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Literature summary extracted from

  • Andersen, R.B.; Neuhard, J.
    Deoxynucleoside kinases encoded by the yaaG and yaaF genes of Bacillus subtilis. Substrate specificity and kinetic analysis of deoxyguanosine kinase with UTP as the preferred phosphate donor (2001), J. Biol. Chem., 276, 5518-5524.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.7.1.76 gene yaaF of the yaaG/yaaF operon, DNA and amino acid sequence determination and analysis, overexpression in Escherichia coli Bacillus subtilis
2.7.1.113 expression in Escherichia coli Bacillus subtilis
2.7.1.113 gene yaaG of the yaaG/yaaF operon, DNA and amino acid sequence determination and analysis, overexpression in Escherichia coli Bacillus subtilis

General Stability

EC Number General Stability Organism
2.7.1.113 bovine serum albumin stabilizes the enzyme in solution Bacillus subtilis
2.7.1.113 highly unstable at 0°C in diluted form, no loss of activity in the presence of bovine serum albumin Bacillus subtilis

Inhibitors

EC Number Inhibitors Comment Organism Structure
2.7.1.113 dGMP noncompetitive versus UTP and 2'-deoxyguanosine Bacillus subtilis
2.7.1.113 additional information no substrate inhibition by CTP, ATP, and GTP Bacillus subtilis
2.7.1.113 UDP competitive versus UTP, noncompetitive versus 2'-deoxyguanosine Bacillus subtilis
2.7.1.113 UTP substrate inhibition, overview Bacillus subtilis

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.7.1.113 0.0006
-
 2'-deoxyguanosine pH 8.5, 37°C, cosubstrate UTP Bacillus subtilis
2.7.1.113 0.0006
-
 2'-deoxyguanosine pH 8.5, 37°C, recombinant enzyme, with UTP as donor substrate Bacillus subtilis
2.7.1.113 0.0017
-
 2'-deoxyguanosine pH 8.5, 37°C, recombinant enzyme, with CTP as donor substrate Bacillus subtilis
2.7.1.113 0.006
-
 UTP pH 8.5, 37°C Bacillus subtilis
2.7.1.113 0.006
-
 UTP pH 8.5, 37°C, recombinant enzyme, with 2'-deoxyguanosine as acceptor substrate Bacillus subtilis
2.7.1.113 0.0065
-
 2'-deoxyguanosine pH 8.5, 37°C, cosubstrate ATP Bacillus subtilis
2.7.1.113 0.0065
-
 2'-deoxyguanosine pH 8.5, 37°C, recombinant enzyme, with ATP as donor substrate Bacillus subtilis
2.7.1.113 0.01
-
 2'-deoxyguanosine pH 8.5, 37°C, cosubstrate GTP Bacillus subtilis
2.7.1.113 0.0104
-
 2'-deoxyguanosine pH 8.5, 37°C, recombinant enzyme, with GTP as donor substrate Bacillus subtilis
2.7.1.113 0.017
-
 2'-deoxyguanosine pH 8.5, 37°C, cosubstrate CTP Bacillus subtilis
2.7.1.113 0.035
-
 CTP pH 8.5, 37°C Bacillus subtilis
2.7.1.113 0.035
-
 CTP pH 8.5, 37°C, recombinant enzyme, with 2'-deoxyguanosine as acceptor substrate Bacillus subtilis
2.7.1.113 0.036
-
 ATP pH 8.5, 37°C Bacillus subtilis
2.7.1.113 0.036
-
 ATP pH 8.5, 37°C, recombinant enzyme, with 2'-deoxyguanosine as acceptor substrate Bacillus subtilis
2.7.1.113 0.046
-
 GTP pH 8.5, 37°C Bacillus subtilis
2.7.1.113 0.046
-
 GTP pH 8.5, 37°C, recombinant enzyme, with 2'-deoxyguanosine as acceptor substrate Bacillus subtilis

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
2.7.1.76 Mg2+ required for activity Bacillus subtilis
2.7.1.113 Mg2+ required for activity Bacillus subtilis
2.7.1.113 Mg2+ MgNTP2- is most probably the true phosphate donor Bacillus subtilis
2.7.1.113 Mg2+ required for activity, no inhibition by excess Mg2+, at 5 mM Bacillus subtilis

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
2.7.1.113 24147
-
 2 * 24147, electrospray mass spectrometry Bacillus subtilis
2.7.1.113 24147
-
 2 * 24147, recombinant enzyme, electrospray mass spectrometry Bacillus subtilis
2.7.1.113 49000
-
 recombinant enzyme, gel filtration Bacillus subtilis
2.7.1.113 49000
-
 recombinant deoxyguanosine kinase, gel filtration Bacillus subtilis

