BRENDA - Enzyme Database show

Characterization of N-deoxyribosyltransferase from Lactococcus lactis subsp. lactis

Miyamoto, Y.; Masaki, T.; Chohnan, S.; Biochim. Biophys. Acta 1774, 1323-1330 (2007)

Data extracted from this reference:

Activating Compound
EC Number
Activating Compound
Commentary
Organism
Structure
2.4.2.6
K+
K+-activated type II enzyme: binding of K+ is required to enhance activity through induced conformational change
Lactococcus lactis
2.4.2.6
additional information
does not require phosphate derivatives such as ATP, thiamine diphosphate, pyridoxal phosphate to complete 2-deoxyribose transfer
Lactococcus lactis
Cloned(Commentary)
EC Number
Commentary
Organism
2.4.2.6
in pET-22b(+) for expression in Escherichia coli BL21(DE3); in pGEM-T for cloning
Lactococcus lactis
Inhibitors
EC Number
Inhibitors
Commentary
Organism
Structure
2.4.2.6
Ca2+
inhibits base exchange activity at 1 mM
Lactococcus lactis
2.4.2.6
CO2
reduces base exchange activity by 87-99% at 1 mM
Lactococcus lactis
2.4.2.6
Cu2+
reduces base exchange activity by 87-99% at 1 mM
Lactococcus lactis
2.4.2.6
Mg2+
inhibits base exchange activity at 1mM
Lactococcus lactis
2.4.2.6
Mn2+
reduces base exchange activity by 87-99% at 1 mM
Lactococcus lactis
2.4.2.6
Zn2+
reduces base exchange activity by 87-99% at 1 mM
Lactococcus lactis
Metals/Ions
EC Number
Metals/Ions
Commentary
Organism
Structure
2.4.2.6
K+
enhances base exchange activity and thermostability, 1.66-fold activation by 1 mM KCl (reaches plateau at 20 mM)
Lactococcus lactis
2.4.2.6
additional information
enzymatic activity depends on monovalent cations as cofactors rather than catalysts; K+ and NH4+ have no synergistic effects; Li+ and Na+ do not activate
Lactococcus lactis
2.4.2.6
NH4+
enhances base exchange activity and thermostability, 1.43-fold activation by 1 mM (NH4)2SO4 (reaches plateau at 20 mM)
Lactococcus lactis
2.4.2.6
Rb+
enhances base exchange activity, 1.51-fold activation by 1 mM RbCl
Lactococcus lactis
Molecular Weight [Da]
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
2.4.2.6
17250
-
4 * 17250, elution as single peak from the preparative gel filtration column
Lactococcus lactis
2.4.2.6
18290
-
theoretical molecular weight
Lactococcus lactis
2.4.2.6
25500
-
according to migration in SDS-12.5% polyacrylamide gel electrophoresis
Lactococcus lactis
2.4.2.6
69000
-
gel filtration chromatography
Lactococcus lactis
Natural Substrates/ Products (Substrates)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
2.4.2.6
additional information
Lactococcus lactis
preferably purine 2-deoxyribonucleoside synthesis compared to pyrimidine deoxyribonucleoside synthesis although both are catalysed
?
