
4.2.1.150 Metallosphaera sedula DSM 5348 A4YDS4
-
-
6.2.1.40 Metallosphaera sedula A4YDT1
-
-
6.2.1.40 Metallosphaera sedula DSM 5348 A4YDT1
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
4.2.1.17 Q-Sepharose column chromatography and Superdex 200 gel filtration Metallosphaera sedula

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.1.1.35 (S)-3-hydroxybutanoyl-CoA + NAD+
-
 Metallosphaera sedula acetoacetyl-CoA + NADH + H+
-
 ?
1.1.1.35 (S)-3-hydroxybutanoyl-CoA + NAD+ the enzyme is involved in the 3-hydroxypropionate/4-hydroxybutyrate carbon fixation pathway Metallosphaera sedula acetoacetyl-CoA + NADH + H+
-
 ?
1.1.1.35 (S)-3-hydroxybutanoyl-CoA + NAD+
-
 Metallosphaera sedula DSM 5348 acetoacetyl-CoA + NADH + H+
-
 ?
1.1.1.35 (S)-3-hydroxybutanoyl-CoA + NAD+ the enzyme is involved in the 3-hydroxypropionate/4-hydroxybutyrate carbon fixation pathway Metallosphaera sedula DSM 5348 acetoacetyl-CoA + NADH + H+
-
 ?
2.3.1.9 CoA + acetoacetyl-CoA
-
 Metallosphaera sedula 2 acetyl-CoA
-
 ?
2.3.1.9 CoA + acetoacetyl-CoA the enzyme is involved in the 3-hydroxypropionate/4-hydroxybutyrate carbon fixation pathway Metallosphaera sedula 2 acetyl-CoA
-
 ?
2.3.1.9 CoA + acetoacetyl-CoA
-
 Metallosphaera sedula DSM 5348 2 acetyl-CoA
-
 ?
2.3.1.9 CoA + acetoacetyl-CoA the enzyme is involved in the 3-hydroxypropionate/4-hydroxybutyrate carbon fixation pathway Metallosphaera sedula DSM 5348 2 acetyl-CoA
-
 ?
4.2.1.17 (S)-3-hydroxybutyryl-CoA
-
 Metallosphaera sedula crotonoyl-CoA + H2O
-
 r
4.2.1.17 (S)-3-hydroxybutyryl-CoA
-
 Metallosphaera sedula DSM 5348 crotonoyl-CoA + H2O
-
 r
4.2.1.17 crotonoyl-CoA + H2O
-
 Metallosphaera sedula (S)-3-hydroxybutyryl-CoA
-
 r
4.2.1.17 crotonoyl-CoA + H2O
-
 Metallosphaera sedula DSM 5348 (S)-3-hydroxybutyryl-CoA
-
 r
4.2.1.120 4-hydroxybutanoyl-CoA
-
 Metallosphaera sedula crotonyl-CoA + H2O
-
 ?
4.2.1.120 4-hydroxybutanoyl-CoA the enzyme is involved in the 3-hydroxypropionate/4-hydroxybutyrate carbon fixation pathway Metallosphaera sedula crotonyl-CoA + H2O
-
 ?
4.2.1.120 4-hydroxybutanoyl-CoA
-
 Metallosphaera sedula DSM 5348 crotonyl-CoA + H2O
-
 ?
4.2.1.120 4-hydroxybutanoyl-CoA the enzyme is involved in the 3-hydroxypropionate/4-hydroxybutyrate carbon fixation pathway Metallosphaera sedula DSM 5348 crotonyl-CoA + H2O
-
 ?
4.2.1.150 crotonyl-CoA + H2O
-
 Metallosphaera sedula (S)-3-hydroxybutanoyl-CoA
-
 ?
4.2.1.150 crotonyl-CoA + H2O the enzyme is involved in the 3-hydroxypropionate/4-hydroxybutyrate carbon fixation pathway Metallosphaera sedula (S)-3-hydroxybutanoyl-CoA
-
 ?
4.2.1.150 crotonyl-CoA + H2O
-
 Metallosphaera sedula DSM 5348 (S)-3-hydroxybutanoyl-CoA
-
 ?
4.2.1.150 crotonyl-CoA + H2O the enzyme is involved in the 3-hydroxypropionate/4-hydroxybutyrate carbon fixation pathway Metallosphaera sedula DSM 5348 (S)-3-hydroxybutanoyl-CoA
-
 ?
6.2.1.40 ATP + 4-hydroxybutanoate + CoA
-
 Metallosphaera sedula AMP + diphosphate + 4-hydroxybutanoyl-CoA
-
 ?
6.2.1.40 ATP + 4-hydroxybutanoate + CoA the enzyme is involved in the 3-hydroxypropionate/4-hydroxybutyrate carbon fixation pathway. The flux between the succinate and acetyl-CoA branches in the 3-hydroxypropionate/4-hydroxybutyrate carbon fixation pathway is governed by 4-hydroxybutyrate-CoA ligase, possibly regulated posttranslationally by the protein acetyltransferase Metallosphaera sedula AMP + diphosphate + 4-hydroxybutanoyl-CoA
-
 ?
6.2.1.40 ATP + 4-hydroxybutanoate + CoA
-
 Metallosphaera sedula DSM 5348 AMP + diphosphate + 4-hydroxybutanoyl-CoA
-
 ?
6.2.1.40 ATP + 4-hydroxybutanoate + CoA the enzyme is involved in the 3-hydroxypropionate/4-hydroxybutyrate carbon fixation pathway. The flux between the succinate and acetyl-CoA branches in the 3-hydroxypropionate/4-hydroxybutyrate carbon fixation pathway is governed by 4-hydroxybutyrate-CoA ligase, possibly regulated posttranslationally by the protein acetyltransferase Metallosphaera sedula DSM 5348 AMP + diphosphate + 4-hydroxybutanoyl-CoA
-
 ?

