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Literature summary for 6.5.1.2 extracted from

  • Takahashi, M.; Yamaguchi, E.; Uchida, T.
    Thermophilic DNA ligase. Purification and properties of the enzyme from Thermus thermophilus HB8 (1984), J. Biol. Chem., 259, 10041-10047.
    View publication on PubMed

Inhibitors

Inhibitors Comment Organism Structure
Na+ above 20-150 mM Thermus thermophilus

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.0000185
-
NAD+
-
Thermus thermophilus
0.000025
-
5'-phosphoryl termini of poly(dA-dT)
-
Thermus thermophilus
0.14
-
(dT)10
-
Thermus thermophilus

Metals/Ions

Metals/Ions Comment Organism Structure
K+ activates at 1-150 mM Thermus thermophilus
Mg2+ requires divalent cations, Mn2+ or Mg2+ Thermus thermophilus
Mg2+ optimal concentration: 5-9 mM Thermus thermophilus
Mn2+ requires divalent cations, Mn2+ or Mg2+ Thermus thermophilus
Mn2+ optimal concentration: 3-6 mM Thermus thermophilus
NH4+ activates at 1-150 mM Thermus thermophilus

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
78500
-
1 * 78500, SDS-PAGE Thermus thermophilus
80400
-
sedimentation equilibrium ultracentrifugation Thermus thermophilus

Organism

Organism UniProt Comment Textmining
Thermus thermophilus
-
-
-

Purification (Commentary)

Purification (Comment) Organism
-
Thermus thermophilus

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
additional information
-
-
Thermus thermophilus

Storage Stability

Storage Stability Organism
4°C, stable for several months Thermus thermophilus

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
ATP + (dT)10
-
Thermus thermophilus ?
-
?
ATP + 5'-phosphoryl termini of poly(dA-dT)
-
Thermus thermophilus ?
-
?
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m (dT)8, (dT)10, (dT)12-18 Thermus thermophilus AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
-
?
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m cohesive-end DNA restriction fragments Thermus thermophilus AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
-
?
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m nick-closing activity Thermus thermophilus AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
-
?

Subunits

Subunits Comment Organism
monomer 1 * 78500, SDS-PAGE Thermus thermophilus

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
24 37 cohesive-end DNA restriction fragments Thermus thermophilus
27.7
-
p(dT)8, joining of thymidylate oligomers in the presence of poly(dA) as a template Thermus thermophilus
34.5
-
p(dT)10, joining of thymidylate oligomers in the presence of poly(dA) as a template Thermus thermophilus
37
-
p(dT)12-18, joining of thymidylate oligomers in the presence of poly(dA) as a template Thermus thermophilus
65 72 nick-closing activity Thermus thermophilus

Temperature Range [°C]

Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
15 85 nick-closing activity observed in the range Thermus thermophilus

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
4
-
activity decreases remarkably below 4°C Thermus thermophilus
24
-
stable for 1 week Thermus thermophilus
37
-
stable for 1 week Thermus thermophilus
65
-
2 days, 50% loss of activity Thermus thermophilus

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.4 7.6 Tris-HCl buffer Thermus thermophilus
8
-
collidine/HCl buffer Thermus thermophilus