-
-
-
2.4.2.6
purine base + 2-deoxy-D-ribosyl-base
Lactococcus lactis
-
purine deoxyribonucleoside + base
preferred over pyrimidine deoxyribonucleoside synthesis
-
r
2.4.2.6
pyrimidine base + 2-deoxy-D-ribosyl-base
Lactococcus lactis
-
pyrimidine deoxyribonucleoside + base
-
-
r
Organism
EC Number
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
2.4.2.6
Lactococcus lactis
-
subsp. lactis JCM5808
-
Purification (Commentary)
EC Number
Commentary
Organism
2.4.2.6
preclearing of bacterial lysate with 40% ammonium sulfate and subsequent precipitation of proteins with 90% ammonium sulfate, gel filtration of dissolved protein on Sephacryl S-200 column, combination of fractions with N-deoxyribosyltransferase activity, precipitation of proteins with 90% ammonium sulfate, dialysis of dissolved proteins and concentration by ultrafiltration, second gel filtration on Sephacryl S-200 column, combination of active fractions and concentration by ultrafiltration, recovery of 70.4% of activity, storage at -20°C in 50% glycerol
Lactococcus lactis
Specific Activity [micromol/min/mg]
EC Number
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
2.4.2.6
additional information
-
0.9% in presence of 1 mM ZnCl2; 102% in presence of 1 mM NaCl; 105% in presence of 1 mM CsCl; 106% in presence of 1 mM NaSO4; 12.9% in presence of 1 mM CoCl2; 143% in presence of 1 mM NH4Cl; 151% in presence of 1 mM RbCl; 163% in presence of 1 mM (NH4)2SO4; 165% in presence of 1 mM K2SO4; 166% in presence of 1 mM KCl; 166% in presence of 1 mM (NH4)2SO4 and 1 mM KCl; 192% in presence of 10 mM KCl; 194% in presence of 10 mM (NH4)2SO4; 3.2% in presence of 1 mM CuSO4; 51.8% in presence of 1 mM CaCl2; 66.3% in presence of 1 mM MgCl2; 66% in presence of 1 mM MgSO4; 68.3% in presence of 1 mM FeCl2; 97.7% in presence of 1 mM LiCl
Lactococcus lactis
2.4.2.6
6.67
-
equals 100%, purified protein, in presence of 0.5 mg/ml bovine serum albumin, pH 7.5, 10 min, 25°C
Lactococcus lactis
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
2.4.2.6
adenine + deoxycytidine
64.0% conversion after 4 h at standard reaction conditions: 2 mM each substrate, 0.013 U NTD, pH 7.5, 25°C, 40 mM (NH4)2SO4, 0.5 mg/ml bovine serum albumin
691053
Lactococcus lactis
deoxyadenosine + cytosine
-
-
-
r
2.4.2.6
adenine + deoxyguanosine
46.6% conversion after 4 h at standard reaction conditions: 2 mM each substrate, 0.013 U NTD, pH 7.5, 25°C, 40 mM (NH4)2SO4, 0.5 mg/ml bovine serum albumin
691053
Lactococcus lactis
deoxyadenosine + guanine
-
-
-
r
2.4.2.6
adenine + deoxyinosine
57.3% conversion after 4 h at standard reaction conditions: 2 mM each substrate, 0.013 U NTD, pH 7.5, 25°C, 40 mM (NH4)2SO4, 0.5 mg/ml bovine serum albumin
691053
Lactococcus lactis
deoxyadenosine + hypoxanthine
-
-
-
r
2.4.2.6
adenine + deoxyuridine
75.4% conversion after 4 h at standard reaction conditions: 2 mM each substrate, 0.013 U NTD, pH 7.5, 25°C, 40 mM (NH4)2SO4, 0.5 mg/ml bovine serum albumin
691053
Lactococcus lactis
deoxyadenosine + uracil
-
-
-
r
2.4.2.6
adenine + thymidine
74.6% conversion after 4 h at standard reaction conditions: 2 mM each substrate, 0.013 U NTD, pH 7.5, 25°C, 40 mM (NH4)2SO4, 0.5 mg/ml bovine serum albumin
