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Literature summary for 3.5.4.1 extracted from

  • Mahan, S.D.; Ireton, G.C.; Knoeber, C.; Stoddard, B.L.; Black, M.E.
    Random mutagenesis and selection of Escherichia coli cytosine deaminase for cancer gene therapy (2004), Protein Eng. Des. Sel., 17, 625-633.
    View publication on PubMed
Search for more literature for 3.5.4.1

Crystallization (Commentary)

Crystallization (Comment) Organism
wild-type enzyme and mutants D314G, D314A, D314S, free and in complex with mechanism-based inhibitor 5-fluoro-4-(S)-hydroxyl-3,4-dihydropyrimidine Escherichia coli

Protein Variants

Protein Variants Comment Organism
D314A impaired catalytic efficiency for cytosine, improved catalytic efficiency for 5-fluorocytosine Escherichia coli
D314G impaired catalytic efficiency Escherichia coli
D314S impaired catalytic efficiency Escherichia coli

Inhibitors

Inhibitors Comment Organism Structure
5-fluoro-4-(S)-hydroxyl-3,4-dihydropyrimidine mechanism-based inhibitor Escherichia coli

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.2
-
 cytosine wild-type, pH 7.5, 37°C Escherichia coli
0.3
-
 cytosine mutant D314G, pH 7.5, 37°C Escherichia coli
1
-
 cytosine mutant D314S, pH 7.5, 37°C Escherichia coli
2.2
-
 cytosine mutant D314A, pH 7.5, 37°C Escherichia coli
2.8
-
 5-fluorocytosine mutant D314A, pH 7.5, 37°C Escherichia coli
3.3
-
 5-fluorocytosine wild-type, pH 7.5, 37°C Escherichia coli
6.6
-
 5-fluorocytosine mutant D314S, pH 7.5, 37°C Escherichia coli
10.9
-
 5-fluorocytosine mutant D314G, pH 7.5, 37°C Escherichia coli

Organism

Organism UniProt Comment Textmining
Escherichia coli
-
-
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
5-fluorocytosine + H2O
-
 Escherichia coli 5-fluorouracil + NH3
-
 ?
cytosine + H2O
-
 Escherichia coli uracil + NH3
-
 ?

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
36.8
-
 cytosine mutant D314G, pH 7.5, 37°C Escherichia coli
75.6
-
 5-fluorocytosine wild-type, pH 7.5, 37°C Escherichia coli
97.7
-
 5-fluorocytosine mutant D314G, pH 7.5, 37°C Escherichia coli
104
-
 cytosine mutant D314A, pH 7.5, 37°C Escherichia coli
137
-
 5-fluorocytosine mutant D314A, pH 7.5, 37°C Escherichia coli
165
-
 cytosine wild-type, pH 7.5, 37°C Escherichia coli
233
-
 5-fluorocytosine mutant D314S, pH 7.5, 37°C Escherichia coli
279
-
 cytosine mutant D314S, pH 7.5, 37°C Escherichia coli