EC Number | Cloned (Comment) | Organism |
---|---|---|
2.3.1.9 | gene ReH16_B0759, DNA and amino acid sequence determination and analysis, recombinant overexpression of His-tagged enzyme in Escherichia coli strain BL21(DE3) | Cupriavidus necator |
EC Number | Crystallization (Comment) | Organism |
---|---|---|
2.3.1.9 | purified recombinant enzyme in apoform and with bound CoA, hanging drop vapor diffusion method, mixing 0.001 ml of 40 mg/ml protein in 40 mM Tris-HCl, pH 8.0, and 5 mM 2-mercaptoethanol, with 0.001 ml of reservoir solution containing 1.0 M ammonium sulfate, 0.1 M HEPES, pH 7.25, and equilibration against 0.5 ml of reservoir solution, 20°C, 3 days, X-ray diffraction structure determination and analysis at 2.0-2.3 A resolution, molecular replacement method using the structure of Mycobacterium tuberculosis thiolase Mttt0182, PDB ID1ULQ, as a search model, structure modeling | Cupriavidus necator |
EC Number | Protein Variants | Comment | Organism |
---|---|---|---|
2.3.1.9 | C377A | site-directed mutagenesis, almost inactive mutant | Cupriavidus necator |
2.3.1.9 | C89A | site-directed mutagenesis, almost inactive mutant | Cupriavidus necator |
2.3.1.9 | F156A | site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme | Cupriavidus necator |
2.3.1.9 | H347A | site-directed mutagenesis, almost inactive mutant | Cupriavidus necator |
2.3.1.9 | K17A | site-directed mutagenesis, the mutant shows 60% reduced activity compared to the wild-type enzyme | Cupriavidus necator |
2.3.1.9 | M124A | site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme | Cupriavidus necator |
2.3.1.9 | R220A | site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme | Cupriavidus necator |
2.3.1.9 | V231A | site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme | Cupriavidus necator |
EC Number | KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
2.3.1.9 | additional information | - |
additional information | Michaelis-Menten kinetics | Cupriavidus necator |
EC Number | Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|---|
2.3.1.9 | CoA + acetoacetyl-CoA | Cupriavidus necator | - |
2 acetyl-CoA | - |
? | |
2.3.1.9 | CoA + acetoacetyl-CoA | Cupriavidus necator H16 / ATCC 23440 / NCIB 10442 / S-10-1 | - |
2 acetyl-CoA | - |
? | |
2.3.1.9 | additional information | Cupriavidus necator | the enzyme shows degradative thiolase activity by converting 3-ketoacyl-CoA to acyl-CoA | ? | - |
? | |
2.3.1.9 | 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 | ? | - |
? |
EC Number | Organism | UniProt | Comment | Textmining |
---|---|---|---|---|
2.3.1.9 | Cupriavidus necator | Q0K368 | gene H16_B0759; gene H16_B0759 | - |
2.3.1.9 | Cupriavidus necator H16 / ATCC 23440 / NCIB 10442 / S-10-1 | Q0K368 | gene H16_B0759; gene H16_B0759 | - |
EC Number | Purification (Comment) | Organism |
---|---|---|
2.3.1.9 | recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) to about 95% purity by nickel affinity chromatography and gel filtration | Cupriavidus necator |
EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|---|
2.3.1.9 | CoA + acetoacetyl-CoA | - |
Cupriavidus necator | 2 acetyl-CoA | - |
? | |
2.3.1.9 | CoA + acetoacetyl-CoA | CoA substrate binding structure, overview | Cupriavidus necator | 2 acetyl-CoA | - |
? | |
2.3.1.9 | CoA + acetoacetyl-CoA | - |
Cupriavidus necator H16 / ATCC 23440 / NCIB 10442 / S-10-1 | 2 acetyl-CoA | - |
? | |
2.3.1.9 | CoA + acetoacetyl-CoA | CoA substrate binding structure, overview | Cupriavidus necator H16 / ATCC 23440 / NCIB 10442 / S-10-1 | 2 acetyl-CoA | - |
? | |
2.3.1.9 | additional information | the enzyme shows degradative thiolase activity by converting 3-ketoacyl-CoA to acyl-CoA | Cupriavidus necator | ? | - |
? | |
2.3.1.9 | additional information | the bound CoA substrate is located within the deep cleft formed by subdomains I, II, and III. The thiol group of CoA is located in the vicinity of the catalytic site for the degradation reaction, whereas,the adenosine nucleotide moiety is exposed on the surface. Unlike enzyme ReH16_A1887, that shows substantial structural changeupon the binding of CoA, the CoA-bound form of enzyme ReH16_B0759 exhibits quite similar structure to the apoform of the protein except for residual movement of Arg220. Moreover, the CoA binding mode of enzyme ReH16_B0759 is quite different from that of enzyme ReH16_A1887 | Cupriavidus necator | ? | - |
? | |
2.3.1.9 | 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 | ? | - |
? | |
2.3.1.9 | additional information | the bound CoA substrate is located within the deep cleft formed by subdomains I, II, and III. The thiol group of CoA is located in the vicinity of the catalytic site for the degradation reaction, whereas,the adenosine nucleotide moiety is exposed on the surface. Unlike enzyme ReH16_A1887, that shows substantial structural changeupon the binding of CoA, the CoA-bound form of enzyme ReH16_B0759 exhibits quite similar structure to the apoform of the protein except for residual movement of Arg220. Moreover, the CoA binding mode of enzyme ReH16_B0759 is quite different from that of enzyme ReH16_A1887 | Cupriavidus necator H16 / ATCC 23440 / NCIB 10442 / S-10-1 | ? | - |
? |
EC Number | Synonyms | Comment | Organism |
---|---|---|---|
2.3.1.9 | 3-ketoacyl-CoA thiolase | - |
Cupriavidus necator |
2.3.1.9 | 3-ketoacyl-coenzyme A thiolase | - |
Cupriavidus necator |
2.3.1.9 | ACAT | - |
Cupriavidus necator |
2.3.1.9 | KACT | - |
Cupriavidus necator |
2.3.1.9 | ReH16_B0759 | - |
Cupriavidus necator |
2.3.1.9 | type II thiolase | - |
Cupriavidus necator |
EC Number | General Information | Comment | Organism |
---|---|---|---|
2.3.1.9 | 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 |
2.3.1.9 | additional information | similar to other degradative thiolases, enzyme ReH16_B0759 functions as a dimer, and the monomer comprises three subdomains. Unlike enzyme ReH16_A1887, a substantial structural change is not observed upon the binding of the CoA substrate in enzyme ReH16_B0759. At the active site of the enzyme highly conserved residues Cys89, His347, and Cys377are located near the thiol-group of CoA | Cupriavidus necator |