BRENDA - Enzyme Database show

In vivo selection of active deoxyribonucleoside kinase by a mutagenic nucleoside analog

Kamiya, H.; Ito, M.; Nishi, K.; Harashima, H.; J. Biotechnol. 228, 52-57 (2016)

Data extracted from this reference:

Cloned(Commentary)
EC Number
Commentary
Organism
2.7.1.145
gene dnk, isolated from a phage library, DNA and amino acid sequence determination and analysis, recombinant expression of randomly mutated genes for deoxyribonucleoside kinase in Escherichia coli strain BL21, recombinant expression of catalytically active His10-tagged mutant 4 enzyme in Spodoptera frugiperda SF9 cells
Drosophila melanogaster
Inhibitors
EC Number
Inhibitors
Commentary
Organism
Structure
2.7.1.145
EDTA
-
Drosophila melanogaster
Metals/Ions
EC Number
Metals/Ions
Commentary
Organism
Structure
2.7.1.145
Mg2+
required
Drosophila melanogaster
Natural Substrates/ Products (Substrates)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
2.7.1.145
ATP + 2'-deoxyribonucleoside
Drosophila melanogaster
-
ADP + 2'-deoxyribonucleoside 5'-phosphate
-
-
?
Organism
EC Number
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
2.7.1.145
Drosophila melanogaster
Q9XZT6
gene dnk
-
Purification (Commentary)
EC Number
Commentary
Organism
2.7.1.145
recombinant His10-tagged enzyme from Spodoptera frugiperda SF9 cells by nickel affinity chromatography and gel filtration
Drosophila melanogaster
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
2.7.1.145
ATP + 2'-deoxycytidine
-
738739
Drosophila melanogaster
ADP + 2'-deoxycytidine 5'-phosphate
-
-
-
?
2.7.1.145
ATP + 2'-deoxyribonucleoside
-
738739
Drosophila melanogaster
ADP + 2'-deoxyribonucleoside 5'-phosphate
-
-
-
?
2.7.1.145
ATP + 2-hydroxy-2'-deoxyadenosine
preferred substrate for enzyme mutant 4
738739
Drosophila melanogaster
ADP + 2-hydroxy-2'-deoxyadenosine 5'-monophosphate
-
-
-
?
Temperature Optimum [°C]
EC Number
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
2.7.1.145
37
-
assay at
Drosophila melanogaster
pH Optimum
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
2.7.1.145
8
-
assay at
Drosophila melanogaster
Cofactor
EC Number
Cofactor
Commentary
Organism
Structure
2.7.1.145
ATP
-
Drosophila melanogaster
Cloned(Commentary) (protein specific)
EC Number
Commentary
Organism
2.7.1.145
gene dnk, isolated from a phage library, DNA and amino acid sequence determination and analysis, recombinant expression of randomly mutated genes for deoxyribonucleoside kinase in Escherichia coli strain BL21, recombinant expression of catalytically active His10-tagged mutant 4 enzyme in Spodoptera frugiperda SF9 cells
Drosophila melanogaster
Cofactor (protein specific)
EC Number
Cofactor
Commentary
Organism
Structure
2.7.1.145
ATP
-
Drosophila melanogaster
Inhibitors (protein specific)
EC Number
Inhibitors
Commentary
Organism
Structure
2.7.1.145
EDTA
-
Drosophila melanogaster
Metals/Ions (protein specific)
EC Number
Metals/Ions
Commentary
Organism
Structure
2.7.1.145
Mg2+
required
Drosophila melanogaster
Natural Substrates/ Products (Substrates) (protein specific)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
2.7.1.145
ATP + 2'-deoxyribonucleoside
Drosophila melanogaster
-
ADP + 2'-deoxyribonucleoside 5'-phosphate
-
-
?
Purification (Commentary) (protein specific)
EC Number
Commentary
Organism
2.7.1.145
recombinant His10-tagged enzyme from Spodoptera frugiperda SF9 cells by nickel affinity chromatography and gel filtration
Drosophila melanogaster
Substrates and Products (Substrate) (protein specific)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
2.7.1.145
ATP + 2'-deoxycytidine
-
738739
Drosophila melanogaster
ADP + 2'-deoxycytidine 5'-phosphate
-
-
-
?
2.7.1.145
ATP + 2'-deoxyribonucleoside
-
738739
Drosophila melanogaster
ADP + 2'-deoxyribonucleoside 5'-phosphate
-
-
-
?
2.7.1.145
ATP + 2-hydroxy-2'-deoxyadenosine
preferred substrate for enzyme mutant 4
738739
Drosophila melanogaster
ADP + 2-hydroxy-2'-deoxyadenosine 5'-monophosphate
-
-
-
?
Temperature Optimum [°C] (protein specific)
EC Number
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
2.7.1.145
37
-
assay at
Drosophila melanogaster
pH Optimum (protein specific)
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
2.7.1.145
8
-
assay at
Drosophila melanogaster
General Information
EC Number
General Information
Commentary
Organism
2.7.1.145
additional information
enzyme identification using a mutagenic nucleoside analogue 2-hydroxy-2'-deoxyadenosine 5'-monophosphate, overview. Method development and evaluation for in vivo evolution selection method for active deoxyribonucleoside kinase proteins, including selection of antibiotic-resistant colonies resulting frommutations by phosphorylated 2-hydroxy-deoxyadenosine and the subsequent isolation of the plasmid DNAs. Fifteen Dm-dNK mutants selected after the seventh and eighth evolutioncycles are actually active in vivo, one shows activity similar to the wild-typ enzyme
Drosophila melanogaster
General Information (protein specific)
EC Number
General Information
Commentary
Organism
2.7.1.145
additional information
enzyme identification using a mutagenic nucleoside analogue 2-hydroxy-2'-deoxyadenosine 5'-monophosphate, overview. Method development and evaluation for in vivo evolution selection method for active deoxyribonucleoside kinase proteins, including selection of antibiotic-resistant colonies resulting frommutations by phosphorylated 2-hydroxy-deoxyadenosine and the subsequent isolation of the plasmid DNAs. Fifteen Dm-dNK mutants selected after the seventh and eighth evolutioncycles are actually active in vivo, one shows activity similar to the wild-typ enzyme
Drosophila melanogaster