Activating Compound | Comment | Organism | Structure |
---|---|---|---|
GTP | activation of wild-type and mutant P131D, maximal at 0.08 mM | Escherichia coli |
Cloned (Comment) | Organism |
---|---|
overexpression of wild-type and mutant P131D in Escherichia coli | Escherichia coli |
Protein Variants | Comment | Organism |
---|---|---|
P131D | site-directed mutagenesis, 2-step mutagenic PCR, exchange of proline in 5-phosphoribose 1-diphosphate binding site, 50-60fold reduction of catalytic rate in both reaction directions, about 100fold increase in KM for uracil, strongly reduced Km for 5-phosphoribose 1-diphosphate | Escherichia coli |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | Km-value for 5-phosphoribose 1-diphosphate of the mutant P131D is very low and difficult to determine | Escherichia coli | |
0.00063 | - |
Uracil | recombinant wild-type | Escherichia coli | |
0.06 | - |
Uracil | recombinant mutant P131D | Escherichia coli | |
0.073 | - |
5-phospho-alpha-D-ribose 1-diphosphate | recombinant wild-type | Escherichia coli |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | - |
Escherichia coli |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Escherichia coli | - |
gene upp | - |
Purification (Comment) | Organism |
---|---|
recombinant wild-type and mutant P131D from overexpressing Escherichia coli | Escherichia coli |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
UMP + diphosphate = uracil + 5-phospho-alpha-D-ribose 1-diphosphate | sequential reaction mechanism | Escherichia coli | |
UMP + diphosphate = uracil + 5-phospho-alpha-D-ribose 1-diphosphate | Pro131 is of little importance for 5-phospho-alpha-D-ribose 1-diphosphate binding, but critical for binding of uracil to the enzyme-5-phospho-alpha-D-ribose 1-diphosphate complex | Escherichia coli |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
additional information | - |
- |
Escherichia coli |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
uracil + 5-phospho-alpha-D-ribose 1-diphosphate | 5-phospho-alpha-D-ribose 1-diphosphate binding site | Escherichia coli | UMP + diphosphate | - |
r |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
assay at | Escherichia coli |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
56 | 57 | 10 min, 50% loss of activity, recombinant wild-type and mutant, scanning calorimetry | Escherichia coli |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
8.5 | - |
assay at | Escherichia coli |