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Literature summary for 3.5.1.135 extracted from
- YqfB protein from Escherichia coli an atypical amidohydrolase active towards N4-acylcytosine derivatives (2020), Sci. Rep., 10, 788 .
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| expressed in Escherichia coli BL21(DE3) cells | Escherichia coli |
Protein Variants
| Protein Variants | Comment | Organism |
|---|---|---|
| D27A | the mutant shows severely reduced activity compared to the wild type enzyme | Escherichia coli |
| E74A | with N4-acetylcytidine and N4-acetylcytosine, the turnover number of the mutant is 1000fold and 100fold lower, respectively, than that of the wild type enzyme | Escherichia coli |
| H70A | more than 10fold decrease in activity towards N4-acetylcytidine and N4-acetylcytosine is observed for the mutant enzyme | Escherichia coli |
| K21A | catalytically inactive | Escherichia coli |
| R26A | catalytically inactive | Escherichia coli |
| T24A | catalytically inactive | Escherichia coli |
| Y89A | more than 3fold decrease in activity towards N4-acetylcytidine and N4-acetylcytosine is observed for the mutant enzyme | Escherichia coli |
Inhibitors
| Inhibitors | Comment | Organism | Structure |
|---|---|---|---|
| additional information | the catalytic activity of the enzyme is unaffected by phenylmethylsulfonyl fluoride, p-hydroxymercuribenzoate (up to 2 mM), p-chloromercuribenzoate (up to 1 mM), and EDTA (up to 5 mM) | Escherichia coli | |
| N4-acetyl-5-fluorocytosine | - |
Escherichia coli |  |
| N4-acetylcytidine | - |
Escherichia coli | |
| N4-acetylcytosine | - |
Escherichia coli | |
| N4-benzoyl-2'-deoxycytidine | - |
Escherichia coli | |
| N4-benzoylcytidine | - |
Escherichia coli | |
| N4-hexanoyl-2'-deoxycytidine | - |
Escherichia coli | |
| N4-isobutyryl-2'-deoxycytidine | - |
Escherichia coli | |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.002 | - |
N4-acetylcytidine | mutant enzyme E74A, at pH 8.0 and 22°C | Escherichia coli | |
| 0.05 | - |
N4-acetylcytidine | mutant enzyme H70A, at pH 8.0 and 22°C | Escherichia coli | |
| 0.062 | - |
N4-acetylcytidine | recombinant C-terminally His6-tagged enzyme, at pH 8.0 and 22°C | Escherichia coli | |
| 0.07 | - |
N4-acetylcytosine | recombinant C-terminally His6-tagged enzyme, at pH 8.0 and 22°C | Escherichia coli | |
| 0.07 | - |
N4-acetylcytidine | recombinant enzyme, at pH 8.0 and 22°C | Escherichia coli | |
| 0.07 | - |
N4-acetylcytidine | recombinant N-terminally His6-tagged enzyme, at pH 8.0 and 22°C | Escherichia coli | |
| 0.14 | - |
N4-hexanoyl-2'-deoxycytidine | recombinant C-terminally His6-tagged enzyme, at pH 8.0 and 22°C | Escherichia coli | |
| 0.29 | - |
N4-acetylcytidine | mutant enzyme D27A, at pH 8.0 and 22°C | Escherichia coli | |
| 0.3 | - |
N4-acetylcytosine | mutant enzyme E74A, at pH 8.0 and 22°C | Escherichia coli | |
| 0.3 | - |
N4-benzoyl-2'-deoxycytidine | recombinant C-terminally His6-tagged enzyme, at pH 8.0 and 22°C | Escherichia coli | |
| 0.4 | - |
N4-acetyl-2'-deoxycytidine | recombinant C-terminally His6-tagged enzyme, at pH 8.0 and 22°C | Escherichia coli | |
