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Literature summary extracted from

  • Kim, J.H.; Lee, K.K.; Sun, Y.; Seo, G.J.; Cho, S.S.; Kwon, S.H.; Kwon, S.T.
    Broad nucleotide cofactor specificity of DNA ligase from the hyperthermophilic crenarchaeon Hyperthermus butylicus and its evolutionary significance (2013), Extremophiles, 17, 515-522.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
6.5.1.1 Hbu DNA ligase gene is expressed under control of the T7lac promoter of pTARG in Escherichia coli BL21-Codon-Plus(DE3)-RIL Hyperthermus butylicus
6.5.1.7 expressed under control of the T7lac promoter of pTARG in Escherichia coli BL21-Codon-Plus(DE3)-RIL Hyperthermus butylicus

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
6.5.1.1 Mg2+ required Hyperthermus butylicus
6.5.1.1 Mn2+ required Hyperthermus butylicus
6.5.1.7 Mg2+ the activity is highly dependent on MgCl2 or MnCl2 with maximal activity above 5 mM MgCl2 and 2 mM MnCl2 Hyperthermus butylicus
6.5.1.7 Mn2+ the activity is highly dependent on MgCl2 or MnCl2 with maximal activity above 5 mM MgCl2 and 2 mM MnCl2 Hyperthermus butylicus

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
6.5.1.1 68000
-
 SDS-PAGE Hyperthermus butylicus
6.5.1.7 68000
-
 x * 68000, SDS-PAGE Hyperthermus butylicus

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
6.5.1.7 ADP + (deoxyribonucleotide)n + (deoxyribonucleotide)m Hyperthermus butylicus the multiple cofactor specificity of the the enzyme may possibly be attributed to the ease of ATP decomposition at the high temperatures at which hyperthermophiles grow AMP + phosphate + (deoxyribonucleotide)n+m
-
 ?
6.5.1.7 ATP + (deoxyribonucleotide)n + (deoxyribonucleotide)m Hyperthermus butylicus the multiple cofactor specificity of the the enzyme may possibly be attributed to the ease of ATP decomposition at the high temperatures at which hyperthermophiles grow AMP + diphosphate + (deoxyribonucleotide)n+m
-
 ?
6.5.1.7 ATP + (deoxyribonucleotide)n + (deoxyribonucleotide)m Hyperthermus butylicus DSM Z 5456 the multiple cofactor specificity of the the enzyme may possibly be attributed to the ease of ATP decomposition at the high temperatures at which hyperthermophiles grow AMP + diphosphate + (deoxyribonucleotide)n+m
-
 ?
6.5.1.7 GTP + (deoxyribonucleotide)n + (deoxyribonucleotide)m Hyperthermus butylicus the multiple cofactor specificity of the the enzyme may possibly be attributed to the ease of ATP decomposition at the high temperatures at which hyperthermophiles grow GMP + diphosphate + (deoxyribonucleotide)n+m
-
 ?
6.5.1.7 GTP + (deoxyribonucleotide)n + (deoxyribonucleotide)m Hyperthermus butylicus DSM Z 5456 the multiple cofactor specificity of the the enzyme may possibly be attributed to the ease of ATP decomposition at the high temperatures at which hyperthermophiles grow GMP + diphosphate + (deoxyribonucleotide)n+m
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
6.5.1.1 Hyperthermus butylicus
-
-
-
6.5.1.7 Hyperthermus butylicus
-
-
-
6.5.1.7 Hyperthermus butylicus DSM Z 5456
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
6.5.1.1 purified using the IMPACTTM-CN system (intein-mediated purification with an affinity chitin-binding tag) and cation-ion (Arg-tag) chromatography Hyperthermus butylicus
6.5.1.7
-
 Hyperthermus butylicus

Reaction

EC Number Reaction Comment Organism Reaction ID
6.5.1.7 ADP + (deoxyribonucleotide)n-3'-hydroxyl + 5'-phospho-(deoxyribonucleotide)m = (deoxyribonucleotide)n+m + AMP + phosphate (2) Hyperthermus butylicus
a
6.5.1.7 ATP + (deoxyribonucleotide)n-3'-hydroxyl + 5'-phospho-(deoxyribonucleotide)m = (deoxyribonucleotide)n+m + AMP + diphosphate (1) Hyperthermus butylicus
a
6.5.1.7 GTP + (deoxyribonucleotide)n-3'-hydroxyl + 5'-phospho-(deoxyribonucleotide)m = (deoxyribonucleotide)n+m + GMP + diphosphate (3) Hyperthermus butylicus
a

