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Literature summary extracted from

  • Lim, J.H.; Choi, J.; Han, S.J.; Kim, S.H.; Hwang, H.Z.; Jin, D.K.; Ahn, B.Y.; Han, Y.S.
    Molecular cloning and characterization of thermostable DNA ligase from Aquifex pyrophilus, a hyperthermophilic bacterium (2001), Extremophiles, 5, 161-168.
    View publication on PubMed

Activating Compound

EC Number Activating Compound Comment Organism Structure
6.5.1.2 KCl highest activity at 150 mM Aquifex pyrophilus
6.5.1.2 NaCl highest activity at 200 mM Aquifex pyrophilus
6.5.1.2 NH4Cl highest activity at 200 mM Aquifex pyrophilus
6.5.1.2 potassium glutamate highest activity at 350 mM Aquifex pyrophilus

Cloned(Commentary)

EC Number Cloned (Comment) Organism
6.5.1.2 expressed in Escherichia coli Aquifex pyrophilus

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
6.5.1.2 Ca2+ 40% of activity obtained with Mg2+ Aquifex pyrophilus
6.5.1.2 Cu2+ 30% of activity obtained with Mg2+ Aquifex pyrophilus
6.5.1.2 K+ highest activity below 10 mM Thermus aquaticus
6.5.1.2 Mg2+ required Aquifex pyrophilus
6.5.1.2 Mn2+ 50% of activity obtained with Mg2+ Aquifex pyrophilus
6.5.1.2 Na+ inhibitory Thermus aquaticus
6.5.1.2 NH4+ highest activity below 10 mM Thermus aquaticus
6.5.1.2 Ni2+ 10% of activity obtained with Mg2+ Aquifex pyrophilus
6.5.1.2 Zn2+ 5% of activity obtained with Mg2+ Aquifex pyrophilus

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
6.5.1.2 82000
-
SDS-PAGE Aquifex pyrophilus

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
6.5.1.2 NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m Thermus aquaticus DNA-repair, phosphodiester bond formation between adjacent 5'-phosphate and 3'-hydroxyl groups in double-stranded DNA AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
-
?
6.5.1.2 NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m Escherichia coli DNA-repair, phosphodiester bond formation between adjacent 5'-phosphate and 3'-hydroxyl groups in double-stranded DNA AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
-
?
6.5.1.2 NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m Aquifex pyrophilus DNA-repair, phosphodiester bond formation between adjacent 5'-phosphate and 3'-hydroxyl groups in double-stranded DNA AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
-
?

Organism

EC Number Organism UniProt Comment Textmining
6.5.1.2 Aquifex pyrophilus Q9F150
-
-
6.5.1.2 Escherichia coli
-
-
-
6.5.1.2 Thermus aquaticus
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
6.5.1.2 recombinant protein Aquifex pyrophilus

Reaction

EC Number Reaction Comment Organism Reaction ID
6.5.1.2 ATP + (deoxyribonucleotide)n-3'-hydroxyl + 5'-phospho-(deoxyribonucleotide)m = (deoxyribonucleotide)n+m + AMP + beta-nicotinamide D-nucleotide 3 steps of reaction: 1. adenylation of the ligase in the presence of NAD+, 2. transferring the adenylate moiety to the 5'-phosphate of the nicked DNA substrate, 3. sealing the nick through the formation of a phosphodiester bond Thermus aquaticus
6.5.1.2 ATP + (deoxyribonucleotide)n-3'-hydroxyl + 5'-phospho-(deoxyribonucleotide)m = (deoxyribonucleotide)n+m + AMP + beta-nicotinamide D-nucleotide 3 steps of reaction: 1. adenylation of the ligase in the presence of NAD+, 2. transferring the adenylate moiety to the 5'-phosphate of the nicked DNA substrate, 3. sealing the nick through the formation of a phosphodiester bond Escherichia coli
6.5.1.2 ATP + (deoxyribonucleotide)n-3'-hydroxyl + 5'-phospho-(deoxyribonucleotide)m = (deoxyribonucleotide)n+m + AMP + beta-nicotinamide D-nucleotide 3 steps of reaction: 1. adenylation of the ligase in the presence of NAD+, 2. transferring the adenylate moiety to the 5'-phosphate of the nicked DNA substrate, 3. sealing the nick through the formation of a phosphodiester bond Aquifex pyrophilus

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
6.5.1.2 NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
-
Thermus aquaticus AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
-
?
6.5.1.2 NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
-
Escherichia coli AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
-
?
6.5.1.2 NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
-
Aquifex pyrophilus AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
-
?
6.5.1.2 NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m DNA-repair, phosphodiester bond formation between adjacent 5'-phosphate and 3'-hydroxyl groups in double-stranded DNA Thermus aquaticus AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
-
?
6.5.1.2 NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m DNA-repair, phosphodiester bond formation between adjacent 5'-phosphate and 3'-hydroxyl groups in double-stranded DNA Escherichia coli AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
-
?
6.5.1.2 NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m DNA-repair, phosphodiester bond formation between adjacent 5'-phosphate and 3'-hydroxyl groups in double-stranded DNA Aquifex pyrophilus AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
-
?

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
6.5.1.2 25
-
-
Escherichia coli
6.5.1.2 50
-
-
Thermus aquaticus
6.5.1.2 65
-
above 65°C Aquifex pyrophilus

Temperature Range [°C]

EC Number Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
6.5.1.2 5 50
-
Escherichia coli
6.5.1.2 20 90
-
Thermus aquaticus
6.5.1.2 30 90
-
Aquifex pyrophilus

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
6.5.1.2 95
-
50% of activity lost within 15 min Thermus aquaticus
6.5.1.2 95
-
50% of activity lost within 5 min Escherichia coli
6.5.1.2 95
-
60 min, less than 25% of activity lost Aquifex pyrophilus

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
6.5.1.2 8 8.6
-
Aquifex pyrophilus

Cofactor

EC Number Cofactor Comment Organism Structure
6.5.1.2 NAD+ required Thermus aquaticus
6.5.1.2 NAD+ required Escherichia coli
6.5.1.2 NAD+ required Aquifex pyrophilus