| 0.7 | - |
capecitabine | recombinant C-terminally His6-tagged enzyme, at pH 8.0 and 22°C | Escherichia coli | |
| 1.9 | - |
N4-isobutyryl-2'-deoxycytidine | recombinant C-terminally His6-tagged enzyme, at pH 8.0 and 22°C | Escherichia coli | |
| 2.1 | - |
N4-acetyl-5-fluorocytosine | recombinant C-terminally His6-tagged enzyme, at pH 8.0 and 22°C | Escherichia coli | |
| 2.2 | - |
N4-acetylcytidine | mutant enzyme Y89A, at pH 8.0 and 22°C | Escherichia coli | |
| 5.1 | - |
N4-benzoylcytidine | recombinant C-terminally His6-tagged enzyme, at pH 8.0 and 22°C | Escherichia coli | |
Natural Substrates/ Products (Substrates)
| Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| N4-acetylcytidine + H2O | Escherichia coli | - |
cytidine + acetate | - |
? | |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Escherichia coli | P67603 | - |
- |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| Q Sepharose column chromatography, ANX Sepharose column chromatography, phenyl Sepharose column chromatography, Resource ISO, and Source 15Q column chromatography | Escherichia coli |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| capecitabine + H2O | - |
Escherichia coli | ? | - |
? | |
| additional information | the enzyme does not hydrolyze N4-acetyl-2',3',5'-tri-O-acetylcytidine, N4-(2-acetyl-benzoyl)-2'-deoxycytidine, N4-(3-benzoyl-benzoyl)-2'-deoxycytidine, N4-(2-acetyl-benzoyl)-2'-deoxycytidine-5'-triphosphate, N4-(3-benzoyl-benzoyl)-2'-deoxycytidine-5'-triphosphate, N2-acetylisocytosine, 2-acetylaminopyridine, 4-nitroacetanilide and 4-nitrophenyl acetate | Escherichia coli | ? | - |
- |
|
| N4-(3-acetyl-benzoyl)-2'-deoxycytidine + H2O | - |
Escherichia coli | ? | - |
? | |
| N4-(3-acetyl-benzoyl)-2'-deoxycytidine-5'-triphosphate + H2O | - |
Escherichia coli | ? | - |
? | |
| N4-(4-acetyl-benzoyl)-2'-deoxycytidine + H2O | - |
Escherichia coli | ? | - |
? | |
| N4-(4-acetyl-benzoyl)-2'-deoxycytidine-5'-triphosphate + H2O | - |
Escherichia coli | ? | - |
? | |
| N4-(4-benzoyl-benzoyl)-2'-deoxycytidine + H2O | - |
Escherichia coli | ? | - |
? | |
| N4-(4-benzoyl-benzoyl)-2'-deoxycytidine-5'-triphosphate + H2O | - |
Escherichia coli | ? | - |
? | |
| N4-acetyl-2'-deoxy-5'-O-DMT-cytidine + H2O | - |
Escherichia coli | ? | - |
? | |
| N4-acetyl-2'-deoxycytidine + H2O | - |
Escherichia coli | ? | - |
? | |
| N4-acetyl-2'-deoxycytidine-5'-triphosphate + H2O | - |
Escherichia coli | ? | - |
? | |
| N4-acetyl-5-fluorocytosine + H2O | - |
Escherichia coli | ? | - |
? | |
| N4-acetylcytidine + H2O | - |
Escherichia coli | cytidine + acetate | - |
? | |
| N4-acetylcytosine + H2O | - |
Escherichia coli | ? | - |
? | |
| N4-benzoyl-2'-deoxycytidine + H2O | - |
Escherichia coli | ? | - |
? | |
| N4-benzoyl-2'-deoxycytidine-5'-triphosphate + H2O | - |
Escherichia coli | ? | - |
? | |
| N4-benzoylcytidine + H2O | - |
Escherichia coli | ? | - |
? | |
| N4-hexanoyl-2'-deoxycytidine + H2O | - |
Escherichia coli | ? | - |
? | |
| N4-hexanoyl-2'-deoxycytidine-5'-triphosphate + H2O | - |
Escherichia coli | ? | - |
? | |
| N4-isobutyryl-2'-deoxycytidine + H2O | - |
