Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli BL21(DE3) cells and in Corynebacterium glutamicum | Cupriavidus necator |
Crystallization (Comment) | Organism |
---|---|
vapor diffusion method, using 0.1 M MES (pH 7.1), 1.6 M ammonium sulfate, and 10% (w/v) 1,4-dioxane | Cupriavidus necator |
Protein Variants | Comment | Organism |
---|---|---|
Q47L | the mutant with increased specific activity exhibits a kcat value that is 2.4fold higher than that of the wild type enzyme | Cupriavidus necator |
T173S | the mutant with increased specific activity exhibits a kcat value that is 3.5fold higher than that of the wild type enzyme | Cupriavidus necator |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0057 | - |
acetoacetyl-CoA | wild type enzyme, at pH 8.0 and 30°C | Cupriavidus necator | |
0.0136 | - |
acetoacetyl-CoA | mutant enzyme Q47L, at pH 8.0 and 30°C | Cupriavidus necator | |
0.0159 | - |
acetoacetyl-CoA | mutant enzyme Q47L, at pH 8.0 and 30°C | Cupriavidus necator | |
0.149 | - |
NADPH | wild type enzyme, at pH 8.0 and 30°C | Cupriavidus necator | |
0.289 | - |
NADPH | mutant enzyme Q47L, at pH 8.0 and 30°C | Cupriavidus necator | |
0.617 | - |
NADPH | mutant enzyme Q47L, at pH 8.0 and 30°C | Cupriavidus necator |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
acetoacetyl-CoA + NADPH + H+ | Cupriavidus necator | - |
3-hydroxybutyryl-CoA + NADP+ | - |
? | |
acetoacetyl-CoA + NADPH + H+ | Cupriavidus necator H16 / ATCC 23440 / NCIB 10442 / S-10-1 | - |
3-hydroxybutyryl-CoA + NADP+ | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Cupriavidus necator | P14697 | - |
- |
Cupriavidus necator H16 / ATCC 23440 / NCIB 10442 / S-10-1 | P14697 | - |
- |
Purification (Comment) | Organism |
---|---|
His-Bind resin column chromatography and Superdex 200 pg gel filtration | Cupriavidus necator |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
acetoacetyl-CoA + NADPH + H+ | - |
Cupriavidus necator | 3-hydroxybutyryl-CoA + NADP+ | - |
? | |
acetoacetyl-CoA + NADPH + H+ | - |
Cupriavidus necator H16 / ATCC 23440 / NCIB 10442 / S-10-1 | 3-hydroxybutyryl-CoA + NADP+ | - |
? |
Synonyms | Comment | Organism |
---|---|---|
NADPH-dependent acetoacetyl coenzyme A reductase | - |
Cupriavidus necator |
PhaB | - |
Cupriavidus necator |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
102 | - |
NADPH | wild type enzyme, at pH 8.0 and 30°C | Cupriavidus necator | |
102 | - |
acetoacetyl-CoA | wild type enzyme, at pH 8.0 and 30°C | Cupriavidus necator | |
249 | - |
NADPH | mutant enzyme Q47L, at pH 8.0 and 30°C | Cupriavidus necator | |
249 | - |
acetoacetyl-CoA | mutant enzyme Q47L, at pH 8.0 and 30°C | Cupriavidus necator | |
361 | - |
NADPH | mutant enzyme Q47L, at pH 8.0 and 30°C | Cupriavidus necator | |
361 | - |
acetoacetyl-CoA | mutant enzyme Q47L, at pH 8.0 and 30°C | Cupriavidus necator |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NADP+ | - |
Cupriavidus necator |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
585 | - |
NADPH | mutant enzyme Q47L, at pH 8.0 and 30°C | Cupriavidus necator | |
685 | - |
NADPH | wild type enzyme, at pH 8.0 and 30°C | Cupriavidus necator | |
862 | - |
NADPH | mutant enzyme Q47L, at pH 8.0 and 30°C | Cupriavidus necator | |
15200 | - |
acetoacetyl-CoA | mutant enzyme Q47L, at pH 8.0 and 30°C | Cupriavidus necator | |
18000 | - |
acetoacetyl-CoA | wild type enzyme, at pH 8.0 and 30°C | Cupriavidus necator | |
26500 | - |
acetoacetyl-CoA | mutant enzyme Q47L, at pH 8.0 and 30°C | Cupriavidus necator |