Cloned (Comment) | Organism |
---|---|
expression of the N-terminal part of the LF Bst polymerase gene in Escherichia coli strain BL21 | Geobacillus stearothermophilus |
Protein Variants | Comment | Organism |
---|---|---|
additional information | engineered DNA polymerases are obtained by the flexible attachment of helix-hairpin-helix DNA binding domains of topoisomerase V of Methanopyrus kandleri to catalytic domains of DNA polymerases, domain fusion using the Taq and Pfu DNA polymerases or the phi29 DNA polymerase. The chimeric polymerases retain high processivity at high concentrations of salts and other inhibitors of DNA synthesis, such as phenol, blood, and DNA intercalating dyes, domain attachment has a potential to greatly increase the thermostability of chimeric DNA polymerases | Methanopyrus kandleri |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
deoxynucleoside triphosphate + DNAn | Geobacillus stearothermophilus | - |
diphosphate + DNAn+1 | - |
? | |
deoxynucleoside triphosphate + DNAn | Methanopyrus kandleri | - |
diphosphate + DNAn+1 | - |
? | |
deoxynucleoside triphosphate + DNAn | Geobacillus stearothermophilus Donc | - |
diphosphate + DNAn+1 | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Geobacillus stearothermophilus | - |
ATCC12980D-5 | - |
Geobacillus stearothermophilus Donc | - |
ATCC12980D-5 | - |
Methanopyrus kandleri | - |
- |
- |
Purification (Comment) | Organism |
---|---|
recombinant N-terminal part of the LF Bst polymerase from Escherichia coli strain BL21 by anion exchange chromatography | Geobacillus stearothermophilus |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
deoxynucleoside triphosphate + DNAn | - |
Geobacillus stearothermophilus | diphosphate + DNAn+1 | - |
? | |
deoxynucleoside triphosphate + DNAn | - |
Methanopyrus kandleri | diphosphate + DNAn+1 | - |
? | |
deoxynucleoside triphosphate + DNAn | standard substrate PTJ1 and substrate PTJ2 | Geobacillus stearothermophilus | diphosphate + DNAn+1 | - |
? | |
deoxynucleoside triphosphate + DNAn | standard substrate PTJ1 and substrate PTJ2 | Methanopyrus kandleri | diphosphate + DNAn+1 | - |
? | |
deoxynucleoside triphosphate + DNAn | - |
Geobacillus stearothermophilus Donc | diphosphate + DNAn+1 | - |
? | |
deoxynucleoside triphosphate + DNAn | standard substrate PTJ1 and substrate PTJ2 | Geobacillus stearothermophilus Donc | diphosphate + DNAn+1 | - |
? |
Synonyms | Comment | Organism |
---|---|---|
Bst DNA polymerase | - |
Geobacillus stearothermophilus |
Mka polB | - |
Methanopyrus kandleri |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
25 | - |
assay at | Geobacillus stearothermophilus |
25 | - |
assay at | Methanopyrus kandleri |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
25 | 95 | thermostability and half-life of the LF Bst polymerase with the attached TopoV's HhH C2 domain increases almost 8fold, as compared to the Bst LF polymerase alone. The effect can be seen up to 95°C. The stabilizing effect of the C3 domain in N-TopoTaq completely disappears if K-Glu is replaced by 0.5 M NaCl, and the stability of N-TopoTaq does not differ from that of the catalytic domain alone, addition of 1 M betaine restores the stabilization provided by the C3 domain. Electrostatic enhancement of diffusion-controlled association. Thermal stabilization of chimeric polymerases occurs due to charge-independant interactions of the polymerase domain and HhH domains | Geobacillus stearothermophilus |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
8 | - |
assay at | Geobacillus stearothermophilus |
8 | - |
assay at | Methanopyrus kandleri |
General Information | Comment | Organism |
---|---|---|
evolution | the enzyme belongs to the DNA polymerase family B | Methanopyrus kandleri |