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Literature summary for 3.1.26.4 extracted from
- Gene cloning and biochemical characterizations of thermostable ribonuclease HIII from Bacillus stearothermophilus (2004), Biosci. Biotechnol. Biochem., 68, 2138-2147.
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| gene rnhC, DNA and amino acid sequence determination and analysis, overexpression of the soluble enzyme in Escherichia coli strain MIC2067(DE3) | Geobacillus stearothermophilus |
Inhibitors
| Inhibitors | Comment | Organism | Structure |
|---|---|---|---|
| Co2+ | over 95% inhibition at 5 mM and below | Geobacillus stearothermophilus |  |
| Mn2+ | 40% inhibition at 100 mM | Geobacillus stearothermophilus | |
| Ni2+ | nearly complete inhibition at 5 mM and below | Geobacillus stearothermophilus | |
Metals/Ions
| Metals/Ions | Comment | Organism | Structure |
|---|---|---|---|
| KCl | highly activating, best at 100-200 mM salt, KCl is preferred | Geobacillus stearothermophilus | |
| Mg2+ | required, highest activity at 50 mM | Geobacillus stearothermophilus | |
| additional information | Mg2+ cannot be substituted by Co2+ and Ni2+, and only partially by Mn2+ | Geobacillus stearothermophilus | |
| NaCl | activating, best at 100-200 mM salt | Geobacillus stearothermophilus | |
Molecular Weight [Da]
| Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
|---|---|---|---|
| 33734 | - |
1 * 33734, amino acid sequence calculation, 1 * 36000, recombinant enzyme, SDS-PAGE | Geobacillus stearothermophilus |
| 36000 | - |
recombinant enzyme, gel filtration | Geobacillus stearothermophilus |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Geobacillus stearothermophilus | Q6L6Q4 | gene rnhC | - |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| recombinant enzyme to homogeneity from Escherichia coli strain MIC2067(DE3) by ammonium sulfate precipitation, heparin affinity chromatography, and gel filtration | Geobacillus stearothermophilus |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| 12 basepair DNA-DNA duplex + H2O | oligomeric substrate, cleavage at multiple sites, product identification | Geobacillus stearothermophilus | ? | - |
? | |
| 29 basepair DNA-RNA-DNA/DNA + H2O | oligomeric substrate, cleavage mainly in the middle of the tetraribonucleotide, product identification | Geobacillus stearothermophilus | ? | - |
? | |
| M13 DNA-RNA hybrid + H2O | - |
Geobacillus stearothermophilus | ? | - |
? | |
Subunits
| Subunits | Comment | Organism |
|---|---|---|
| monomer | 1 * 33734, amino acid sequence calculation, 1 * 36000, recombinant enzyme, SDS-PAGE | Geobacillus stearothermophilus |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| ribonuclease HIII | - |
Geobacillus stearothermophilus |
| RNase HIII | - |
Geobacillus stearothermophilus |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 70 | - |
above | Geobacillus stearothermophilus |
Temperature Range [°C]
| Temperature Minimum [°C] | Temperature Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 30 | 70 | and above, temperature profile | Geobacillus stearothermophilus |
Temperature Stability [°C]
| Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 55 | - |
half-life 10 min | Geobacillus stearothermophilus |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 8.5 | 9 | - |
Geobacillus stearothermophilus |
pI Value
| Organism | Comment | pI Value Maximum | pI Value |
|---|---|---|---|
| Geobacillus stearothermophilus | sequence calculation | - |
8.6 |
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