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
6.5.1.7 ADP + (deoxyribonucleotide)n + (deoxyribonucleotide)m the multiple cofactor specificity of the the enzyme may possibly be attributed to the ease of ATP decomposition at the high temperatures at which hyperthermophiles grow Hyperthermus butylicus AMP + phosphate + (deoxyribonucleotide)n+m
-
 ?
6.5.1.7 ADP + (deoxyribonucleotide)n + (deoxyribonucleotide)m 95% nick-closing activity when compared with ATP Hyperthermus butylicus AMP + phosphate + (deoxyribonucleotide)n+m
-
 ?
6.5.1.7 ATP + (deoxyribonucleotide)n + (deoxyribonucleotide)m
-
 Hyperthermus butylicus AMP + diphosphate + (deoxyribonucleotide)n+m
-
 ?
6.5.1.7 ATP + (deoxyribonucleotide)n + (deoxyribonucleotide)m the multiple cofactor specificity of the the enzyme may possibly be attributed to the ease of ATP decomposition at the high temperatures at which hyperthermophiles grow Hyperthermus butylicus AMP + diphosphate + (deoxyribonucleotide)n+m
-
 ?
6.5.1.7 ATP + (deoxyribonucleotide)n + (deoxyribonucleotide)m
-
 Hyperthermus butylicus DSM Z 5456 AMP + diphosphate + (deoxyribonucleotide)n+m
-
 ?
6.5.1.7 ATP + (deoxyribonucleotide)n + (deoxyribonucleotide)m the multiple cofactor specificity of the the enzyme may possibly be attributed to the ease of ATP decomposition at the high temperatures at which hyperthermophiles grow Hyperthermus butylicus DSM Z 5456 AMP + diphosphate + (deoxyribonucleotide)n+m
-
 ?
6.5.1.7 GTP + (deoxyribonucleotide)n + (deoxyribonucleotide)m the multiple cofactor specificity of the the enzyme may possibly be attributed to the ease of ATP decomposition at the high temperatures at which hyperthermophiles grow Hyperthermus butylicus GMP + diphosphate + (deoxyribonucleotide)n+m
-
 ?
6.5.1.7 GTP + (deoxyribonucleotide)n + (deoxyribonucleotide)m 86% nick-closing activity when compared with ATP Hyperthermus butylicus GMP + diphosphate + (deoxyribonucleotide)n+m
-
 ?
6.5.1.7 GTP + (deoxyribonucleotide)n + (deoxyribonucleotide)m the multiple cofactor specificity of the the enzyme may possibly be attributed to the ease of ATP decomposition at the high temperatures at which hyperthermophiles grow Hyperthermus butylicus DSM Z 5456 GMP + diphosphate + (deoxyribonucleotide)n+m
-
 ?
6.5.1.7 GTP + (deoxyribonucleotide)n + (deoxyribonucleotide)m 86% nick-closing activity when compared with ATP Hyperthermus butylicus DSM Z 5456 GMP + diphosphate + (deoxyribonucleotide)n+m
-
 ?
6.5.1.7 additional information no activity with NAD+, CTP or TTP Hyperthermus butylicus ?
-
 ?
6.5.1.7 additional information no activity with NAD+, CTP or TTP Hyperthermus butylicus DSM Z 5456 ?
-
 ?

Subunits

EC Number Subunits Comment Organism
6.5.1.7 ? x * 68000, SDS-PAGE Hyperthermus butylicus

Synonyms

EC Number Synonyms Comment Organism
6.5.1.1 Hbu DNA ligase
-
 Hyperthermus butylicus
6.5.1.7 Hbu DNA ligase
-
 Hyperthermus butylicus

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
6.5.1.1 75
-
-
 Hyperthermus butylicus
6.5.1.7 75
-
-
 Hyperthermus butylicus

Temperature Range [°C]

EC Number Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
6.5.1.7 60 80 60°C: about 55% of maximal activity, 80°C: about 60% of maximal activity Hyperthermus butylicus

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
6.5.1.1 94
-
 Hbu DNA ligase exhibits about 50% of the original activity for 2.3 h at temperatures up to 94 °C Hyperthermus butylicus
6.5.1.1 99
-
 Hbu DNA ligase exhibtis a 1.7 h half life at 99 °C Hyperthermus butylicus
6.5.1.7 94
-
 2.3 h, about 50% loss of activity Hyperthermus butylicus
6.5.1.7 99
-
 half-life: 1.7 h Hyperthermus butylicus

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
6.5.1.1 8
-
-
 Hyperthermus butylicus
6.5.1.7 8
-
 in Tris-HCl buffer Hyperthermus butylicus

pH Range

EC Number pH Minimum pH Maximum Comment Organism
6.5.1.7 6.5 9 pH 6.5: about 90% of maximal activity, 9.0: about 95% of maximal activity Hyperthermus butylicus

Cofactor

EC Number Cofactor Comment Organism Structure
6.5.1.1 ADP
-
 Hyperthermus butylicus
6.5.1.1 ATP
-
 Hyperthermus butylicus
6.5.1.1 GTP
-
 Hyperthermus butylicus
6.5.1.1 additional information enzyme shows a broad nucleotide cofactor specificity Hyperthermus butylicus