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Literature summary extracted from

  • Takenoya, M.; Taguchi, S.; Yajima, S.
    Crystal structure and kinetic analyses of a hexameric form of (S)-3-hydroxybutyryl-CoA dehydrogenase from Clostridium acetobutylicum (2018), Acta Crystallogr. Sect. F, 74, 733-740 .
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.1.1.35 expressed in Escherichia coli Rosetta 2(DE3) cells Clostridium acetobutylicum
1.1.1.157 expressed in Escherichia coli Rosetta 2(DE3) cells Clostridium acetobutylicum

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
1.1.1.35 apoenzyme, hanging drop vapor diffusion method, using 1.25% (w/v) PEG 400, 2.2 M ammonium sulfate, 0.1 M HEPES-NaOH pH 7.5. Enzyme in complex with NAD+, hanging drop vapor diffusion method, using 17.5% (w/v) PEG 3350, 200 mM sodium thiocyanate Clostridium acetobutylicum
1.1.1.157 apo and NAD+-bound enzyme forms, hanging drop vapor diffusion method. NAD+-bound crystals grow using a reservoir solution consisting of 17.5% (w/v) PEG 3350, 200 mM sodium thiocyanate. Apo crystals grow using a reservoir solution consisting of 1.25% (w/v) PEG 400, 2.2 M ammonium sulfate, 0.1 M HEPES-NaOH pH 7.5 Clostridium acetobutylicum

Protein Variants

EC Number Protein Variants Comment Organism
1.1.1.35 H138A the mutant shows a slightly lower Km value and a significantly lower kcat value than the wild type enzyme Clostridium acetobutylicum
1.1.1.35 N188A the mutation abolishes the activity of the enzyme Clostridium acetobutylicum
1.1.1.35 S117A the mutation abolishes the activity of the enzyme Clostridium acetobutylicum
1.1.1.157 H138A the mutant shows a slightly lower Km value and a significantly lower kcat value than the wild type enzyme Clostridium acetobutylicum
1.1.1.157 N188A the mutant abolishes the enzyme activity Clostridium acetobutylicum
1.1.1.157 S117A the mutant abolishes the enzyme activity Clostridium acetobutylicum

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.1.1.35 0.0096
-
 3-acetoacetyl-CoA mutant enzyme H138A, at pH 7.5 and 37°C Clostridium acetobutylicum
1.1.1.35 0.0168
-
 NADH mutant enzyme H138A, at pH 7.5 and 37°C Clostridium acetobutylicum
1.1.1.35 0.0227
-
 3-acetoacetyl-CoA wild type enzyme, at pH 7.5 and 37°C Clostridium acetobutylicum
1.1.1.35 0.0314
-
 NADH wild type enzyme, at pH 7.5 and 37°C Clostridium acetobutylicum
1.1.1.157 0.0096
-
 3-acetoacetyl-CoA mutant enzyme H138A, at pH 7.5 and 37°C Clostridium acetobutylicum
1.1.1.157 0.0168
-
 NADH mutant enzyme H138A, at pH 7.5 and 37°C Clostridium acetobutylicum
1.1.1.157 0.0227
-
 3-acetoacetyl-CoA wild type enzyme, at pH 7.5 and 37°C Clostridium acetobutylicum
1.1.1.157 0.0314
-
 NADH wild type enzyme, at pH 7.5 and 37°C Clostridium acetobutylicum

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
1.1.1.35 3-acetoacetyl-CoA + NADH + H+ Clostridium acetobutylicum
-
 (S)-3-hydroxybutanoyl-CoA + NAD+
-
 ?
1.1.1.157 3-acetoacetyl-CoA + NADH + H+ Clostridium acetobutylicum
-
 (S)-3-hydroxybutyryl-CoA + NAD+
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
1.1.1.35 Clostridium acetobutylicum
-
-
-
1.1.1.157 Clostridium acetobutylicum
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.1.1.35 Ni column chromatography and Mono Q column chromatography Clostridium acetobutylicum
1.1.1.157 Ni cOmplete column chromatography and Mono Q column chromatography Clostridium acetobutylicum

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.1.1.35 3-acetoacetyl-CoA + NADH + H+
-
 Clostridium acetobutylicum (S)-3-hydroxybutanoyl-CoA + NAD+
-
 ?
1.1.1.157 3-acetoacetyl-CoA + NADH + H+
-
 Clostridium acetobutylicum (S)-3-hydroxybutyryl-CoA + NAD+
-
 ?

Subunits

EC Number Subunits Comment Organism
1.1.1.35 hexamer x-ray crystallography Clostridium acetobutylicum
1.1.1.157 hexamer
-
 Clostridium acetobutylicum

Synonyms

EC Number Synonyms Comment Organism
1.1.1.35 (S)-3-hydroxybutyryl-CoA dehydrogenase
-
 Clostridium acetobutylicum
1.1.1.35 HBD
-
 Clostridium acetobutylicum
1.1.1.157 (S)-3-hydroxybutyryl-CoA dehydrogenase
-
 Clostridium acetobutylicum
1.1.1.157 HBD
-
 Clostridium acetobutylicum

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.1.1.35 93.8
-
 NADH mutant enzyme H138A, at pH 7.5 and 37°C Clostridium acetobutylicum
1.1.1.35 104.3
-
 3-acetoacetyl-CoA mutant enzyme H138A, at pH 7.5 and 37°C Clostridium acetobutylicum
1.1.1.35 8500
-
 NADH wild type enzyme, at pH 7.5 and 37°C Clostridium acetobutylicum
1.1.1.35 13000
-
 3-acetoacetyl-CoA wild type enzyme, at pH 7.5 and 37°C Clostridium acetobutylicum
1.1.1.157 93.8
-
 NADH mutant enzyme H138A, at pH 7.5 and 37°C Clostridium acetobutylicum
1.1.1.157 104.3
-
 3-acetoacetyl-CoA mutant enzyme H138A, at pH 7.5 and 37°C Clostridium acetobutylicum
1.1.1.157 8500
-
 NADH wild type enzyme, at pH 7.5 and 37°C Clostridium acetobutylicum
1.1.1.157 13000
-
 3-acetoacetyl-CoA wild type enzyme, at pH 7.5 and 37°C Clostridium acetobutylicum

Cofactor

EC Number Cofactor Comment Organism Structure
1.1.1.35 NADH
-
 Clostridium acetobutylicum
1.1.1.157 NADH
-
 Clostridium acetobutylicum

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 0.0056
-
 NADH mutant enzyme H138A, at pH 7.5 and 37°C Clostridium acetobutylicum
1.1.1.35 0.011
-
 3-acetoacetyl-CoA mutant enzyme H138A, at pH 7.5 and 37°C Clostridium acetobutylicum
1.1.1.35 0.27
-
 NADH wild type enzyme, at pH 7.5 and 37°C Clostridium acetobutylicum
1.1.1.35 0.63
-
 3-acetoacetyl-CoA wild type enzyme, at pH 7.5 and 37°C Clostridium acetobutylicum
1.1.1.157 5600
-
 NADH mutant enzyme H138A, at pH 7.5 and 37°C Clostridium acetobutylicum
1.1.1.157 11000
-
 3-acetoacetyl-CoA mutant enzyme H138A, at pH 7.5 and 37°C Clostridium acetobutylicum
1.1.1.157 270000
-
 NADH wild type enzyme, at pH 7.5 and 37°C Clostridium acetobutylicum
1.1.1.157 630000
-
 3-acetoacetyl-CoA wild type enzyme, at pH 7.5 and 37°C Clostridium acetobutylicum