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Literature summary for 3.5.4.1 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
Search for more literature for 3.5.4.1

Application

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

Protein Variants

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

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.2
-
 cytosine pH 7.5, 37°C, wild-type Escherichia coli
0.2
-
 cytosine pH 7.5, 37°C, mutant F316A Escherichia coli
0.7
-
 cytosine pH 7.5, 37°C, mutant H312A Escherichia coli
0.8
-
 cytosine pH 7.5, 37°C, mutant P318A Escherichia coli
2.1
-
 5-fluorocytosine pH 7.5, 37°C, mutant F316A Escherichia coli
2.2
-
 cytosine pH 7.5, 37°C, mutant D314A Escherichia coli
2.8
-
 5-fluorocytosine pH 7.5, 37°C, mutant D314A Escherichia coli
3.3
-
 5-fluorocytosine pH 7.5, 37°C, wild-type 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
74.3
-
 5-fluorocytosine pH 7.5, 37°C, mutant F316A Escherichia coli
75.6
-
 5-fluorocytosine pH 7.5, 37°C, wild-type Escherichia coli
104
-
 cytosine pH 7.5, 37°C, mutant D314A Escherichia coli
137.4
-
 5-fluorocytosine pH 7.5, 37°C, mutant D314A Escherichia coli
165
-
 cytosine pH 7.5, 37°C, wild-type Escherichia coli
189
-
 cytosine pH 7.5, 37°C, mutant H312A Escherichia coli
1030
-
 cytosine pH 7.5, 37°C, mutant P318A Escherichia coli
2286
-
 cytosine pH 7.5, 37°C, mutant F316A Escherichia coli