691053
Lactococcus lactis
deoxyadenosine + thymine
-
-
-
r
2.4.2.6
cytosine + deoxyadenosine
27.7% conversion after 4 h at standard reaction conditions: 2 mM each substrate, 0.013 U NTD, pH 7.5, 25°C, 40 mM (NH4)2SO4, 0.5 mg/ml bovine serum albumin
691053
Lactococcus lactis
deoxycytidine + adenine
-
-
-
r
2.4.2.6
cytosine + deoxyguanosine
25.7% conversion after 4 h at standard reaction conditions: 2 mM each substrate, 0.013 U NTD, pH 7.5, 25°C, 40 mM (NH4)2SO4, 0.5 mg/ml bovine serum albumin
691053
Lactococcus lactis
deoxycytidine + guanine
-
-
-
r
2.4.2.6
cytosine + deoxyinosine
32.0% conversion after 4 h at standard reaction conditions: 2 mM each substrate, 0.013 U NTD, pH 7.5, 25°C, 40 mM (NH4)2SO4, 0.5 mg/ml bovine serum albumin
691053
Lactococcus lactis
deoxycytidine + hypoxanthine
-
-
-
r
2.4.2.6
cytosine + deoxyuridine
62.9% conversion after 4 h at standard reaction conditions: 2 mM each substrate, 0.013 U NTD, pH 7.5, 25°C, 40 mM (NH4)2SO4, 0.5 mg/ml bovine serum albumin
691053
Lactococcus lactis
deoxycytidine + uracil
-
-
-
r
2.4.2.6
cytosine + thymidine
53.5% conversion after 4 h at standard reaction conditions: 2 mM each substrate, 0.013 U NTD, pH 7.5, 25°C, 40 mM (NH4)2SO4, 0.5 mg/ml bovine serum albumin
691053
Lactococcus lactis
deoxycytidine + thymine
-
-
-
r
2.4.2.6
cytosine + thymidine
standard reaction for cofactor analyses as well as activating compound and inhibitor studies, in presence of 40 mM ammonium sulfate and 0.5 mg/ml bovine serum albumin
691053
Lactococcus lactis
deoxycytidine + thymine
-
-
-
r
2.4.2.6
hypoxanthine + deoxyadenosine
38.0% conversion after 4 h at standard reaction conditions: 2 mM substrate, 0.013 U NTD, pH 7.5, 25°C, 40 mM (NH4)2SO4, 0.5 mg/ml bovine serum albumin
691053
Lactococcus lactis
deoxyinosine + adenine
-
-
-
r
2.4.2.6
hypoxanthine + deoxycytidine
51.9% conversion after 4 h at standard reaction conditions: 2 mM each substrate, 0.013 U NTD, pH 7.5, 25°C, 40 mM (NH4)2SO4, 0.5 mg/ml bovine serum albumin
691053
Lactococcus lactis
deoxyinosine + cytosine
-
-
-
r
2.4.2.6
hypoxanthine + deoxyguanosine
25.2% conversion after 4 h at standard reaction conditions: 2 mM substrate, 0.013 U NTD, pH 7.5, 25°C, 40 mM (NH4)2SO4, 0.5 mg/ml bovine serum albumin
691053
Lactococcus lactis
deoxyinosine + guanine
-
-
-
r
2.4.2.6
hypoxanthine + deoxyuridine
65.6% conversion after 4 h at standard reaction conditions: 2 mM substrate, 0.013 U NTD, pH 7.5, 25°C, 40 mM (NH4)2SO4, 0.5 mg/ml bovine serum albumin
691053
Lactococcus lactis
deoxyinosine + uracil
-
-
-
r
2.4.2.6
hypoxanthine + thymidine
76.8% conversion after 4 h at standard reaction conditions: 2 mM substrate, 0.013 U NTD, pH 7.5, 25°C, 40 mM (NH4)2SO4, 0.5 mg/ml bovine serum albumin
691053
Lactococcus lactis
deoxyinosine + thymine
-
-
-
r
2.4.2.6
additional information
preferably purine 2-deoxyribonucleoside synthesis compared to pyrimidine deoxyribonucleoside synthesis although both are catalysed
691053
Lactococcus lactis
?
-
-
-
-
2.4.2.6
additional information
ribonucleoside, arabinosylnucleoside and 2,3-dideoxyribonucleoside are no substrates for base-exchange by YejD
691053
Lactococcus lactis
?
