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Literature summary for 3.6.1.23 extracted from

  • Persson, R.; McGeehan, J.; Wilson, K.S.
    Cloning, expression, purification, and characterisation of the dUTPase encoded by the integrated Bacillus subtilis temperate bacteriophage SPbeta (2005), Protein Expr. Purif., 42, 92-99.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
gene yosS encoded by the Bacillus subtilis temperate bacteriophage SPbeta, DNA and amino acid sequence determination and analysis, functional overexpression in Escherichia coli strain BL21(DE3) Spbetavirus SPbeta

Crystallization (Commentary)

Crystallization (Comment) Organism
purified recombinant free enzyme or recombinant enzyme complexed with substrate dUTP, hanging drop vapour diffusion method, protein and reservoir solution in a 1:1 ratio, 17°C, the reservoir solution contains 0.1 M HEPES, pH 7.5, 10% w/v PEG 8000, and 8% v/v ethylene glycol, 2 days, X-ray diffraction structure determination and analysis at 2.0 A resolution Spbetavirus SPbeta

Inhibitors

Inhibitors Comment Organism Structure
Ba2+
-
Spbetavirus SPbeta
Ca2+
-
Spbetavirus SPbeta
Cd2+
-
Spbetavirus SPbeta

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.0006
-
dUTP pH 7.5, 25°C, recombinant enzyme Spbetavirus SPbeta

Metals/Ions

Metals/Ions Comment Organism Structure
Co2+ activates to 72% of the activity with Mg2+ Spbetavirus SPbeta
Mg2+ required, best divalent cation Spbetavirus SPbeta
Mn2+ activates to 68% of the activity with Mg2+ Spbetavirus SPbeta
additional information no effect by Zn2+ Spbetavirus SPbeta
Ni2+ activates to 49% of the activity with Mg2+ Spbetavirus SPbeta

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
16000
-
3 * 16000, recombinant enzyme, SDS-PAGE Spbetavirus SPbeta
48000
-
recombinant enzyme, gel filtration Spbetavirus SPbeta

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
dUTP + H2O Spbetavirus SPbeta
-
dUMP + diphosphate
-
?

Organism

Organism UniProt Comment Textmining
Spbetavirus SPbeta
-
gene yosS
-

Purification (Commentary)

Purification (Comment) Organism
recombinant enzyme of Bacillus subtilis temperate bacteriophage SPbeta from Escherichia coli strain BL21(DE3) by phosphocellulose chromatography to homogeneity Spbetavirus SPbeta

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
additional information
-
-
Spbetavirus SPbeta
69
-
purified enzyme Spbetavirus SPbeta

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
dUTP + H2O
-
Spbetavirus SPbeta dUMP + diphosphate
-
?

Subunits

Subunits Comment Organism
trimer 3 * 16000, recombinant enzyme, SDS-PAGE Spbetavirus SPbeta

Synonyms

Synonyms Comment Organism
dUTPase
-
Spbetavirus SPbeta

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
25
-
assay at Spbetavirus SPbeta

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
18
-
dUTP pH 7.5, 25°C, recombinant enzyme Spbetavirus SPbeta

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.5
-
assay at Spbetavirus SPbeta

pI Value

Organism Comment pI Value Maximum pI Value
Spbetavirus SPbeta isoelectric focusing
-
6.7