Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli JM83(pMLD58) | Escherichia coli |
Crystallization (Comment) | Organism |
---|---|
hanging-drop vapor diffusion technique | Escherichia coli |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
arsenate | increases storage stability | Escherichia coli | |
Ca2+ | increases storage stability | Escherichia coli | |
Cd2+ | abolishes the enzyme activity at 5mM | Escherichia coli | |
Mg2+ | increases storage stability | Escherichia coli | |
Mn2+ | increases storage stability | Escherichia coli | |
phosphate | increases storage stability | Escherichia coli | |
sulfate | increases storage stability | Escherichia coli | |
vanadate | abolishes the enzyme activity at 5mM | Escherichia coli | |
Zn2+ | abolishes the enzyme activity at 5mM | Escherichia coli |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
46840 | - |
mass spectrometry | Escherichia coli |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Escherichia coli | - |
- |
- |
Escherichia coli JM83(pMLD58) | - |
- |
- |
Purification (Comment) | Organism |
---|---|
method that includes DEAE-Trisacryl M and hydroxyapatite-Ultrogel chromatography. Large-scale purification using 2-mercaptoethanol as a reducing agent, results in 200 mg of purified enzyme from 54 l of culture, adducts with one or two molecules of 2-mercaptoethanol are formed. Replacement of 2-mercaptoethanol by dithiothreitol gave 162 mg of enzyme from 36 l of culture yielding no clusters with the reducing agent. | Escherichia coli |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
1.772 | - |
37°C | Escherichia coli |
Storage Stability | Organism |
---|---|
-20°C, 20 mM potassium phosphate, pH 7.0, 1 mM 2-mercaptoethanol, 0.1 mM MgCl2, several months, no loss of activity at high enzyme concentrations: 1-10 mg/ml | Escherichia coli |
-20°C, 20 mM potassium phosphate, pH 7.0, 1 mM dithiothreitol, 0.1 mM MgCl2, several months, no loss of activity at high enzyme concentrations: 1-10 mg/ml | Escherichia coli |
8°C, 20 mM potassium phosphate, pH 7.0, 1 mM 2-mercaptoethanol, 0.1 mM MgCl2, 1 week, 30% loss of activity | Escherichia coli |
8°C, 20 mM potassium phosphate, pH 7.0, 1 mM 2-mercaptoethanol, 0.1 mM MgCl2, sodium Hepes, 1 week, 35-43% loss of activity | Escherichia coli |
8°C, 20 mM potassium phosphate, pH 7.0, 1 mM dithiothreitol, 0.1 mM MgCl2, 1 week, 30% loss of activity | Escherichia coli |
8°C, 20 mM potassium phosphate, pH 7.0, 1 mM dithiothreitol, 0.1 mM MgCl2, sodium Hepes 1 week, 35-43% loss of activity | Escherichia coli |
8°C, 20 mM potassium phosphate, pH 8.0, 1 mM 2-mercaptoethanol, 0.1 mM MgCl2, sodium Hepes, 1 week, 35-43% loss of activity | Escherichia coli |
8°C, 20 mM potassium phosphate, pH 8.0, 1 mM dithiothreitol, 0.1 mM MgCl2, sodium Hepes 1 week, 35-43% loss of activity | Escherichia coli |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | turnover of 6.55 at saturation concentrations of all the substrates: 5 mM ATP, 0.25 mM UDP-N-acetylmuramoyl L-alanine, 0.55 mM D-glutamate | Escherichia coli |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
8.8 | - |
- |
Escherichia coli |