-
-
-
-
2.4.2.6
purine base + 2-deoxy-D-ribosyl-base
-
691053
Lactococcus lactis
purine deoxyribonucleoside + base
preferred over pyrimidine deoxyribonucleoside synthesis
-
-
r
2.4.2.6
pyrimidine base + 2-deoxy-D-ribosyl-base
-
691053
Lactococcus lactis
pyrimidine deoxyribonucleoside + base
-
-
-
r
2.4.2.6
thymine + deoxyadenosine
22.7% conversion after 4 h at standard reaction conditions: 2 mM each substrate, 0.013 U NTD, pH 7.5, 25°C, 40 mM (NH4)2SO4, 0.5 mg/ml bovine serum albumin
691053
Lactococcus lactis
thymidine + adenine
-
-
-
r
2.4.2.6
thymine + deoxycytidine
38.3% conversion after 4 h at standard reaction conditions: 2 mM each substrate, 0.013 U NTD, pH 7.5, 25°C, 40 mM (NH4)2SO4, 0.5 mg/ml bovine serum albumin
691053
Lactococcus lactis
thymidine + cytosine
-
-
-
r
2.4.2.6
thymine + deoxyguanosine
17.7% conversion after 4 h at standard reaction conditions: 2 mM each substrate, 0.013 U NTD, pH 7.5, 25°C, 40 mM (NH4)2SO4, 0.5 mg/ml bovine serum albumin
691053
Lactococcus lactis
thymidine + guanine
-
-
-
r
2.4.2.6
thymine + deoxyinosine
19.5% conversion after 4 h at standard reaction conditions: 2 mM each substrate, 0.013 U NTD, pH 7.5, 25°C, 40 mM (NH4)2SO4, 0.5 mg/ml bovine serum albumin, equilibrium reached after 2 h
691053
Lactococcus lactis
thymidine + hypoxanthine
-
-
-
r
2.4.2.6
thymine + deoxyuridine
53.7% conversion after 4 h at standard reaction conditions: 2 mM each substrate, 0.013 U NTD, pH 7.5, 25°C, 40 mM (NH4)2SO4, 0.5 mg/ml bovine serum albumin
691053
Lactococcus lactis
thymidine + uracil
-
-
-
r
2.4.2.6
uracil + deoxyadenosine
17.1% conversion after 4 h at standard reaction conditions: 2 mM each substrate, 0.013 U NTD, pH 7.5, 25°C, 40 mM (NH4)2SO4, 0.5 mg/ml bovine serum albumin
691053
Lactococcus lactis
deoxyuridine + adenine
-
-
-
r
2.4.2.6
uracil + deoxycytidine
37.7% conversion after 4 h at standard reaction conditions: 2 mM each substrate, 0.013 U NTD, pH 7.5, 25°C, 40 mM (NH4)2SO4, 0.5 mg/ml bovine serum albumin
691053
Lactococcus lactis
deoxyuridine + cytosine
-
-
-
r
2.4.2.6
uracil + deoxyguanosine
16.1% conversion after 4 h at standard reaction conditions: 2 mM each substrate, 0.013 U NTD, pH 7.5, 25°C, 40 mM (NH4)2SO4, 0.5 mg/ml bovine serum albumin
691053
Lactococcus lactis
deoxyuridine + guanine
-
-
-
r
2.4.2.6
uracil + deoxyinosine
23.8% conversion after 4 h at standard reaction conditions: 2 mM each substrate, 0.013 U NTD, pH 7.5, 25°C, 40 mM (NH4)2SO4, 0.5 mg/ml bovine serum albumin
691053
Lactococcus lactis
deoxyuridine + hypoxanthine
-
-
-
r
2.4.2.6
uracil + thymidine
43.0% conversion after 4 h at standard reaction conditions: 2 mM each substrate, 0.013 U NTD, pH 7.5, 25°C, 40 mM (NH4)2SO4, 0.5 mg/ml bovine serum albumin
691053
Lactococcus lactis
deoxyuridine + thymine
-
-
-
r
Subunits
EC Number
Subunits
Commentary
Organism
2.4.2.6
homotetramer
4 * 17250, elution as single peak from the preparative gel filtration column
Lactococcus lactis
Temperature Optimum [°C]
EC Number
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
2.4.2.6
30
-
assay at, pH 7.5
Lactococcus lactis
Temperature Stability [°C]
EC Number
Temperature Stability Minimum [°C]