Escherichia coli | ? | - |
? | |
| N4-nicotinoyl-2'-deoxycytidine + H2O | - |
Escherichia coli | ? | - |
? | |
| N4-nicotinoyl-2'-deoxycytidine-5'-triphosphate + H2O | - |
Escherichia coli | ? | - |
? | |
Subunits
| Subunits | Comment | Organism |
|---|---|---|
| monomer | 1 * 14000, SDS-PAGE | Escherichia coli |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| yqfB | - |
Escherichia coli |
Turnover Number [1/s]
| Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.016 | - |
capecitabine | recombinant C-terminally His6-tagged enzyme, at pH 8.0 and 22°C | Escherichia coli | |
| 0.07 | - |
N4-acetylcytidine | mutant enzyme E74A, at pH 8.0 and 22°C | Escherichia coli | |
| 0.07 | - |
N4-benzoyl-2'-deoxycytidine | recombinant C-terminally His6-tagged enzyme, at pH 8.0 and 22°C | Escherichia coli | |
| 0.22 | - |
N4-acetylcytosine | mutant enzyme E74A, at pH 8.0 and 22°C | Escherichia coli | |
| 1.8 | - |
N4-benzoylcytidine | recombinant C-terminally His6-tagged enzyme, at pH 8.0 and 22°C | Escherichia coli | |
| 23 | - |
N4-acetylcytidine | mutant enzyme H70A, at pH 8.0 and 22°C | Escherichia coli | |
| 24 | - |
N4-hexanoyl-2'-deoxycytidine | recombinant C-terminally His6-tagged enzyme, at pH 8.0 and 22°C | Escherichia coli | |
| 40 | - |
N4-acetylcytidine | mutant enzyme Y89A, at pH 8.0 and 22°C | Escherichia coli | |
| 70 | - |
N4-acetylcytosine | recombinant C-terminally His6-tagged enzyme, at pH 8.0 and 22°C | Escherichia coli | |
| 75 | - |
N4-isobutyryl-2'-deoxycytidine | recombinant C-terminally His6-tagged enzyme, at pH 8.0 and 22°C | Escherichia coli | |
| 90 | - |
N4-acetyl-5-fluorocytosine | recombinant C-terminally His6-tagged enzyme, at pH 8.0 and 22°C | Escherichia coli | |
| 101 | - |
N4-acetyl-2'-deoxycytidine | recombinant C-terminally His6-tagged enzyme, at pH 8.0 and 22°C | Escherichia coli | |
| 154 | - |
N4-acetylcytidine | recombinant N-terminally His6-tagged enzyme, at pH 8.0 and 22°C | Escherichia coli | |
| 157 | - |
N4-acetylcytidine | recombinant C-terminally His6-tagged enzyme, at pH 8.0 and 22°C | Escherichia coli | |
| 196 | - |
N4-acetylcytidine | recombinant enzyme, at pH 8.0 and 22°C | Escherichia coli | |
| 300 | - |
N4-acetylcytidine | mutant enzyme D27A, at pH 8.0 and 22°C | Escherichia coli | |
Ki Value [mM]
| Ki Value [mM] | Ki Value maximum [mM] | Inhibitor | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.0003 | - |
N4-acetylcytidine | mutant enzyme E74A, at pH 8.0 and 22°C | Escherichia coli | |
| 0.00085 | - |
N4-benzoyl-2'-deoxycytidine | recombinant C-terminally His6-tagged enzyme, at pH 8.0 and 22°C | Escherichia coli | |
| 0.077 | - |
N4-acetylcytidine | recombinant enzyme, at pH 8.0 and 22°C | Escherichia coli | |
| 0.13 | - |
N4-acetylcytidine | recombinant C-terminally His6-tagged enzyme, at pH 8.0 and 22°C | Escherichia coli | |
| 0.13 | - |
N4-acetylcytidine | recombinant N-terminally His6-tagged enzyme, at pH 8.0 and 22°C | Escherichia coli | |
| 0.15 | - |
N4-benzoylcytidine | recombinant C-terminally His6-tagged enzyme, at pH 8.0 and 22°C | Escherichia coli | |
| 0.2 | - |
