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Literature summary for 2.7.7.6 extracted from
- Application of Escherichia coli phage K1E DNA-dependent RNA polymerase for in vitro RNA synthesis and in vivo protein production in Bacillus megaterium (2010), Appl. Microbiol. Biotechnol., 88, 529-539.
Activating Compound
| Activating Compound | Comment | Organism | Structure |
|---|---|---|---|
| spermidine | - |
Vectrevirus K1E |  |
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| gene k1ep, expression of His6-tagged K1E in Escherichia coli strain bL21(DE3), recombinant expression of K1E RNA polymerase in Bacillus megaterium strain MS941, a nprM deletion mutant of DSM319, using K1E promoters or SP6 promoter and a PEG-mediated transformation, method development and evaluation. The T7 RNAP promoter is not recognized at all | Vectrevirus K1E |
Metals/Ions
| Metals/Ions | Comment | Organism | Structure |
|---|---|---|---|
| Mg2+ | essentially required, best at 10 mM | Vectrevirus K1E | |
Molecular Weight [Da]
| Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
|---|---|---|---|
| 100000 | - |
1 * 100000, SDS-PAGE | Vectrevirus K1E |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Vectrevirus K1E | Q2WC24 | gene k1ep | - |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| recombinant His6-tagged K1E from Escherichia coli strain BL21(DE3) by nickel affinity chromatgraphy, dialysis and ultrafiltration | Vectrevirus K1E |
Specific Activity [micromol/min/mg]
| Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
|---|---|---|---|
| 4600 | - |
purified recombinant His6-tagged enzyme, pH 7.9, 37°C | Vectrevirus K1E |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| rNTP + RNAn | - |
Vectrevirus K1E | diphosphate + RNAn+1 | - |
? | |
Subunits
| Subunits | Comment | Organism |
|---|---|---|
| monomer | 1 * 100000, SDS-PAGE | Vectrevirus K1E |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| DNA-dependent RNA polymerase | - |
Vectrevirus K1E |
| DNA-dependent RNAP | - |
Vectrevirus K1E |
| K1E RNAP | - |
Vectrevirus K1E |
| RNAP | - |
Vectrevirus K1E |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 37 | 40 | - |
Vectrevirus K1E |
Temperature Range [°C]
| Temperature Minimum [°C] | Temperature Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 25 | 50 | activity range, profile, overview | Vectrevirus K1E |
Temperature Stability [°C]
| Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 25 | - |
half-life time of K1E RNAP stored in buffer containing 20 mM Na3PO4, 100 mM NaCl, pH 7.7, 1 mM EDTA, 10 mM DTT, and 0.1% Triton X-100, supplemented with 50% glycerol, is 87 h | Vectrevirus K1E |
| 50 | - |
half-life time of the K1E RNAP is 16.5 sec | Vectrevirus K1E |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 7.9 | - |
- |
Vectrevirus K1E |
pH Range
| pH Minimum | pH Maximum | Comment | Organism |
|---|---|---|---|
| 7.5 | 9 | 80% of maximall activity within this range | Vectrevirus K1E |
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