Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli Rosetta 2(DE3) cells | Clostridium acetobutylicum |
Crystallization (Comment) | Organism |
---|---|
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 |
Protein Variants | Comment | Organism |
---|---|---|
H138A | the mutant shows a slightly lower Km value and a significantly lower kcat value than the wild type enzyme | Clostridium acetobutylicum |
N188A | the mutation abolishes the activity of the enzyme | Clostridium acetobutylicum |
S117A | the mutation abolishes the activity of the enzyme | Clostridium acetobutylicum |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0096 | - |
3-acetoacetyl-CoA | mutant enzyme H138A, at pH 7.5 and 37°C | Clostridium acetobutylicum | |
0.0168 | - |
NADH | mutant enzyme H138A, at pH 7.5 and 37°C | Clostridium acetobutylicum | |
0.0227 | - |
3-acetoacetyl-CoA | wild type enzyme, at pH 7.5 and 37°C | Clostridium acetobutylicum | |
0.0314 | - |
NADH | wild type enzyme, at pH 7.5 and 37°C | Clostridium acetobutylicum |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
3-acetoacetyl-CoA + NADH + H+ | Clostridium acetobutylicum | - |
(S)-3-hydroxybutanoyl-CoA + NAD+ | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Clostridium acetobutylicum | - |
- |
- |
Purification (Comment) | Organism |
---|---|
Ni column chromatography and Mono Q column chromatography | Clostridium acetobutylicum |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
3-acetoacetyl-CoA + NADH + H+ | - |
Clostridium acetobutylicum | (S)-3-hydroxybutanoyl-CoA + NAD+ | - |
? |
Subunits | Comment | Organism |
---|---|---|
hexamer | x-ray crystallography | Clostridium acetobutylicum |
Synonyms | Comment | Organism |
---|---|---|
(S)-3-hydroxybutyryl-CoA dehydrogenase | - |
Clostridium acetobutylicum |
HBD | - |
Clostridium acetobutylicum |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
93.8 | - |
NADH | mutant enzyme H138A, at pH 7.5 and 37°C | Clostridium acetobutylicum | |
104.3 | - |
3-acetoacetyl-CoA | mutant enzyme H138A, at pH 7.5 and 37°C | Clostridium acetobutylicum | |
8500 | - |
NADH | wild type enzyme, at pH 7.5 and 37°C | Clostridium acetobutylicum | |
13000 | - |
3-acetoacetyl-CoA | wild type enzyme, at pH 7.5 and 37°C | Clostridium acetobutylicum |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NADH | - |
Clostridium acetobutylicum |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0056 | - |
NADH | mutant enzyme H138A, at pH 7.5 and 37°C | Clostridium acetobutylicum | |
0.011 | - |
3-acetoacetyl-CoA | mutant enzyme H138A, at pH 7.5 and 37°C | Clostridium acetobutylicum | |
0.27 | - |
NADH | wild type enzyme, at pH 7.5 and 37°C | Clostridium acetobutylicum | |
0.63 | - |
3-acetoacetyl-CoA | wild type enzyme, at pH 7.5 and 37°C | Clostridium acetobutylicum |