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Literature summary extracted from

  • Mahan, S.D.; Ireton, G.C.; Stoddard, B.L.; Black, M.E.
    Alanine-scanning mutagenesis reveals a cytosine deaminase mutant with altered substrate preference (2004), Biochemistry, 43, 8957-8964.
    View publication on PubMed

Application

EC Number Application Comment Organism
3.5.4.1 medicine mutant enzyme with improved activity towards cancer prodrug 5-fluorouracil for suicide gene therapy Escherichia coli

Protein Variants

EC Number Protein Variants Comment Organism
3.5.4.1 D314A 11fold increase in KM-value with cytosine, decrease in KM-value with 5-fluorocytosine Escherichia coli
3.5.4.1 F310A no activity with cytosine Escherichia coli
3.5.4.1 F316A KM-value with cytosine similar to wild-type, decrease in KM-value with 5-fluorocytosine Escherichia coli
3.5.4.1 G311A no activity with cytosine Escherichia coli
3.5.4.1 H312A 3fold increase in KM-value with cytosine Escherichia coli
3.5.4.1 P318A 4fold increase in KM-value with cytosine Escherichia coli
3.5.4.1 V315A no activity with cytosine Escherichia coli

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.5.4.1 0.2
-
 cytosine pH 7.5, 37°C, wild-type Escherichia coli
3.5.4.1 0.2
-
 cytosine pH 7.5, 37°C, mutant F316A Escherichia coli
3.5.4.1 0.7
-
 cytosine pH 7.5, 37°C, mutant H312A Escherichia coli
3.5.4.1 0.8
-
 cytosine pH 7.5, 37°C, mutant P318A Escherichia coli
3.5.4.1 2.1
-
 5-fluorocytosine pH 7.5, 37°C, mutant F316A Escherichia coli
3.5.4.1 2.2
-
 cytosine pH 7.5, 37°C, mutant D314A Escherichia coli
3.5.4.1 2.8
-
 5-fluorocytosine pH 7.5, 37°C, mutant D314A Escherichia coli
3.5.4.1 3.3
-
 5-fluorocytosine pH 7.5, 37°C, wild-type Escherichia coli

Organism

EC Number Organism UniProt Comment Textmining
3.5.4.1 Escherichia coli
-
-
-

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.5.4.1 5-fluorocytosine + H2O
-
 Escherichia coli 5-fluorouracil + NH3
-
 ?
3.5.4.1 cytosine + H2O
-
 Escherichia coli uracil + NH3
-
 ?

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
3.5.4.1 74.3
-
 5-fluorocytosine pH 7.5, 37°C, mutant F316A Escherichia coli
3.5.4.1 75.6
-
 5-fluorocytosine pH 7.5, 37°C, wild-type Escherichia coli
3.5.4.1 104
-
 cytosine pH 7.5, 37°C, mutant D314A Escherichia coli
3.5.4.1 137.4
-
 5-fluorocytosine pH 7.5, 37°C, mutant D314A Escherichia coli
3.5.4.1 165
-
 cytosine pH 7.5, 37°C, wild-type Escherichia coli
3.5.4.1 189
-
 cytosine pH 7.5, 37°C, mutant H312A Escherichia coli
3.5.4.1 1030
-
 cytosine pH 7.5, 37°C, mutant P318A Escherichia coli
3.5.4.1 2286
-
 cytosine pH 7.5, 37°C, mutant F316A Escherichia coli