Temperature Stability Maximum [°C]
Commentary
Organism
2.4.2.6
additional information
-
(NH4)2SO4 is more thermostabilizing than KCl
Lactococcus lactis
2.4.2.6
25
-
pH 7.5, 20 min, 60% activity lost, in presence of 10 mM (NH4)2SO4 only 20% activity lost, in presence of 500 mM KCL only 6% activity lost
Lactococcus lactis
2.4.2.6
30
-
pH 7.5, 20 min, in presence of 500 mM (NH4)2SO4 fully active
Lactococcus lactis
2.4.2.6
35
-
and more, pH 7.5, 20 min, 75% activity lost
Lactococcus lactis
pH Optimum
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
2.4.2.6
7.5
-
assay at
Lactococcus lactis
Activating Compound (protein specific)
EC Number
Activating Compound
Commentary
Organism
Structure
2.4.2.6
K+
K+-activated type II enzyme: binding of K+ is required to enhance activity through induced conformational change
Lactococcus lactis
2.4.2.6
additional information
does not require phosphate derivatives such as ATP, thiamine diphosphate, pyridoxal phosphate to complete 2-deoxyribose transfer
Lactococcus lactis
Cloned(Commentary) (protein specific)
EC Number
Commentary
Organism
2.4.2.6
in pET-22b(+) for expression in Escherichia coli BL21(DE3); in pGEM-T for cloning
Lactococcus lactis
Inhibitors (protein specific)
EC Number
Inhibitors
Commentary
Organism
Structure
2.4.2.6
Ca2+
inhibits base exchange activity at 1 mM
Lactococcus lactis
2.4.2.6
CO2
reduces base exchange activity by 87-99% at 1 mM
Lactococcus lactis
2.4.2.6
Cu2+
reduces base exchange activity by 87-99% at 1 mM
Lactococcus lactis
2.4.2.6
Mg2+
inhibits base exchange activity at 1mM
Lactococcus lactis
2.4.2.6
Mn2+
reduces base exchange activity by 87-99% at 1 mM
Lactococcus lactis
2.4.2.6
Zn2+
reduces base exchange activity by 87-99% at 1 mM
Lactococcus lactis
Metals/Ions (protein specific)
EC Number
Metals/Ions
Commentary
Organism
Structure
2.4.2.6
K+
enhances base exchange activity and thermostability, 1.66-fold activation by 1 mM KCl (reaches plateau at 20 mM)
Lactococcus lactis
2.4.2.6
additional information
enzymatic activity depends on monovalent cations as cofactors rather than catalysts; K+ and NH4+ have no synergistic effects; Li+ and Na+ do not activate
Lactococcus lactis
2.4.2.6
NH4+
enhances base exchange activity and thermostability, 1.43-fold activation by 1 mM (NH4)2SO4 (reaches plateau at 20 mM)
Lactococcus lactis
2.4.2.6
Rb+
enhances base exchange activity, 1.51-fold activation by 1 mM RbCl
Lactococcus lactis
Molecular Weight [Da] (protein specific)
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
2.4.2.6
17250
-
4 * 17250, elution as single peak from the preparative gel filtration column
Lactococcus lactis
2.4.2.6
18290
-
theoretical molecular weight
Lactococcus lactis
2.4.2.6
25500
-
according to migration in SDS-12.5% polyacrylamide gel electrophoresis
Lactococcus lactis
2.4.2.6
69000
-
gel filtration chromatography
Lactococcus lactis
Natural Substrates/ Products (Substrates) (protein specific)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
2.4.2.6
additional information
Lactococcus lactis
preferably purine 2-deoxyribonucleoside synthesis compared to pyrimidine deoxyribonucleoside synthesis although both are catalysed
?
