Any feedback?
Please rate this page
(literature.php)
(0/150)

BRENDA support

Literature summary for 2.4.2.6 extracted from

  • Miyamoto, Y.; Masaki, T.; Chohnan, S.
    Characterization of N-deoxyribosyltransferase from Lactococcus lactis subsp. lactis (2007), Biochim. Biophys. Acta, 1774, 1323-1330.
    View publication on PubMed
Search for more literature for 2.4.2.6

Activating Compound

Activating Compound Comment Organism Structure
K+ K+-activated type II enzyme: binding of K+ is required to enhance activity through induced conformational change Lactococcus lactis
additional information does not require phosphate derivatives such as ATP, thiamine diphosphate, pyridoxal phosphate to complete 2-deoxyribose transfer Lactococcus lactis

Cloned(Commentary)

Cloned (Comment) Organism
in pET-22b(+) for expression in Escherichia coli BL21(DE3) Lactococcus lactis
in pGEM-T for cloning Lactococcus lactis

Inhibitors

Inhibitors Comment Organism Structure
Ca2+ inhibits base exchange activity at 1 mM Lactococcus lactis
CO2 reduces base exchange activity by 87-99% at 1 mM Lactococcus lactis
Cu2+ reduces base exchange activity by 87-99% at 1 mM Lactococcus lactis
Mg2+ inhibits base exchange activity at 1mM Lactococcus lactis
Mn2+ reduces base exchange activity by 87-99% at 1 mM Lactococcus lactis
Zn2+ reduces base exchange activity by 87-99% at 1 mM Lactococcus lactis

Metals/Ions

Metals/Ions Comment Organism Structure
K+ enhances base exchange activity and thermostability, 1.66-fold activation by 1 mM KCl (reaches plateau at 20 mM) Lactococcus lactis
additional information enzymatic activity depends on monovalent cations as cofactors rather than catalysts Lactococcus lactis
additional information K+ and NH4+ have no synergistic effects Lactococcus lactis
additional information Li+ and Na+ do not activate Lactococcus lactis
NH4+ enhances base exchange activity and thermostability, 1.43-fold activation by 1 mM (NH4)2SO4 (reaches plateau at 20 mM) Lactococcus lactis
Rb+ enhances base exchange activity, 1.51-fold activation by 1 mM RbCl Lactococcus lactis

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
17250
-
 4 * 17250, elution as single peak from the preparative gel filtration column Lactococcus lactis
18290
-
 theoretical molecular weight Lactococcus lactis
25500
-
 according to migration in SDS-12.5% polyacrylamide gel electrophoresis Lactococcus lactis
69000
-
 gel filtration chromatography Lactococcus lactis

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
additional information Lactococcus lactis preferably purine 2-deoxyribonucleoside synthesis compared to pyrimidine deoxyribonucleoside synthesis although both are catalysed ?
-
 ?
purine base + 2-deoxy-D-ribosyl-base Lactococcus lactis
-
 purine deoxyribonucleoside + base preferred over pyrimidine deoxyribonucleoside synthesis r
pyrimidine base + 2-deoxy-D-ribosyl-base Lactococcus lactis
-
 pyrimidine deoxyribonucleoside + base
-
 r

Organism

Organism UniProt Comment Textmining
Lactococcus lactis
-
 subsp. lactis JCM5808
-

Purification (Commentary)