N4-acetylcytidine | mutant enzyme H70A, at pH 8.0 and 22°C | Escherichia coli | |
| 0.4 | - |
N4-acetylcytidine | mutant enzyme D27A, at pH 8.0 and 22°C | Escherichia coli | |
| 0.7 | - |
N4-acetylcytosine | mutant enzyme Y89A, at pH 8.0 and 22°C | Escherichia coli | |
| 0.7 | - |
N4-acetylcytosine | recombinant C-terminally His6-tagged enzyme, at pH 8.0 and 22°C | Escherichia coli | |
| 1.6 | - |
N4-isobutyryl-2'-deoxycytidine | recombinant C-terminally His6-tagged enzyme, at pH 8.0 and 22°C | Escherichia coli | |
| 2.8 | - |
N4-hexanoyl-2'-deoxycytidine | recombinant C-terminally His6-tagged enzyme, at pH 8.0 and 22°C | Escherichia coli | |
| 4 | - |
N4-acetyl-5-fluorocytosine | recombinant C-terminally His6-tagged enzyme, at pH 8.0 and 22°C | Escherichia coli | |
kcat/KM [mM/s]
| kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.015 | - |
N4-hexanoyl-2'-deoxycytidine-5'-triphosphate | recombinant C-terminally His6-tagged enzyme, at pH 8.0 and 22°C | Escherichia coli | |
| 0.022 | - |
capecitabine | recombinant C-terminally His6-tagged enzyme, at pH 8.0 and 22°C | Escherichia coli | |
| 0.23 | - |
N4-benzoyl-2'-deoxycytidine | recombinant C-terminally His6-tagged enzyme, at pH 8.0 and 22°C | Escherichia coli | |
| 0.32 | - |
N4-acetylcytosine | mutant enzyme Y89A, at pH 8.0 and 22°C | Escherichia coli | |
| 0.7 | - |
N4-acetylcytosine | mutant enzyme E74A, at pH 8.0 and 22°C | Escherichia coli | |
| 3.5 | - |
N4-benzoylcytidine | recombinant C-terminally His6-tagged enzyme, at pH 8.0 and 22°C | Escherichia coli | |
| 7 | - |
N4-acetylcytosine | mutant enzyme H70A, at pH 8.0 and 22°C | Escherichia coli | |
| 19 | - |
N4-acetylcytidine | mutant enzyme Y89A, at pH 8.0 and 22°C | Escherichia coli | |
| 30 | - |
N4-acetylcytidine | mutant enzyme E74A, at pH 8.0 and 22°C | Escherichia coli | |
| 31 | - |
N4-acetylcytosine | mutant enzyme D27A, at pH 8.0 and 22°C | Escherichia coli | |
| 37.5 | - |
N4-isobutyryl-2'-deoxycytidine | recombinant C-terminally His6-tagged enzyme, at pH 8.0 and 22°C | Escherichia coli | |
| 40 | - |
N4-acetyl-5-fluorocytosine | recombinant C-terminally His6-tagged enzyme, at pH 8.0 and 22°C | Escherichia coli | |
| 72 | - |
N4-hexanoyl-2'-deoxycytidine | recombinant C-terminally His6-tagged enzyme, at pH 8.0 and 22°C | Escherichia coli | |
| 100 | - |
N4-acetylcytosine | recombinant C-terminally His6-tagged enzyme, at pH 8.0 and 22°C | Escherichia coli | |
| 230 | - |
N4-acetyl-2'-deoxycytidine | recombinant C-terminally His6-tagged enzyme, at pH 8.0 and 22°C | Escherichia coli | |
| 400 | - |
N4-acetylcytidine | mutant enzyme H70A, at pH 8.0 and 22°C | Escherichia coli | |
| 1200 | - |
N4-acetylcytidine | mutant enzyme D27A, at pH 8.0 and 22°C | Escherichia coli | |
| 2200 | - |
N4-acetylcytidine | recombinant C-terminally His6-tagged enzyme, at pH 8.0 and 22°C | Escherichia coli | |
| 2500 | - |
N4-acetylcytidine | recombinant N-terminally His6-tagged enzyme, at pH 8.0 and 22°C | Escherichia coli | |
| 2800 | - |
N4-acetylcytidine | recombinant enzyme, at pH 8.0 and 22°C | Escherichia coli | |
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