Application | Comment | Organism |
---|---|---|
molecular biology | the H633R mutant DNA polymerase may be useful in high-fidelity DNA amplification and various PCR-based applications | Thermococcus waiotapuensis |
Cloned (Comment) | Organism |
---|---|
overexpression of the wild-type and mutant enzymes in Escherichia coli | Thermococcus waiotapuensis |
Protein Variants | Comment | Organism |
---|---|---|
H633D | DNA polymerase activity of the mutant enzyme is higher than the polymerase activity of the wild-type enzyme. PCR efficiency of the H633D mutant is higher than that of the N565K mutant enzyme | Thermococcus waiotapuensis |
H633R | significantly improved polymerase function compared to wild-type enzyme in terms of processivity (2-fold), extension rate (1.5fold) and PCR efficiency. The kcat value of the Twa H633R mutant is similar to that of wild-type, but the Km of the Twa H633R mutant is about 1.6fold lower than that of the wild-type | Thermococcus waiotapuensis |
N565K | DNA polymerase activity of the mutant enzyme is higher than the polymerase activity of the wild-type enzyme. PCR efficiency of the H633D mutant is higher than that of the N565K mutant enzyme | Thermococcus waiotapuensis |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0000066 | - |
DNAn | pH 8.6, 75°C, mM of template, in the presence of an excess of annealed primer, mutant enzyme H633R | Thermococcus waiotapuensis | |
0.0000104 | - |
DNAn | pH 8.6, 75°C, mM of template, in the presence of an excess of annealed prime, wild-type enzyme | Thermococcus waiotapuensis | |
0.003 | - |
deoxynucleoside triphosphate | pH 8.6, 75°C, mM of each nucleotide in an equimolar mixture of the four nucleotides, mutant enzyme H633R | Thermococcus waiotapuensis | |
0.003 | - |
deoxynucleoside triphosphate | pH 8.6, 75°C, mM of each nucleotide in an equimolar mixture of the four nucleotides, wild-type enzyme | Thermococcus waiotapuensis |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Thermococcus waiotapuensis | H9CW54 | - |
- |
Purification (Comment) | Organism |
---|---|
- |
Thermococcus waiotapuensis |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
deoxynucleoside triphosphate + DNAn | - |
Thermococcus waiotapuensis | diphosphate + DNAn+1 | - |
? |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
75 | - |
assay at | Thermococcus waiotapuensis |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
2.4 | - |
DNAn | pH 8.6, 75°C, mM of template, in the presence of an excess of annealed primer, mutant enzyme H633R | Thermococcus waiotapuensis | |
2.6 | - |
DNAn | pH 8.6, 75°C, mM of template, in the presence of an excess of annealed primer, wild-type enzyme | Thermococcus waiotapuensis | |
3.9 | - |
deoxynucleoside triphosphate | pH 8.6, 75°C, mM of each nucleotide in an equimolar mixture of the four nucleotides, wild-type enzyme | Thermococcus waiotapuensis | |
4.2 | - |
deoxynucleoside triphosphate | pH 8.6, 75°C, mM of each nucleotide in an equimolar mixture of the four nucleotides, mutant enzyme H633R | Thermococcus waiotapuensis |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.5 | - |
assay at | Thermococcus waiotapuensis |