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Literature summary for 2.7.7.7 extracted from
- Temperature dependence of accuracy of DNA polymerase I from Geobacillus anatolicus (2012), Biochimie, 94, 1968-1973.
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| gene polA, Klenow-like DNA polymerase I, DNA and amino acid sequence determination and analysis, expression of His-tagged protein in Escherichia coli strain JM109(DE3) | Geobacillus anatolicus |
Protein Variants
| Protein Variants | Comment | Organism |
|---|---|---|
| D424A | a proofreading deficient mutant of the Klenow fragment | Geobacillus anatolicus |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| additional information | - |
additional information | in pre-steady-state kinetic measurements, when temperature is raised from 22°C to 50°C, the rate (kpol) for cognate dTTP and non-cognate dATP nucleotide incorporations increases 6 and 4fold, respectively, whereas the Kd for both nucleotide incorporations changes only slightly. Single turnover kinetics | Geobacillus anatolicus | |
| 0.0314 | - |
dTTP | pH 8.0, 22°C, recombinant enzyme | Geobacillus anatolicus |  |
| 0.253 | - |
dATP | pH 8.0, 37°C, recombinant enzyme | Geobacillus anatolicus | |
| 0.274 | - |
dATP | pH 8.0, 50°C, recombinant enzyme | Geobacillus anatolicus | |
| 0.316 | - |
dTTP | pH 8.0, 22°C, recombinant enzyme | Geobacillus anatolicus | |
| 38.1 | - |
dTTP | pH 8.0, 37°C, recombinant enzyme | Geobacillus anatolicus | |
| 47.4 | - |
dTTP | pH 8.0, 50°C, recombinant enzyme | Geobacillus anatolicus | |
Metals/Ions
| Metals/Ions | Comment | Organism | Structure |
|---|---|---|---|
| Mg2+ | required | Geobacillus anatolicus | |
Natural Substrates/ Products (Substrates)
| Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| deoxynucleoside triphosphate + DNAn | Geobacillus anatolicus | - |
diphosphate + DNAn+1 | - |
? | |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Geobacillus anatolicus | - |
- |
- |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| recombinant His-tagged Klenow-like DNA polymerase I from Escherichia coli strain JM109(DE3) by nickel affinity chromatography and heat treatment at 80°C for 30 min | Geobacillus anatolicus |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| dATP + DNAn | with labeled 20/33-mer primer-template duplex DNA | Geobacillus anatolicus | diphosphate + DNAn+1 | - |
? | |
| deoxxynucleoside triphosphate + DNAn | the phosphoryl transfer step may be rate limiting for the non-cognate nucleotide incorporation by the enzyme | Geobacillus anatolicus | diphosphate + DNAn+1 | - |
? | |
| deoxynucleoside triphosphate + DNAn | - |
Geobacillus anatolicus | diphosphate + DNAn+1 | - |
? | |
| dTTP + DNAn | with labeled 20/33-mer primer-template duplex DNA | Geobacillus anatolicus | diphosphate + DNAn+1 | - |
? | |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| DNA polymerase I | - |
Geobacillus anatolicus |
| Klenow-like DNA polymerase I | - |
Geobacillus anatolicus |
| polI | - |
Geobacillus anatolicus |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 22 | 50 | assay at | Geobacillus anatolicus |
Temperature Range [°C]
| Temperature Minimum [°C] | Temperature Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 22 | 72 | activity range, the error frequency of the enzyme does not change significantly when the temperature is raised from 22°C to 72°C | Geobacillus anatolicus |
Turnover Number [1/s]
| Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.096 | - |
dTTP | pH 8.0, 22°C, recombinant enzyme | Geobacillus anatolicus | |
| 0.185 | - |
dATP | pH 8.0, 37°C, recombinant enzyme | Geobacillus anatolicus | |
| 0.4 | - |
dATP | pH 8.0, 50°C, recombinant enzyme | Geobacillus anatolicus | |
| 25.8 | - |
dTTP | pH 8.0, 22°C, recombinant enzyme | Geobacillus anatolicus | |
| 75.6 | - |
dTTP | pH 8.0, 37°C, recombinant enzyme | Geobacillus anatolicus | |
| 175 | - |
dTTP | pH 8.0, 50°C, recombinant enzyme | Geobacillus anatolicus | |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 8 | - |
assay at | Geobacillus anatolicus |
General Information
| General Information | Comment | Organism |
|---|---|---|
| additional information | the error frequency of the enzyme does not change significantly when the temperature is raised from 22°C to 72°C | Geobacillus anatolicus |
kcat/KM [mM/s]
| kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.304 | - |
dTTP | pH 8.0, 22°C, recombinant enzyme | Geobacillus anatolicus | |
| 0.731 | - |
dATP | pH 8.0, 37°C, recombinant enzyme | Geobacillus anatolicus | |
| 1.46 | - |
dATP | pH 8.0, 50°C, recombinant enzyme | Geobacillus anatolicus | |
| 821.7 | - |
dTTP | pH 8.0, 22°C, recombinant enzyme | Geobacillus anatolicus | |
| 1984 | - |
dTTP | pH 8.0, 37°C, recombinant enzyme | Geobacillus anatolicus | |
| 3692 | - |
dTTP | pH 8.0, 50°C, recombinant enzyme | Geobacillus anatolicus | |
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