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Literature summary extracted from

  • Tong, J.; Barany, F.; Cao, W.
    Ligation reaction specificities of an NAD+-dependent DNA ligase from the hyperthermophile Aquifex aeolicus (2000), Nucleic Acids Res., 28, 1447-1454.
    View publication on PubMedView publication on EuropePMC

Activating Compound

EC Number Activating Compound Comment Organism Structure
6.5.1.2 KCl 150 mM Aquifex aeolicus

Cloned(Commentary)

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

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
6.5.1.2 0.00008
-
 nicked DNA pH 7.6, 65°C Aquifex aeolicus

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
6.5.1.2 Ca2+ supports formation of DNA-adenylate intermediate Aquifex aeolicus
6.5.1.2 Mg2+ 5 mM Aquifex aeolicus
6.5.1.2 Mn2+ 5 mM, most effective metal cofactor Aquifex aeolicus
6.5.1.2 Ni2+ supports formation of DNA-adenylate intermediate Aquifex aeolicus

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 Aquifex aeolicus 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 aeolicus O66880
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
6.5.1.2
-
 Aquifex aeolicus

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 Aquifex aeolicus
a

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
6.5.1.2 ATP + nicked DNA
-
 Aquifex aeolicus AMP + diphosphate + ?
-
 ?
6.5.1.2 ATP + nicked DNA
-
 Aquifex aeolicus ?
-
 ?
6.5.1.2 NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m activity depends on the base pairs on both sides of the nick, T/A Aquifex aeolicus 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 aeolicus AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
-
 ?

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
6.5.1.2 0.033
-
 nicked DNA pH 7.6, 65°C Aquifex aeolicus

pH Optimum

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

Cofactor

EC Number Cofactor Comment Organism Structure
6.5.1.2 NAD+ required, highest activity at 0.5 mM Aquifex aeolicus