Purification (Comment) Organism
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]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
additional information
-
 0.9% in presence of 1 mM ZnCl2 Lactococcus lactis
additional information
-
 102% in presence of 1 mM NaCl Lactococcus lactis
additional information
-
 105% in presence of 1 mM CsCl Lactococcus lactis
additional information
-
 106% in presence of 1 mM NaSO4 Lactococcus lactis
additional information
-
 12.9% in presence of 1 mM CoCl2 Lactococcus lactis
additional information
-
 143% in presence of 1 mM NH4Cl Lactococcus lactis
additional information
-
 151% in presence of 1 mM RbCl Lactococcus lactis
additional information
-
 163% in presence of 1 mM (NH4)2SO4 Lactococcus lactis
additional information
-
 165% in presence of 1 mM K2SO4 Lactococcus lactis
additional information
-
 166% in presence of 1 mM (NH4)2SO4 and 1 mM KCl Lactococcus lactis
additional information
-
 166% in presence of 1 mM KCl Lactococcus lactis
additional information
-
 192% in presence of 10 mM KCl Lactococcus lactis
additional information
-
 194% in presence of 10 mM (NH4)2SO4 Lactococcus lactis
additional information
-
 3.2% in presence of 1 mM CuSO4 Lactococcus lactis
additional information
-
 51.8% in presence of 1 mM CaCl2 Lactococcus lactis
additional information
-
 66.3% in presence of 1 mM MgCl2 Lactococcus lactis
additional information
-
 66% in presence of 1 mM MgSO4 Lactococcus lactis
additional information
-
 68.3% in presence of 1 mM FeCl2 Lactococcus lactis
additional information
-
 97.7% in presence of 1 mM LiCl Lactococcus lactis
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)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
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 Lactococcus lactis deoxyadenosine + cytosine
-
 r
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 Lactococcus lactis deoxyadenosine + guanine
-
 r
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 Lactococcus lactis deoxyadenosine + hypoxanthine
-
 r
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 Lactococcus lactis deoxyadenosine + uracil
-
 r
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 Lactococcus lactis deoxyadenosine + thymine
-
 r
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 Lactococcus lactis deoxycytidine + adenine
-
 r
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 Lactococcus lactis deoxycytidine + guanine
-
 r
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 Lactococcus lactis deoxycytidine + hypoxanthine
-
 r
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 Lactococcus lactis deoxycytidine + uracil
-
 r
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 Lactococcus lactis deoxycytidine + thymine
-
 r
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 Lactococcus lactis deoxycytidine + thymine
-
 r
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 Lactococcus lactis deoxyinosine + adenine
-
 r
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 Lactococcus lactis deoxyinosine + cytosine
-
 r
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 Lactococcus lactis deoxyinosine + guanine
-
 r
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 Lactococcus lactis deoxyinosine + uracil
-
 r
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 Lactococcus lactis deoxyinosine + thymine
-
 r
additional information preferably purine 2-deoxyribonucleoside synthesis compared to pyrimidine deoxyribonucleoside synthesis although both are catalysed Lactococcus lactis ?
-
 ?
additional information ribonucleoside, arabinosylnucleoside and 2,3-dideoxyribonucleoside are no substrates for base-exchange by YejD Lactococcus lactis ?
-
 ?
purine base + 2-deoxy-D-ribosyl-base
-
 Lactococcus lactis purine deoxyribonucleoside + base preferred over pyrimidine deoxyribonucleoside synthesis r
pyrimidine base + 2-deoxy-D-ribosyl-base
-
 Lactococcus lactis pyrimidine deoxyribonucleoside + base
-
 r
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 Lactococcus lactis thymidine + adenine
-
 r
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 Lactococcus lactis thymidine + cytosine
-
 r
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 Lactococcus lactis thymidine + guanine
-
 r
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 Lactococcus lactis thymidine + hypoxanthine
-
 r
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 Lactococcus lactis thymidine + uracil
-
 r
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 Lactococcus lactis deoxyuridine + adenine
-
 r
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 Lactococcus lactis deoxyuridine + cytosine
-
 r
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 Lactococcus lactis deoxyuridine + guanine
-
 r
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 Lactococcus lactis deoxyuridine + hypoxanthine
-
 r
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 Lactococcus lactis deoxyuridine + thymine
-
 r

Subunits

Subunits Comment Organism
homotetramer 4 * 17250, elution as single peak from the preparative gel filtration column Lactococcus lactis

Synonyms

Synonyms Comment Organism
N-deoxyribosyltransferase
-
 Lactococcus lactis
NTD
-
 Lactococcus lactis
nucleoside N-deoxyribosyltransferase
-
 Lactococcus lactis
trans-N-deoxyribosylase
-
 Lactococcus lactis
YejD
-
 Lactococcus lactis

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
30
-
 assay at, pH 7.5 Lactococcus lactis

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
additional information
-
 (NH4)2SO4 is more thermostabilizing than KCl Lactococcus lactis
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
30
-
 pH 7.5, 20 min, in presence of 500 mM (NH4)2SO4 fully active Lactococcus lactis
35
-
 and more, pH 7.5, 20 min, 75% activity lost Lactococcus lactis

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.5
-
 assay at Lactococcus lactis