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Literature summary for 2.7.2.1 extracted from
- Structural and mutational analysis of amino acid residues involved in ATP specificity of Escherichia coli acetate kinase (2014), J. Biosci. Bioeng., 118, 502-507.
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| gene ackA, sequence comparisons, recombinant expression of wild-type and mutant enzymes in Escherichia coli strain DH5alpha | Escherichia coli |
Protein Variants
| Protein Variants | Comment | Organism |
|---|---|---|
| G332D | site-directed mutagenesis, the mutant shows increased Km for ATP and reduced ATP-dependent acetate kinase activity compared to the wild-type enzyme | Escherichia coli |
| G333Q | site-directed mutagenesis, the mutant shows increased Km for ATP and reduced ATP-dependent acetate kinase activity compared to the wild-type enzyme | Escherichia coli |
| I334M | site-directed mutagenesis, the mutant shows increased Km for ATP and reduced ATP-dependent acetate kinase activity compared to the wild-type enzyme | Escherichia coli |
| additional information | each of five candidate residue in Escherichia coli ATP-specific AK (ATP-ecoAK), which is unable to use diphosphate, is substituted with the respective diphosphate-ehiAK amino acid residue. Each variant ATP-ecoAK has an increased Km for ATP, indicating that the five residues are the determinants for the specificity to ATP in ATP-ecoAK | Escherichia coli |
| N213T | site-directed mutagenesis, the mutant shows increased Km for ATP and reduced ATP-dependent acetate kinase activity compared to the wild-type enzyme | Escherichia coli |
| N213T/G332D/E336L/T385N | site-directed mutagenesis, the mutant shows unaltered Km for ATP and highly reduced ATP-dependent acetate kinase activity compared to the wild-type enzyme | Escherichia coli |
| N337E | site-directed mutagenesis, the mutant shows increased Km for ATP and reduced ATP-dependent acetate kinase activity compared to the wild-type enzyme | Escherichia coli |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| additional information | - |
additional information | Michaelis-Menten kinetics | Escherichia coli | |
| 0.66 | - |
ATP | pH 6.5, 30°C, recombinant mutant N213T/G332D/E336L/T385N | Escherichia coli |  |
| 0.67 | - |
ATP | pH 6.5, 30°C, recombinant wild-type enzyme | Escherichia coli | |
| 1.7 | - |
ATP | pH 6.5, 30°C, recombinant mutant N213T | Escherichia coli | |
| 3.5 | - |
ATP | pH 6.5, 30°C, recombinant mutant G332D | Escherichia coli | |
| 5.2 | - |
ATP | pH 6.5, 30°C, recombinant mutant G333Q | Escherichia coli | |
| 6.6 | - |
ATP | pH 6.5, 30°C, recombinant mutant I334M | Escherichia coli | |
| 30.7 | - |
ATP | pH 6.5, 30°C, recombinant mutant N337E | Escherichia coli | |
Metals/Ions
| Metals/Ions | Comment | Organism | Structure |
|---|---|---|---|
| Mg2+ | required | Escherichia coli | |
Natural Substrates/ Products (Substrates)
| Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| ATP + acetate | Escherichia coli | - |
ADP + acetyl phosphate | - |
r | |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Escherichia coli | P0A6A3 | gene ackA | - |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| recombinant wild-type and mutant enzymes from Escherichia coli strain DH5alpha by ATP affinity chromatography | Escherichia coli |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| acetyl phosphate + hydroxylamine | - |
Escherichia coli | acetyl hydroxamate + phosphate | - |
? | |
| ATP + acetate | - |
Escherichia coli | ADP + acetyl phosphate | - |
r | |
| additional information | no activity with diphosphate | Escherichia coli | ? | - |
? | |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| ackA | - |
Escherichia coli |
| ATP-ecoAK | - |
Escherichia coli |
| ATP-specific AK | - |
Escherichia coli |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 30 | - |
assay at | Escherichia coli |
Turnover Number [1/s]
| Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.34 | - |
ATP | pH 6.5, 30°C, recombinant mutant N213T/G332D/E336L/T385N | Escherichia coli | |
| 9.9 | - |
ATP | pH 6.5, 30°C, recombinant mutant G332D | Escherichia coli | |
| 10.5 | - |
ATP | pH 6.5, 30°C, recombinant mutant I334M | Escherichia coli | |
| 16.3 | - |
ATP | pH 6.5, 30°C, recombinant mutant N213T | Escherichia coli | |
| 41 | - |
ATP | pH 6.5, 30°C, recombinant mutant G333Q | Escherichia coli | |
| 395 | - |
ATP | pH 6.5, 30°C, recombinant mutant N337E | Escherichia coli | |
| 908 | - |
ATP | pH 6.5, 30°C, recombinant wild-type enzyme | Escherichia coli | |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 6.5 | 7.5 | acetyl phosphate-forming reaction direction, assay at | Escherichia coli |
Cofactor
| Cofactor | Comment | Organism | Structure |
|---|---|---|---|
| ADP | - |
Escherichia coli | |
| ATP | - |
Escherichia coli | |
| additional information | no activity with diphosphate as phosphoryl donor | Escherichia coli |
General Information
| General Information | Comment | Organism |
|---|---|---|
| evolution | residue Asn337 of ATP-ecoAK is particularly significant for the specificity to ATP. The 5 residues are highly conserved in 2625 PPi-ehiAK homologsue implying that almost all organisms have ATP-dependent acetate kinase, EC 2.7.2.1, rather than diphosphate-dependent acetate kinase, EC 2.7.2.12 | Escherichia coli |
| additional information | substrate-binding site structure and comparison with diphosphate-dependent acetate kinase, EC 2.7.2.12, overview | Escherichia coli |
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