-
-
-
2.4.2.6
purine base + 2-deoxy-D-ribosyl-base
Lactococcus lactis
-
purine deoxyribonucleoside + base
preferred over pyrimidine deoxyribonucleoside synthesis
-
r
2.4.2.6
pyrimidine base + 2-deoxy-D-ribosyl-base
Lactococcus lactis
-
pyrimidine deoxyribonucleoside + base
-
-
r
Purification (Commentary) (protein specific)
EC Number
Commentary
Organism
2.4.2.6
preclearing of bacterial lysate with 40% ammonium sulfate and subsequent precipitation of proteins with 90% ammonium sulfate, gel filtration of dissolved protein on Sephacryl S-200 column, combination of fractions with N-deoxyribosyltransferase activity, precipitation of proteins with 90% ammonium sulfate, dialysis of dissolved proteins and concentration by ultrafiltration, second gel filtration on Sephacryl S-200 column, combination of active fractions and concentration by ultrafiltration, recovery of 70.4% of activity, storage at -20°C in 50% glycerol
Lactococcus lactis
Specific Activity [micromol/min/mg] (protein specific)
EC Number
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
2.4.2.6
additional information
-
0.9% in presence of 1 mM ZnCl2; 102% in presence of 1 mM NaCl; 105% in presence of 1 mM CsCl; 106% in presence of 1 mM NaSO4; 12.9% in presence of 1 mM CoCl2; 143% in presence of 1 mM NH4Cl; 151% in presence of 1 mM RbCl; 163% in presence of 1 mM (NH4)2SO4; 165% in presence of 1 mM K2SO4; 166% in presence of 1 mM KCl; 166% in presence of 1 mM (NH4)2SO4 and 1 mM KCl; 192% in presence of 10 mM KCl; 194% in presence of 10 mM (NH4)2SO4; 3.2% in presence of 1 mM CuSO4; 51.8% in presence of 1 mM CaCl2; 66.3% in presence of 1 mM MgCl2; 66% in presence of 1 mM MgSO4; 68.3% in presence of 1 mM FeCl2; 97.7% in presence of 1 mM LiCl
Lactococcus lactis
2.4.2.6
6.67
-
equals 100%, purified protein, in presence of 0.5 mg/ml bovine serum albumin, pH 7.5, 10 min, 25°C
Lactococcus lactis
Substrates and Products (Substrate) (protein specific)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
2.4.2.6
adenine + deoxycytidine
64.0% conversion after 4 h at standard reaction conditions: 2 mM each substrate, 0.013 U NTD, pH 7.5, 25°C, 40 mM (NH4)2SO4, 0.5 mg/ml bovine serum albumin
691053
Lactococcus lactis
deoxyadenosine + cytosine
-
-
-
r
2.4.2.6
adenine + deoxyguanosine
46.6% conversion after 4 h at standard reaction conditions: 2 mM each substrate, 0.013 U NTD, pH 7.5, 25°C, 40 mM (NH4)2SO4, 0.5 mg/ml bovine serum albumin
691053
Lactococcus lactis
deoxyadenosine + guanine
-
-
-
r
2.4.2.6
adenine + deoxyinosine
57.3% conversion after 4 h at standard reaction conditions: 2 mM each substrate, 0.013 U NTD, pH 7.5, 25°C, 40 mM (NH4)2SO4, 0.5 mg/ml bovine serum albumin
691053
Lactococcus lactis
deoxyadenosine + hypoxanthine
-
-
-
r
2.4.2.6
adenine + deoxyuridine
75.4% conversion after 4 h at standard reaction conditions: 2 mM each substrate, 0.013 U NTD, pH 7.5, 25°C, 40 mM (NH4)2SO4, 0.5 mg/ml bovine serum albumin
691053
Lactococcus lactis
deoxyadenosine + uracil
-
-
-
r
2.4.2.6
adenine + thymidine
74.6% conversion after 4 h at standard reaction conditions: 2 mM each substrate, 0.013 U NTD, pH 7.5, 25°C, 40 mM (NH4)2SO4, 0.5 mg/ml bovine serum albumin