Organism

EC Number Organism UniProt Comment Textmining
2.7.1.76 Bacillus subtilis
-
 gene yaaF, yaaF encodes an enzyme with specificity for deoxyadenosine and deoxycytidine
-
2.7.1.113 Bacillus subtilis
-
-
-
2.7.1.113 Bacillus subtilis
-
 gene yaaG
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.7.1.113 recombinant deoxyguanosine kinase, BlueA dye affinity, phenyl-Sepharose Bacillus subtilis
2.7.1.113 recombinant enzyme from Escherichia coli by affinity and phenyl resin chromatography to homogeneity Bacillus subtilis

Reaction

EC Number Reaction Comment Organism Reaction ID
2.7.1.113 ATP + deoxyguanosine = ADP + dGMP sequential Bi Bi mechanism Bacillus subtilis
a

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
2.7.1.76 2.3
-
 dAdo/dCyd kinase in crude recombinant Escherichia coli cell extract, substrate 2'-deoxyadenosine or 2'-deoxycytidine Bacillus subtilis
2.7.1.113 0.54
-
 enzyme in crude recombinant Escherichia coli cell extract, substrate 2'-deoxyguanosine Bacillus subtilis
2.7.1.113 2.3
-
 dAdo/dCyd kinase in crude recombinant Escherichia coli cell extract, substrate 2'-deoxyadenosine or 2'-deoxycytidine Bacillus subtilis
2.7.1.113 8.1
-
 recombinant deoxyguanosine kinase Bacillus subtilis
2.7.1.113 8.1
-
 purified recombinant enzyme, substrate 2'-deoxyguanosine Bacillus subtilis

Storage Stability

EC Number Storage Stability Organism
2.7.1.113 -20°C, more than 1 mg/ml protein, 50% glycerol, 1 year, no loss of activity Bacillus subtilis
2.7.1.113 -20°C, purified recombinant enzyme at 1 mg/ml, Tris-HCl, pH 7.8, 50% v/v glycerol, at least 1 year without loss of activity Bacillus subtilis
2.7.1.113 0°C, the diluted enzyme is highly unstable, but in presence of 1 mg/ml bovine serum albumin, the diluted enzyme is stable for months Bacillus subtilis

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.7.1.76 GTP + 2'-deoxyadenosine
-
 Bacillus subtilis GDP + 2'-deoxyadenosine 5'-phosphate
-
 ?
2.7.1.76 GTP + 2'-deoxycytidine
-
 Bacillus subtilis GDP + 2'-deoxycytidine 5'-phosphate
-
 ?
2.7.1.76 additional information substrate specificity, enzyme is specific for deoxyadenosine and deoxycytidine Bacillus subtilis ?
-
 ?
2.7.1.113 ATP + 2'-deoxyguanosine
-
 Bacillus subtilis ADP + 2'-deoxyguanosine 5'-phosphate
-
 ?
2.7.1.113 ATP + deoxyadenosine 3% of activity with deoxyguanosine Bacillus subtilis ADP + dAMP
-
 ?
2.7.1.113 ATP + deoxycytidine recombinant deoxyguanosine kinase, 3% of activity with deoxyguanosine Bacillus subtilis ADP + dCMP
-
 ?
2.7.1.113 ATP + deoxyinosine 63% of activity with deoxyguanosine Bacillus subtilis ADP + dIMP
-
 ?
2.7.1.113 ATP + guanosine recombinant deoxyguanosine kinase, 7% of activity with deoxyguanosine Bacillus subtilis ADP + GMP
-
 ?
2.7.1.113 CTP + 2'-deoxyguanosine
-
 Bacillus subtilis CDP + 2'-deoxyguanosine 5'-phosphate
-
 ?
2.7.1.113 GTP + 2'-deoxyadenosine very low activity Bacillus subtilis GDP + 2'-deoxyadenosine 5'-phosphate
-
 ?
2.7.1.113 GTP + 2'-deoxycytidine very low activity Bacillus subtilis GDP + 2'-deoxycytidine 5'-phosphate
-
 ?
2.7.1.113 GTP + 2'-deoxyguanosine
-
 Bacillus subtilis GDP + 2'-deoxyguanosine 5'-phosphate
-
 ?
2.7.1.113 GTP + 2'-deoxyinosine
-
 Bacillus subtilis GDP + 2'-deoxyinosine 5'-phosphate
-
 ?
2.7.1.113 GTP + guanosine very low activity Bacillus subtilis GDP + guanosine 5'-phosphate
-
 ?
2.7.1.113 additional information substrate specificity, the enzyme utilizes all naturally occurring ribo-and deoxyribonucleosides triphosphates, with the exception of dGTP, as phosphate donors, and shows poor activity with thymidine, inosine, adenosine, 2,3-dideoxyguanosine, 2,3-dideoxyinosine, 2,3-dideoxyadenosine, and hypoxanthine arabinoside as acceptor substrates Bacillus subtilis ?
-
 ?
2.7.1.113 UTP + 2'-deoxyguanosine preferred substrates Bacillus subtilis UDP + 2'-deoxyguanosine 5'-phosphate
-
 ?

