Cloned (Comment) | Organism |
---|---|
recombinant overexpression of His-tagged enzyme in Escherichia coli strain BL21(DE3) | Cupriavidus necator |
Crystallization (Comment) | Organism |
---|---|
purified recombinant enzyme, sitting drop vapour diffusion method, mixing of 0.001 ml of 140 mg/ml protein in 40 mM Tris-HCl, pH 8.0, with 0.001 ml of reservoir solution containing 17% PEG 8000, 0.1 M HEPES pH 7.0, and equilibration against 0.5 ml of reservoir solution, 20°C, 7 days, X-ray diffraction structure determination and analysis at 1.4 A resolution | Cupriavidus necator |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.158 | - |
acetoacetyl-CoA | pH 8.3, 30°C, recombinant enzyme | Cupriavidus necator |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | required | Cupriavidus necator |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
40000 | - |
2 * 40000, recombinant His-tagged enzyme, SDS-PAGE | Cupriavidus necator |
80000 | - |
recombinant His-tagged enzyme, gel filtration | Cupriavidus necator |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
2 acetyl-CoA | Cupriavidus necator | - |
CoA + acetoacetyl-CoA | - |
r | |
2 acetyl-CoA | Cupriavidus necator H16 / ATCC 23440 / NCIB 10442 / S-10-1 | - |
CoA + acetoacetyl-CoA | - |
r | |
CoA + acetoacetyl-CoA | Cupriavidus necator | - |
2 acetyl-CoA | - |
r | |
CoA + acetoacetyl-CoA | Cupriavidus necator H16 / ATCC 23440 / NCIB 10442 / S-10-1 | - |
2 acetyl-CoA | - |
r | |
additional information | Cupriavidus necator | the enzyme shows degradative thiolase activity by converting 3-ketoacyl-CoA to acyl-CoA | ? | - |
? | |
additional information | Cupriavidus necator H16 / ATCC 23440 / NCIB 10442 / S-10-1 | the enzyme shows degradative thiolase activity by converting 3-ketoacyl-CoA to acyl-CoA | ? | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Cupriavidus necator | Q0KAI3 | gene H16_A1887; gene H16_A1887 | - |
Cupriavidus necator H16 / ATCC 23440 / NCIB 10442 / S-10-1 | Q0KAI3 | gene H16_A1887; gene H16_A1887 | - |
Purification (Comment) | Organism |
---|---|
recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) to homogeneity by nickel affinity chromatography and gel filtration | Cupriavidus necator |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
2 acetyl-CoA | - |
Cupriavidus necator | CoA + acetoacetyl-CoA | - |
r | |
2 acetyl-CoA | - |
Cupriavidus necator H16 / ATCC 23440 / NCIB 10442 / S-10-1 | CoA + acetoacetyl-CoA | - |
r | |
CoA + acetoacetyl-CoA | - |
Cupriavidus necator | 2 acetyl-CoA | - |
r | |
CoA + acetoacetyl-CoA | - |
Cupriavidus necator H16 / ATCC 23440 / NCIB 10442 / S-10-1 | 2 acetyl-CoA | - |
r | |
additional information | the enzyme shows degradative thiolase activity by converting 3-ketoacyl-CoA to acyl-CoA | Cupriavidus necator | ? | - |
? | |
additional information | the enzyme shows degradative thiolase activity by converting 3-ketoacyl-CoA to acyl-CoA | Cupriavidus necator H16 / ATCC 23440 / NCIB 10442 / S-10-1 | ? | - |
? |
Subunits | Comment | Organism |
---|---|---|
homodimer | 2 * 40000, recombinant His-tagged enzyme, SDS-PAGE | Cupriavidus necator |
Synonyms | Comment | Organism |
---|---|---|
3-ketoacyl-CoA thiolase | - |
Cupriavidus necator |
A1887 | - |
Cupriavidus necator |
ACAT | - |
Cupriavidus necator |
acetoacetyl-CoA thiolase | - |
Cupriavidus necator |
KACT | - |
Cupriavidus necator |
type II thiolase | - |
Cupriavidus necator |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
30 | - |
assay at | Cupriavidus necator |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
5000000 | - |
acetoacetyl-CoA | pH 8.3, 30°C, recombinant enzyme | Cupriavidus necator |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
8.3 | - |
assay at | Cupriavidus necator |
General Information | Comment | Organism |
---|---|---|
metabolism | the enzyme shows degradative thiolase activity catalyzing the fourth step of beta-oxidation degradative pathways by converting 3-ketoacyl-CoA to acyl-CoA | Cupriavidus necator |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
31650 | - |
acetoacetyl-CoA | pH 8.3, 30°C, recombinant enzyme | Cupriavidus necator |