691053
Lactococcus lactis
deoxyadenosine + thymine
-
-
-
r
2.4.2.6
cytosine + deoxyadenosine
27.7% conversion after 4 h at standard reaction conditions: 2 mM each substrate, 0.013 U NTD, pH 7.5, 25°C, 40 mM (NH4)2SO4, 0.5 mg/ml bovine serum albumin
691053
Lactococcus lactis
deoxycytidine + adenine
-
-
-
r
2.4.2.6
cytosine + deoxyguanosine
25.7% conversion after 4 h at standard reaction conditions: 2 mM each substrate, 0.013 U NTD, pH 7.5, 25°C, 40 mM (NH4)2SO4, 0.5 mg/ml bovine serum albumin
691053
Lactococcus lactis
deoxycytidine + guanine
-
-
-
r
2.4.2.6
cytosine + deoxyinosine
32.0% conversion after 4 h at standard reaction conditions: 2 mM each substrate, 0.013 U NTD, pH 7.5, 25°C, 40 mM (NH4)2SO4, 0.5 mg/ml bovine serum albumin
691053
Lactococcus lactis
deoxycytidine + hypoxanthine
-
-
-
r
2.4.2.6
cytosine + deoxyuridine
62.9% conversion after 4 h at standard reaction conditions: 2 mM each substrate, 0.013 U NTD, pH 7.5, 25°C, 40 mM (NH4)2SO4, 0.5 mg/ml bovine serum albumin
691053
Lactococcus lactis
deoxycytidine + uracil
-
-
-
r
2.4.2.6
cytosine + thymidine
53.5% conversion after 4 h at standard reaction conditions: 2 mM each substrate, 0.013 U NTD, pH 7.5, 25°C, 40 mM (NH4)2SO4, 0.5 mg/ml bovine serum albumin
691053
Lactococcus lactis
deoxycytidine + thymine
-
-
-
r
2.4.2.6
cytosine + thymidine
standard reaction for cofactor analyses as well as activating compound and inhibitor studies, in presence of 40 mM ammonium sulfate and 0.5 mg/ml bovine serum albumin
691053
Lactococcus lactis
deoxycytidine + thymine
-
-
-
r
2.4.2.6
hypoxanthine + deoxyadenosine
38.0% conversion after 4 h at standard reaction conditions: 2 mM substrate, 0.013 U NTD, pH 7.5, 25°C, 40 mM (NH4)2SO4, 0.5 mg/ml bovine serum albumin
691053
Lactococcus lactis
deoxyinosine + adenine
-
-
-
r
2.4.2.6
hypoxanthine + deoxycytidine
51.9% conversion after 4 h at standard reaction conditions: 2 mM each substrate, 0.013 U NTD, pH 7.5, 25°C, 40 mM (NH4)2SO4, 0.5 mg/ml bovine serum albumin
691053
Lactococcus lactis
deoxyinosine + cytosine
-
-
-
r
2.4.2.6
hypoxanthine + deoxyguanosine
25.2% conversion after 4 h at standard reaction conditions: 2 mM substrate, 0.013 U NTD, pH 7.5, 25°C, 40 mM (NH4)2SO4, 0.5 mg/ml bovine serum albumin
691053
Lactococcus lactis
deoxyinosine + guanine
-
-
-
r
2.4.2.6
hypoxanthine + deoxyuridine
65.6% conversion after 4 h at standard reaction conditions: 2 mM substrate, 0.013 U NTD, pH 7.5, 25°C, 40 mM (NH4)2SO4, 0.5 mg/ml bovine serum albumin
691053
Lactococcus lactis
deoxyinosine + uracil
-
-
-
r
2.4.2.6
hypoxanthine + thymidine
76.8% conversion after 4 h at standard reaction conditions: 2 mM substrate, 0.013 U NTD, pH 7.5, 25°C, 40 mM (NH4)2SO4, 0.5 mg/ml bovine serum albumin
691053
Lactococcus lactis
deoxyinosine + thymine
-
-
-
r
2.4.2.6
additional information
preferably purine 2-deoxyribonucleoside synthesis compared to pyrimidine deoxyribonucleoside synthesis although both are catalysed
691053
Lactococcus lactis
?
-
-
-
-
2.4.2.6
additional information
ribonucleoside, arabinosylnucleoside and 2,3-dideoxyribonucleoside are no substrates for base-exchange by YejD
691053
Lactococcus lactis
?