Synonyms

EC Number Synonyms Comment Organism
1.1.1.35 (S)-3-hydroxybutyryl-CoA dehydrogenase
-
 Metallosphaera sedula
1.1.1.35 Msed_0399 locus name Metallosphaera sedula
1.1.1.157 (S)-3-hydroxybutyryl-CoA dehydrogenase
-
 Metallosphaera sedula
1.1.1.157 Msed_0399 locus name Metallosphaera sedula
2.3.1.9 Msed_0656 locus name Metallosphaera sedula
4.2.1.17 CCH/HBCD
-
 Metallosphaera sedula
4.2.1.17 crotonyl-CoA hydratase
-
 Metallosphaera sedula
4.2.1.120 Msed_1321 locus name Metallosphaera sedula
4.2.1.150 crotonyl-CoA hydratase
-
 Metallosphaera sedula
4.2.1.150 Msed_0399 locus name Metallosphaera sedula
6.2.1.40 4-hydroxybutyrate-CoA ligase (AMP forming)
-
 Metallosphaera sedula
6.2.1.40 Msed_0406 locus name Metallosphaera sedula

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.1.1.35 70
-
 assay at Metallosphaera sedula
2.3.1.9 70
-
 assay at Metallosphaera sedula
4.2.1.120 70
-
 assay at Metallosphaera sedula
4.2.1.150 70
-
 assay at Metallosphaera sedula
6.2.1.40 70
-
 assay at Metallosphaera sedula

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.1.1.35 15
-
 (S)-3-hydroxybutanoyl-CoA pH 8.0, 70°C Metallosphaera sedula
2.3.1.9 1000
-
 acetoacetyl-CoA pH 7.5, 70°C Metallosphaera sedula
4.2.1.17 15
-
 (S)-3-hydroxybutyryl-CoA at pH 7.0 and 70°C Metallosphaera sedula
4.2.1.17 19
-
 crotonoyl-CoA at pH 7.0 and 70°C Metallosphaera sedula
4.2.1.120 2.1
-
 4-hydroxybutanoyl-CoA pH 8.0, 70°C Metallosphaera sedula
4.2.1.150 19
-
 crotonyl-CoA pH 8.0, 70°C Metallosphaera sedula
6.2.1.40 1.8
-
 4-hydroxybutanoate pH 7.5, 70°C Metallosphaera sedula

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.1.1.35 8
-
 assay at Metallosphaera sedula
2.3.1.9 7.5
-
 assay at Metallosphaera sedula
4.2.1.120 8
-
 assay at Metallosphaera sedula
4.2.1.150 8
-
 assay at Metallosphaera sedula
6.2.1.40 7.5
-
 assay at Metallosphaera sedula

Cofactor

EC Number Cofactor Comment Organism Structure
1.1.1.35 NAD+
-
 Metallosphaera sedula
4.2.1.17 NADH
-
 Metallosphaera sedula

General Information

EC Number General Information Comment Organism
1.1.1.35 physiological function the enzyme is involved in the 3-hydroxypropionate/4-hydroxybutyrate carbon fixation pathway Metallosphaera sedula
2.3.1.9 physiological function the enzyme is involved in the 3-hydroxypropionate/4-hydroxybutyrate carbon fixation pathway Metallosphaera sedula
4.2.1.120 physiological function the enzyme is involved in the 3-hydroxypropionate/4-hydroxybutyrate carbon fixation pathway Metallosphaera sedula
4.2.1.150 physiological function the enzyme is involved in the 3-hydroxypropionate/4-hydroxybutyrate carbon fixation pathway Metallosphaera sedula
6.2.1.40 physiological function the enzyme is involved in the 3-hydroxypropionate/4-hydroxybutyrate carbon fixation pathway. The flux between the succinate and acetyl-CoA branches in the 3-hydroxypropionate/4-hydroxybutyrate carbon fixation pathway is governed by 4-hydroxybutyrate-CoA ligase, possibly regulated posttranslationally by the protein acetyltransferase Metallosphaera sedula

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
1.1.1.35 260
-
 (S)-3-hydroxybutanoyl-CoA pH 8.0, 70°C Metallosphaera sedula
2.3.1.9 5600
-
 acetoacetyl-CoA pH 7.5, 70°C Metallosphaera sedula
4.2.1.17 260
-
 crotonoyl-CoA at pH 7.0 and 70°C Metallosphaera sedula
4.2.1.17 260
-
 (S)-3-hydroxybutyryl-CoA at pH 7.0 and 70°C Metallosphaera sedula
4.2.1.120 14
-
 4-hydroxybutanoyl-CoA pH 8.0, 70°C Metallosphaera sedula
4.2.1.150 260
-
 crotonyl-CoA pH 8.0, 70°C Metallosphaera sedula
6.2.1.40 0.9
-
 4-hydroxybutanoate pH 7.5, 70°C Metallosphaera sedula