Cloned (Comment) | Organism |
---|---|
recombinant enzyme expression | Streptomyces lividans |
Crystallization (Comment) | Organism |
---|---|
purified recombinant enzyme SlAacS complexed with AMP and acetoacetate, hanging drop vapor diffusion method, mixing of 11 mg/ml protein and 1.5fold molar excess of buffered AMP and acetoacetate with a reservoir solution containing 0.7-1.2M K3citrate and 50 mM BTP-HCl, pH 7.0, 14°C, 1 week, X-ray diffraction structure determination and analysis, molecular replacement using full-length Salmonella enterica acetyl-CoA synthetase, PDB ID1PG4, stripped of waters and cofactors/substrates as search model | Streptomyces lividans |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | required | Streptomyces lividans |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + acetoacetate + CoA | Streptomyces lividans | - |
AMP + diphosphate + acetoacetyl-CoA | - |
? | |
ATP + acetoacetate + CoA | Streptomyces lividans TK24 | - |
AMP + diphosphate + acetoacetyl-CoA | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Streptomyces lividans | D6EQU8 | - |
- |
Streptomyces lividans TK24 | D6EQU8 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant enzyme | Streptomyces lividans |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
4.91 | 6.8 | purified recombinant enzyme, pH 7.5, 30°C | Streptomyces lividans |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + acetoacetate + CoA | - |
Streptomyces lividans | AMP + diphosphate + acetoacetyl-CoA | - |
? | |
ATP + acetoacetate + CoA | - |
Streptomyces lividans TK24 | AMP + diphosphate + acetoacetyl-CoA | - |
? |
Synonyms | Comment | Organism |
---|---|---|
AACS | - |
Streptomyces lividans |
Acetoacetyl-CoA synthetase | - |
Streptomyces lividans |
acsA1 | - |
Streptomyces lividans |
SlAacS | - |
Streptomyces lividans |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
30 | - |
assay at | Streptomyces lividans |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
5.89 | - |
acetoacetate | pH 7.5, 30°C, recombinant enzyme | Streptomyces lividans | |
6.43 | - |
ATP | pH 7.5, 30°C, recombinant enzyme | Streptomyces lividans | |
8.16 | - |
CoA | pH 7.5, 30°C, recombinant enzyme | Streptomyces lividans |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.5 | - |
assay at | Streptomyces lividans |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
ATP | - |
Streptomyces lividans |
General Information | Comment | Organism |
---|---|---|
additional information | in addition to the two catalytic states, additional conformations are observed crystallographically that likely play a role in allowing access and egress of substrates and products from the relatively buried active site. The enzyme shows conformational flexibility. Structure-function analysis, overview. The C-terminal domain undergoes a large conformational change in the catalytic mechanism of acyl-CoA synthetases, the C-terminal extension is important for catalytic activity, structure comparisons. One region from the N-terminal domain interacts is the so-called P-loop, a glycine-, serine-, and threonine-rich region that interacts with the phosphates of ATP. This P-loop adopts multiple conformations in the different crystal structures and may play an important role in the release of PPi and trigger the conformational change. Specifically, the main chain carbonyls of Ser272, Gly274, and Gly277 form direct or water-mediated hydrogen bonds with Asn637 and Ser640. Asn637 also interacts directly with Arg183 and Asp187, while the carbonyl of Gly639 and the carbonyl and side chain oxygens of Ser640 interact with Ser184, Asp187, and Arg188. | Streptomyces lividans |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
23.6 | - |
CoA | pH 7.5, 30°C, recombinant enzyme | Streptomyces lividans | |
26.6 | - |
acetoacetate | pH 7.5, 30°C, recombinant enzyme | Streptomyces lividans | |
32.4 | - |
ATP | pH 7.5, 30°C, recombinant enzyme | Streptomyces lividans |