Subunits

EC Number Subunits Comment Organism
2.7.1.113 dimer 2 * 24147, electrospray mass spectrometry Bacillus subtilis
2.7.1.113 dimer 2 * 24147, recombinant enzyme, electrospray mass spectrometry Bacillus subtilis

Synonyms

EC Number Synonyms Comment Organism
2.7.1.76 dAdo/dCyd kinase
-
 Bacillus subtilis
2.7.1.113 deoxyguanosine kinase
-
 Bacillus subtilis
2.7.1.113 DGK
-
 Bacillus subtilis

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
2.7.1.76 37
-
 assay at Bacillus subtilis
2.7.1.113 37
-
 assay at Bacillus subtilis

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
2.7.1.113 1.41
-
 deoxyguanosine pH 8.5, 37°C, cosubstrate UTP Bacillus subtilis
2.7.1.113 1.41
-
 UTP pH 8.5, 37°C, recombinant enzyme, with 2'-deoxyguanosine as acceptor substrate Bacillus subtilis
2.7.1.113 1.41
-
 2'deoxyguanosine pH 8.5, 37°C, recombinant enzyme, with UTP as donor substrate Bacillus subtilis
2.7.1.113 1.53
-
 deoxyguanosine pH 8.5, 37°C, cosubstrate ATP Bacillus subtilis
2.7.1.113 1.53
-
 ATP pH 8.5, 37°C, recombinant enzyme, with 2'-deoxyguanosine as acceptor substrate Bacillus subtilis
2.7.1.113 1.53
-
 2'deoxyguanosine pH 8.5, 37°C, recombinant enzyme, with ATP as donor substrate Bacillus subtilis
2.7.1.113 1.69
-
 deoxyguanosine pH 8.5, 37°C, cosubstrate GTP Bacillus subtilis
2.7.1.113 1.69
-
 GTP pH 8.5, 37°C, recombinant enzyme, with 2'-deoxyguanosine as acceptor substrate Bacillus subtilis
2.7.1.113 1.69
-
 2'deoxyguanosine pH 8.5, 37°C, recombinant enzyme, with GTP as donor substrate Bacillus subtilis
2.7.1.113 2.25
-
 deoxyguanosine pH 8.5, 37°C, cosubstrate CTP Bacillus subtilis
2.7.1.113 2.25
-
 CTP pH 8.5, 37°C, recombinant enzyme, with 2'-deoxyguanosine as acceptor substrate Bacillus subtilis
2.7.1.113 2.25
-
 2'deoxyguanosine pH 8.5, 37°C, recombinant enzyme, with CTP as donor substrate Bacillus subtilis

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
2.7.1.76 8.5
-
 assay at Bacillus subtilis
2.7.1.113 9
-
-
 Bacillus subtilis
2.7.1.113 9
-
 broad optimum Bacillus subtilis

pH Range

EC Number pH Minimum pH Maximum Comment Organism
2.7.1.113 5.6 9 more than 80% of maximal activity at pH 7.5 and pH 11.5, respectively, 60% of maximal activity at pH 6.0, complete loss of activity below pH 5.6 Bacillus subtilis
2.7.1.113 6 11.5 over 80% of maximal activity at pH 7.5 and pH 11.5, about 60% of maximal activity at pH 6.0, enzyme is completely inactive below pH 5.6 Bacillus subtilis

Cofactor

EC Number Cofactor Comment Organism Structure
2.7.1.76 GTP
-
 Bacillus subtilis
2.7.1.113 ATP
-
 Bacillus subtilis
2.7.1.113 CTP
-
 Bacillus subtilis
2.7.1.113 GTP
-
 Bacillus subtilis
2.7.1.113 additional information the enzyme utilizes all naturally occurring ribo-and deoxyribonucleosides triphosphates, with the exception of dGTP, as phosphate donors Bacillus subtilis
2.7.1.113 UTP
-
 Bacillus subtilis

Ki Value [mM]

EC Number Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
2.7.1.113 additional information
-
 additional information product inhibition of the enzyme, overview Bacillus subtilis
2.7.1.113 0.063
-
 UDP pH 8.5, 37°C, cosubstrate UTP Bacillus subtilis