Cloned (Comment) | Organism |
---|---|
gene rnhA, overexpression in Escherichia coli MIC3001 of wild-type and mutant enzymes, complementation of enzyme-dependent temperature-sensitive growth phenotype of the Escherichia coli host strain by expression of the wild-type enzyme, but only marginally by the mutant D134H | Thermus thermophilus |
Protein Variants | Comment | Organism |
---|---|---|
A12S | random mutagenesis, roughly cumulative effect on enzyme activity, increased kcat/Km value compared to the wild-type enzyme | Thermus thermophilus |
A12S/K75M/A77P | site-directed mutagenesis, 40fold increased kcat/Km value compared to the wild-type enzyme, thermal stability similar to the wild-type enzyme | Thermus thermophilus |
A77P | random mutagenesis, roughly cumulative effect on enzyme activity, increased kcat/Km value compared to the wild-type enzyme | Thermus thermophilus |
D134H | site-directed mutagenesis, active site mutant, highly reduced catalytic activity | Thermus thermophilus |
D134N | site-directed mutagenesis, active site mutant, similar to the wild-type enzyme | Thermus thermophilus |
K75M | random mutagenesis, roughly cumulative effect on enzyme activity, increased kcat/Km value compared to the wild-type enzyme | Thermus thermophilus |
additional information | isolation of a mutant enzyme with enhanced activity at moderate temperatures, genetic method development | Thermus thermophilus |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | thermodynamics of wild-type and mutant enzymes | Thermus thermophilus | |
0.0011 | - |
DNA-RNA-DNA hybrid | mutant A12S/K75M and A12S/K75M/A77P, pH 8.0, 30°C | Thermus thermophilus | |
0.0013 | - |
DNA-RNA-DNA hybrid | mutant A12S/A77P, pH 8.0, 30°C | Thermus thermophilus | |
0.0015 | - |
DNA-RNA-DNA hybrid | mutant A12S, pH 8.0, 30°C | Thermus thermophilus | |
0.002 | - |
DNA-RNA-DNA hybrid | mutant K75M/A77P, pH 8.0, 30°C | Thermus thermophilus | |
0.0033 | - |
DNA-RNA-DNA hybrid | mutant D134H and A77P, pH 8.0, 30°C | Thermus thermophilus | |
0.0061 | - |
DNA-RNA-DNA hybrid | mutant K75M, pH 8.0, 30°C | Thermus thermophilus | |
0.0078 | - |
DNA-RNA-DNA hybrid | wild-type enzyme, pH 8.0, 30°C | Thermus thermophilus |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | - |
Thermus thermophilus |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Thermus thermophilus | - |
type I enzyme, gene rnhA | - |
Thermus thermophilus HB8 / ATCC 27634 / DSM 579 | - |
type I enzyme, gene rnhA | - |
Purification (Comment) | Organism |
---|---|
recombinant wild-type and mutant enzymes from Escherichia coli | Thermus thermophilus |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
additional information | - |
activity of wild-type and mutant enzymes at different temperatures | Thermus thermophilus |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
DNA-RNA-DNA hybrid + H2O | specific hydrolysis of the RNA strand of the hybrid | Thermus thermophilus | ? | - |
? | |
DNA-RNA-DNA hybrid + H2O | specific hydrolysis of the RNA strand of the hybrid | Thermus thermophilus HB8 / ATCC 27634 / DSM 579 | ? | - |
? |
Synonyms | Comment | Organism |
---|---|---|
ribonuclease HI | - |
Thermus thermophilus |
RNase HI | - |
Thermus thermophilus |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
30 | - |
assay at | Thermus thermophilus |
60 | - |
assay at | Thermus thermophilus |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.003 | - |
DNA-RNA-DNA hybrid | mutant D134H, pH 8.0, 30°C | Thermus thermophilus | |
0.075 | - |
DNA-RNA-DNA hybrid | mutant A12S, pH 8.0, 30°C | Thermus thermophilus | |
0.082 | - |
DNA-RNA-DNA hybrid | wild-type enzyme, pH 8.0, 30°C | Thermus thermophilus | |
0.122 | - |
DNA-RNA-DNA hybrid | mutant A77P, pH 8.0, 30°C | Thermus thermophilus | |
0.133 | - |
DNA-RNA-DNA hybrid | mutant K75M, pH 8.0, 30°C | Thermus thermophilus | |
0.2 | - |
DNA-RNA-DNA hybrid | mutant A12S/K75M, pH 8.0, 30°C | Thermus thermophilus | |
0.25 | - |
DNA-RNA-DNA hybrid | mutant A12S/A77P, pH 8.0, 30°C | Thermus thermophilus | |
0.33 | - |
DNA-RNA-DNA hybrid | mutant K75M/A77P, pH 8.0, 30°C | Thermus thermophilus | |
0.45 | - |
DNA-RNA-DNA hybrid | mutant A12S/K75M/A77P, pH 8.0, 30°C | Thermus thermophilus |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
8 | - |
assay at | Thermus thermophilus |