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Literature summary for 2.7.7.7 extracted from
- 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
| Activating Compound | Comment | Organism | Structure |
|---|---|---|---|
| (NH4)2SO4 | optimal concentration of 20 mM, inhibition below and above 20 mM | Thermococcus pacificus |  |
Application
| Application | Comment | Organism |
|---|---|---|
| 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)
| Cloned (Comment) | Organism |
|---|---|
| expression in Escherichia coli | Thermococcus pacificus |
Protein Variants
| Protein Variants | Comment | Organism |
|---|---|---|
| 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
| Inhibitors | Comment | Organism | Structure |
|---|---|---|---|
| (NH4)2SO4 | optimal concentration of 20 mM, inhibition below and above 20 mM | Thermococcus pacificus | |
| KCl | - |
Thermococcus pacificus | |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 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 | |
| 0.0000057 | - |
DNAn | pH 7.5, 75°C, template in the presence of an excess of annealed primer, unmodified Tpa DNA polymerase | Thermococcus pacificus | |
| 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 | |
| 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
| Metals/Ions | Comment | Organism | Structure |
|---|---|---|---|
| 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]
| Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
|---|---|---|---|
| 90000 | - |
x * 90000, SDS-PAGE | Thermococcus pacificus |
| 90054 | - |
x * 90054, calculated from sequence | Thermococcus pacificus |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Thermococcus pacificus | - |
- |
- |
| Thermococcus pacificus DSM 10394 | - |
- |
- |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
- |
Thermococcus pacificus |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| deoxynucleoside triphosphate + DNAn | - |
Thermococcus pacificus | diphosphate + DNAn+1 | - |
? | |
| deoxynucleoside triphosphate + DNAn | - |
Thermococcus pacificus DSM 10394 | diphosphate + DNAn+1 | - |
? | |
Subunits
| Subunits | Comment | Organism |
|---|---|---|
| ? | x * 90000, SDS-PAGE | Thermococcus pacificus |
| ? | x * 90054, calculated from sequence | Thermococcus pacificus |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 75 | - |
- |
Thermococcus pacificus |
Temperature Range [°C]
| Temperature Minimum [°C] | Temperature Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 65 | 90 | 65°C: about 40% of maximal activity, 90°C: about 50% of maximal activity | Thermococcus pacificus |
Temperature Stability [°C]
| Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 99 | - |
half-life: 3 h | Thermococcus pacificus |
Turnover Number [1/s]
| Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 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 | |
| 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 | |
| 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 | |
| 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 | |
| 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
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 7.5 | - |
- |
Thermococcus pacificus |
pH Range
| pH Minimum | pH Maximum | Comment | Organism |
|---|---|---|---|
| 6.5 | 8.5 | pH 6.5: about 50% of maximal activity, pH 8.5: about 40% of maximal activity | Thermococcus pacificus |
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