Application | Comment | Organism |
---|---|---|
medicine | cytosine deaminase is used in combination with 5-fluorocytosine as an enzyme-prodrug combination for targeted genetic cancer treatment | Escherichia coli |
Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli BL21(DE3) and BL21-RIL cells | Escherichia coli |
Crystallization (Comment) | Organism |
---|---|
the wild type enzyme in complex with the mechanism-based inhibitors 4-(R)-hydroxyl-3,4-dihydropyrimidine or 5-fluoro-4-(S)-hydroxyl-3,4-dihydropyrimidine and the mutant enzyme V152A/F316C/D317G are crystallized by the sitting drop vapor diffusion method, using 10-15% (w/v) PEG 6K, 200 mM MgCl2 and 100 mM HEPES (pH 7-8) | Escherichia coli |
Protein Variants | Comment | Organism |
---|---|---|
D314E/F316L/D317G | the mutant displays 9% substrate specificity towards cytosine and 1820% substrate specificity towards 5-fluorocytosine compared to the wild type enzyme | Escherichia coli |
V152A/F316C/D317G | the mutant displays 4% substrate specificity towards cytosine and 1920% substrate specificity towards 5-fluorocytosine compared to the wild type enzyme | Escherichia coli |
V315L/F316V/D317G | the mutant displays 6% substrate specificity towards cytosine and 1880% substrate specificity towards 5-fluorocytosine compared to the wild type enzyme | Escherichia coli |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.46 | - |
cytosine | wild type enzyme, pH and temperature not specified in the publication | Escherichia coli | |
1.65 | - |
cytosine | mutant enzyme D314E/F316L/D317G, pH and temperature not specified in the publication | Escherichia coli | |
3.16 | - |
cytosine | mutant enzyme V315L/F316V/D317G, pH and temperature not specified in the publication | Escherichia coli | |
3.76 | - |
5-fluorocytosine | wild type enzyme, pH and temperature not specified in the publication | Escherichia coli | |
4.9 | - |
cytosine | mutant enzyme V152A/F316C/D317G, pH and temperature not specified in the publication | Escherichia coli | |
6.73 | - |
5-fluorocytosine | mutant enzyme D314E/F316L/D317G, pH and temperature not specified in the publication | Escherichia coli | |
7.55 | - |
5-fluorocytosine | mutant enzyme V315L/F316V/D317G, pH and temperature not specified in the publication | Escherichia coli | |
12.69 | - |
5-fluorocytosine | mutant enzyme V152A/F316C/D317G, pH and temperature not specified in the publication | Escherichia coli |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Escherichia coli | - |
- |
- |
Escherichia coli GIA39(DE3) | - |
- |
- |
Purification (Comment) | Organism |
---|---|
Ni-NTA column chromatography | Escherichia coli |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
5-fluorocytosine + H2O | - |
Escherichia coli | 5-fluorouracil + NH3 | - |
? | |
5-fluorocytosine + H2O | - |
Escherichia coli GIA39(DE3) | 5-fluorouracil + NH3 | - |
? | |
cytosine + H2O | - |
Escherichia coli | uracil + NH3 | - |
? | |
cytosine + H2O | - |
Escherichia coli GIA39(DE3) | uracil + NH3 | - |
? |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
1.69 | - |
cytosine | mutant enzyme V152A/F316C/D317G, pH and temperature not specified in the publication | Escherichia coli | |
2.13 | - |
cytosine | mutant enzyme D314E/F316L/D317G, pH and temperature not specified in the publication | Escherichia coli | |
2.98 | - |
cytosine | mutant enzyme V315L/F316V/D317G, pH and temperature not specified in the publication | Escherichia coli | |
19.71 | - |
5-fluorocytosine | wild type enzyme, pH and temperature not specified in the publication | Escherichia coli | |
49.68 | - |
cytosine | wild type enzyme, pH and temperature not specified in the publication | Escherichia coli | |
83.68 | - |
5-fluorocytosine | mutant enzyme D314E/F316L/D317G, pH and temperature not specified in the publication | Escherichia coli | |
101.7 | - |
5-fluorocytosine | mutant enzyme V152A/F316C/D317G, pH and temperature not specified in the publication | Escherichia coli | |
115.2 | - |
5-fluorocytosine | mutant enzyme V315L/F316V/D317G, pH and temperature not specified in the publication | Escherichia coli |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.35 | - |
cytosine | mutant enzyme V152A/F316C/D317G, pH and temperature not specified in the publication | Escherichia coli | |
0.94 | - |
cytosine | mutant enzyme V315L/F316V/D317G, pH and temperature not specified in the publication | Escherichia coli | |
1.29 | - |
cytosine | mutant enzyme D314E/F316L/D317G, pH and temperature not specified in the publication | Escherichia coli | |
5.24 | - |
5-fluorocytosine | wild type enzyme, pH and temperature not specified in the publication | Escherichia coli | |
8.02 | - |
5-fluorocytosine | mutant enzyme V152A/F316C/D317G, pH and temperature not specified in the publication | Escherichia coli | |
12.44 | - |
5-fluorocytosine | mutant enzyme D314E/F316L/D317G, pH and temperature not specified in the publication | Escherichia coli | |
15.27 | - |
5-fluorocytosine | mutant enzyme V315L/F316V/D317G, pH and temperature not specified in the publication | Escherichia coli | |
106.8 | - |
cytosine | wild type enzyme, pH and temperature not specified in the publication | Escherichia coli |