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Literature summary extracted from

  • Lee, J.I.; Cho, S.S.; Kil, E.-J.; Kwon, S.-T.
    Characterization and PCR application of a thermostable DNA polymerase from Thermococcus pacificus (2010), Enzyme Microb. Technol., 47, 147-152.
No PubMed abstract available

Activating Compound

EC Number Activating Compound Comment Organism Structure
2.7.7.7 (NH4)2SO4 optimal concentration of 20 mM, inhibition below and above 20 mM Thermococcus pacificus

Application

EC Number Application Comment Organism
2.7.7.7 molecular biology generation of a unique one enzyme system with high fidelity to allow highly accurate and efficient amplification of DNA targets using polymerase chain reaction by fusing Sso7d protein to Tpa DNA polymerase Thermococcus pacificus

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.7.7.7 expression in Escherichia coli Thermococcus pacificus

Protein Variants

EC Number Protein Variants Comment Organism
2.7.7.7 additional information Tpa-S DNA polymerase (the fusion of the Sso7d protein to the C-terminus of Tpa DNA polymerase) improves the performance of the Tpa DNA polymerase to make it suitable for long and rapid polymerase chain reaction Thermococcus pacificus

Inhibitors

EC Number Inhibitors Comment Organism Structure
2.7.7.7 (NH4)2SO4 optimal concentration of 20 mM, inhibition below and above 20 mM Thermococcus pacificus
2.7.7.7 KCl
-
 Thermococcus pacificus

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.7.7.7 0.0000032
-
 DNAn pH 7.5, 75°C, template in the presence of an excess of annealed primer, fusion protein of the Sso7d protein to the C-terminus of Tpa DNA polymerase Thermococcus pacificus
2.7.7.7 0.0000057
-
 DNAn pH 7.5, 75°C, template in the presence of an excess of annealed primer, unmodified Tpa DNA polymerase Thermococcus pacificus
2.7.7.7 0.17
-
 deoxynucleoside triphosphate pH 7.5, 75°C, KM-value of each nucleotide in an equimolar mixture of the four nucleotides, fusion protein of the Sso7d protein to the C-terminus of Tpa DNA polymerase Thermococcus pacificus
2.7.7.7 0.64
-
 deoxynucleoside triphosphate pH 7.5, 75°C, KM-value of each nucleotide in an equimolar mixture of the four nucleotides, unmodified Tpa DNA polymerase Thermococcus pacificus

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
2.7.7.7 Mg2+ highly dependent on MgCl2, with maximal activity at 12 mM MgCl2 and no detectable activity in the absence of MgCl2 Thermococcus pacificus

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
2.7.7.7 90000
-
 x * 90000, SDS-PAGE Thermococcus pacificus
2.7.7.7 90054
-
 x * 90054, calculated from sequence Thermococcus pacificus

Organism

EC Number Organism UniProt Comment Textmining
2.7.7.7 Thermococcus pacificus
-
-
-
2.7.7.7 Thermococcus pacificus DSM 10394
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.7.7.7
-
 Thermococcus pacificus

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.7.7.7 deoxynucleoside triphosphate + DNAn
-
 Thermococcus pacificus diphosphate + DNAn+1
-
 ?
2.7.7.7 deoxynucleoside triphosphate + DNAn
-
 Thermococcus pacificus DSM 10394 diphosphate + DNAn+1
-
 ?

Subunits

EC Number Subunits Comment Organism
2.7.7.7 ? x * 90000, SDS-PAGE Thermococcus pacificus
2.7.7.7 ? x * 90054, calculated from sequence Thermococcus pacificus

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
2.7.7.7 75
-
-
 Thermococcus pacificus

Temperature Range [°C]

EC Number Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
2.7.7.7 65 90 65°C: about 40% of maximal activity, 90°C: about 50% of maximal activity Thermococcus pacificus

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
2.7.7.7 99
-
 half-life: 3 h Thermococcus pacificus

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
2.7.7.7 additional information
-
 additional information Tpa-S DNA polymerase (the fusion of the Sso7d protein to the C-terminus of Tpa DNA polymerase) has nearly an identical turnover rate for DNA with the unmodified Tpa DNA polymerase. However, Tpa-S DNA polymerase has a somewhat higher (about 1.26fold) turnover rate in nucleotide incorporation than Tpa DNA polymerase Thermococcus pacificus
2.7.7.7 12.9
-
 DNAn pH 7.5, 75°C, mol of template, in the presence of an excess of annealed primer, fusion protein of the Sso7d protein to the C-terminus of Tpa DNA polymerase Thermococcus pacificus
2.7.7.7 13.6
-
 DNAn pH 7.5, 75°C, mol of template, in the presence of an excess of annealed primer, unmodified Tpa DNA polymerase Thermococcus pacificus
2.7.7.7 92
-
 deoxynucleoside triphosphate pH 7.5, 75°C, turnover number of each nucleotide in an equimolar mixture of the four nucleotides, unmodified Tpa DNA polymerase Thermococcus pacificus
2.7.7.7 116
-
 deoxynucleoside triphosphate pH 7.5, 75°C, turnover number of each nucleotide in an equimolar mixture of the four nucleotides, fusion protein of the Sso7d protein to the C-terminus of Tpa DNA polymerase Thermococcus pacificus

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
2.7.7.7 7.5
-
-
 Thermococcus pacificus

pH Range

EC Number pH Minimum pH Maximum Comment Organism
2.7.7.7 6.5 8.5 pH 6.5: about 50% of maximal activity, pH 8.5: about 40% of maximal activity Thermococcus pacificus