Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli BL21(DE3) cells | Escherichia coli |
Crystallization (Comment) | Organism |
---|---|
in complex with isochorismate and Mg2+, hanging drop vapor diffusion method, using 100 mM MES buffer pH 6.5, 12% PEG (w/v) 20000 | Escherichia coli |
Protein Variants | Comment | Organism |
---|---|---|
A303T | the kcat/Km (chorismate) for the A303T mutant is 130fold lower than that for wild type enzyme | Escherichia coli |
A303T/F327Y | the mutant shows no detectable activity | Escherichia coli |
A303T/F327Y/F359Q | the mutant shows no detectable activity | Escherichia coli |
A303T/F327Y/F359Q/I346L | the mutant shows no detectable activity | Escherichia coli |
A303T/F327Y/I346L | the mutant shows no detectable activity | Escherichia coli |
A303T/F359Q | the mutant shows no detectable activity | Escherichia coli |
F327Y | the mutant displays reduced activity compared to the wild type enzyme | Escherichia coli |
F327Y/F359Q | the mutant displays a reduced kcat/Km value compared to the wild type enzyme | Escherichia coli |
F327Y/F359Q/I346L | the mutant shows no detectable activity | Escherichia coli |
F359Q | the mutant displays a reduced kcat/Km value compared to the wild type enzyme | Escherichia coli |
F359Q | the mutation causes loss of catalytic activity | Escherichia coli |
F359Q/I346L | the mutant displays a reduced kcat/Km value compared to the wild type enzyme | Escherichia coli |
I346L | the EntC mutant has a 12fold lower kcat/Km (chorismate) than the wild type enzyme | Escherichia coli |
L304A | the mutation causes loss of catalytic activity | Escherichia coli |
L304A/F359Q | the mutant shows no detectable activity | Escherichia coli |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.007 | - |
chorismate | wild type enzyme, in 100 mM phosphate pH 7.0, 10 mM MgCl2 | Escherichia coli | |
0.04 | - |
chorismate | mutant enzyme F327Y/F359Q, in 100 mM phosphate pH 7.0, 10 mM MgCl2 | Escherichia coli | |
0.066 | - |
chorismate | mutant enzyme F359Q, in 100 mM phosphate pH 7.0, 10 mM MgCl2 | Escherichia coli | |
0.076 | - |
chorismate | mutant enzyme F327Y, in 100 mM phosphate pH 7.0, 10 mM MgCl2 | Escherichia coli | |
0.076 | - |
chorismate | mutant enzyme I346L, in 100 mM phosphate pH 7.0, 10 mM MgCl2 | Escherichia coli | |
0.103 | - |
chorismate | mutant enzyme A303T, in 100 mM phosphate pH 7.0, 10 mM MgCl2 | Escherichia coli | |
0.261 | - |
chorismate | mutant enzyme F359Q/I346L, in 100 mM phosphate pH 7.0, 10 mM MgCl2 | Escherichia coli | |
0.466 | - |
chorismate | mutant enzyme F327Y/I346L, in 100 mM phosphate pH 7.0, 10 mM MgCl2 | Escherichia coli |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | the enzyme contains Mg2+ | Escherichia coli |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Escherichia coli | P0AEJ2 | - |
- |
Purification (Comment) | Organism |
---|---|
Ni2+-charged HiTrap chelating agarose column chromatography and Superdex 75 gel filtration | Escherichia coli |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
Chorismate | - |
Escherichia coli | Isochorismate | - |
? |
Synonyms | Comment | Organism |
---|---|---|
EntC | - |
Escherichia coli |
ICS | - |
Escherichia coli |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.05 | - |
chorismate | mutant enzyme F327Y/I346L, in 100 mM phosphate pH 7.0, 10 mM MgCl2 | Escherichia coli | |
0.067 | - |
chorismate | mutant enzyme A303T, in 100 mM phosphate pH 7.0, 10 mM MgCl2 | Escherichia coli | |
0.13 | - |
chorismate | mutant enzyme F327Y, in 100 mM phosphate pH 7.0, 10 mM MgCl2 | Escherichia coli | |
0.18 | - |
chorismate | mutant enzyme F359Q/I346L, in 100 mM phosphate pH 7.0, 10 mM MgCl2 | Escherichia coli | |
0.43 | - |
chorismate | mutant enzyme F327Y/F359Q, in 100 mM phosphate pH 7.0, 10 mM MgCl2 | Escherichia coli | |
0.55 | - |
chorismate | mutant enzyme I346L, in 100 mM phosphate pH 7.0, 10 mM MgCl2 | Escherichia coli | |
0.6 | - |
chorismate | mutant enzyme F359Q, in 100 mM phosphate pH 7.0, 10 mM MgCl2 | Escherichia coli | |
0.67 | - |
chorismate | wild type enzyme, in 100 mM phosphate pH 7.0, 10 mM MgCl2 | Escherichia coli |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.11 | - |
chorismate | mutant enzyme F327Y/I346L, in 100 mM phosphate pH 7.0, 10 mM MgCl2 | Escherichia coli | |
0.65 | - |
chorismate | mutant enzyme A303T, in 100 mM phosphate pH 7.0, 10 mM MgCl2 | Escherichia coli | |
0.69 | - |
chorismate | mutant enzyme F359Q/I346L, in 100 mM phosphate pH 7.0, 10 mM MgCl2 | Escherichia coli | |
1.71 | - |
chorismate | mutant enzyme F327Y, in 100 mM phosphate pH 7.0, 10 mM MgCl2 | Escherichia coli | |
7.24 | - |
chorismate | mutant enzyme I346L, in 100 mM phosphate pH 7.0, 10 mM MgCl2 | Escherichia coli | |
9.1 | - |
chorismate | mutant enzyme F359Q, in 100 mM phosphate pH 7.0, 10 mM MgCl2 | Escherichia coli | |
10.75 | - |
chorismate | mutant enzyme F327Y/F359Q, in 100 mM phosphate pH 7.0, 10 mM MgCl2 | Escherichia coli | |
95.7 | - |
chorismate | wild type enzyme, in 100 mM phosphate pH 7.0, 10 mM MgCl2 | Escherichia coli |