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Literature summary 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)

EC Number Cloned (Comment) Organism
3.6.1.23 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)

EC Number Crystallization (Comment) Organism
3.6.1.23 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

EC Number Inhibitors Comment Organism Structure
3.6.1.23 Ba2+
-
 Spbetavirus SPbeta
3.6.1.23 Ca2+
-
 Spbetavirus SPbeta
3.6.1.23 Cd2+
-
 Spbetavirus SPbeta

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.6.1.23 0.0006
-
 dUTP pH 7.5, 25°C, recombinant enzyme Spbetavirus SPbeta

Metals/Ions

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

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
3.6.1.23 16000
-
 3 * 16000, recombinant enzyme, SDS-PAGE Spbetavirus SPbeta
3.6.1.23 48000
-
 recombinant enzyme, gel filtration Spbetavirus SPbeta

Natural Substrates/ Products (Substrates)

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

Organism

EC Number Organism UniProt Comment Textmining
3.6.1.23 Spbetavirus SPbeta
-
 gene yosS
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.6.1.23 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]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
3.6.1.23 additional information
-
-
 Spbetavirus SPbeta
3.6.1.23 69
-
 purified enzyme Spbetavirus SPbeta

Substrates and Products (Substrate)

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

Subunits

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

Synonyms

EC Number Synonyms Comment Organism
3.6.1.23 dUTPase
-
 Spbetavirus SPbeta

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
3.6.1.23 25
-
 assay at Spbetavirus SPbeta

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
3.6.1.23 18
-
 dUTP pH 7.5, 25°C, recombinant enzyme Spbetavirus SPbeta

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
3.6.1.23 7.5
-
 assay at Spbetavirus SPbeta

pI Value

EC Number Organism Comment pI Value Maximum pI Value
3.6.1.23 Spbetavirus SPbeta isoelectric focusing
-
 6.7