-
-
-
-
2.4.2.6
purine base + 2-deoxy-D-ribosyl-base
-
691053
Lactococcus lactis
purine deoxyribonucleoside + base
preferred over pyrimidine deoxyribonucleoside synthesis
-
-
r
2.4.2.6
pyrimidine base + 2-deoxy-D-ribosyl-base
-
691053
Lactococcus lactis
pyrimidine deoxyribonucleoside + base
-
-
-
r
2.4.2.6
thymine + deoxyadenosine
22.7% conversion after 4 h at standard reaction conditions: 2 mM each substrate, 0.013 U NTD, pH 7.5, 25°C, 40 mM (NH4)2SO4, 0.5 mg/ml bovine serum albumin
691053
Lactococcus lactis
thymidine + adenine
-
-
-
r
2.4.2.6
thymine + deoxycytidine
38.3% conversion after 4 h at standard reaction conditions: 2 mM each substrate, 0.013 U NTD, pH 7.5, 25°C, 40 mM (NH4)2SO4, 0.5 mg/ml bovine serum albumin
691053
Lactococcus lactis
thymidine + cytosine
-
-
-
r
2.4.2.6
thymine + deoxyguanosine
17.7% conversion after 4 h at standard reaction conditions: 2 mM each substrate, 0.013 U NTD, pH 7.5, 25°C, 40 mM (NH4)2SO4, 0.5 mg/ml bovine serum albumin
691053
Lactococcus lactis
thymidine + guanine
-
-
-
r
2.4.2.6
thymine + deoxyinosine
19.5% conversion after 4 h at standard reaction conditions: 2 mM each substrate, 0.013 U NTD, pH 7.5, 25°C, 40 mM (NH4)2SO4, 0.5 mg/ml bovine serum albumin, equilibrium reached after 2 h
691053
Lactococcus lactis
thymidine + hypoxanthine
-
-
-
r
2.4.2.6
thymine + deoxyuridine
53.7% conversion after 4 h at standard reaction conditions: 2 mM each substrate, 0.013 U NTD, pH 7.5, 25°C, 40 mM (NH4)2SO4, 0.5 mg/ml bovine serum albumin
691053
Lactococcus lactis
thymidine + uracil
-
-
-
r
2.4.2.6
uracil + deoxyadenosine
17.1% conversion after 4 h at standard reaction conditions: 2 mM each substrate, 0.013 U NTD, pH 7.5, 25°C, 40 mM (NH4)2SO4, 0.5 mg/ml bovine serum albumin
691053
Lactococcus lactis
deoxyuridine + adenine
-
-
-
r
2.4.2.6
uracil + deoxycytidine
37.7% conversion after 4 h at standard reaction conditions: 2 mM each substrate, 0.013 U NTD, pH 7.5, 25°C, 40 mM (NH4)2SO4, 0.5 mg/ml bovine serum albumin
691053
Lactococcus lactis
deoxyuridine + cytosine
-
-
-
r
2.4.2.6
uracil + deoxyguanosine
16.1% conversion after 4 h at standard reaction conditions: 2 mM each substrate, 0.013 U NTD, pH 7.5, 25°C, 40 mM (NH4)2SO4, 0.5 mg/ml bovine serum albumin
691053
Lactococcus lactis
deoxyuridine + guanine
-
-
-
r
2.4.2.6
uracil + deoxyinosine
23.8% conversion after 4 h at standard reaction conditions: 2 mM each substrate, 0.013 U NTD, pH 7.5, 25°C, 40 mM (NH4)2SO4, 0.5 mg/ml bovine serum albumin
691053
Lactococcus lactis
deoxyuridine + hypoxanthine
-
-
-
r
2.4.2.6
uracil + thymidine
43.0% conversion after 4 h at standard reaction conditions: 2 mM each substrate, 0.013 U NTD, pH 7.5, 25°C, 40 mM (NH4)2SO4, 0.5 mg/ml bovine serum albumin
691053
Lactococcus lactis
deoxyuridine + thymine
-
-
-
r
Subunits (protein specific)
EC Number
Subunits
Commentary
Organism
2.4.2.6
homotetramer
4 * 17250, elution as single peak from the preparative gel filtration column
Lactococcus lactis
Temperature Optimum [°C] (protein specific)
EC Number
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
2.4.2.6
30
-
assay at, pH 7.5
Lactococcus lactis
Temperature Stability [°C] (protein specific)
EC Number
Temperature Stability Minimum [°C]
Temperature Stability Maximum [°C]
Commentary
Organism
2.4.2.6
additional information
-
(NH4)2SO4 is more thermostabilizing than KCl
Lactococcus lactis
2.4.2.6
25
-
pH 7.5, 20 min, 60% activity lost, in presence of 10 mM (NH4)2SO4 only 20% activity lost, in presence of 500 mM KCL only 6% activity lost
Lactococcus lactis
2.4.2.6
30
-
pH 7.5, 20 min, in presence of 500 mM (NH4)2SO4 fully active
Lactococcus lactis
2.4.2.6
35
-
and more, pH 7.5, 20 min, 75% activity lost
Lactococcus lactis
pH Optimum (protein specific)
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
2.4.2.6
7.5
-
assay at
Lactococcus lactis