Cloned (Comment) | Organism |
---|---|
gene pchA, overexpression of His-tagged enzyme in Escherichia coli strain BL21 (DE3) pLysS | Pseudomonas aeruginosa |
Protein Variants | Comment | Organism |
---|---|---|
E269A | site-directed mutagenesis, the residue acts as general acid, inactive mutant | Pseudomonas aeruginosa |
E313A | site-directed mutagenesis, the residue chelates Mg2+ and acts as general base, mutant with highly reduced activity compared to wild-type | Pseudomonas aeruginosa |
E313Q | site-directed mutagenesis, the residue chelates Mg2+ and acts as general base, mutant with highly reduced activity compared to wild-type | Pseudomonas aeruginosa |
K221A | site-directed mutagenesis, the residue acts as general base, mutant with highly reduced activity compared to wild-type | Pseudomonas aeruginosa |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | steady state kinetic analysis | Pseudomonas aeruginosa | |
0.0009 | - |
chorismate | pH 7.5, 25°C, wild-type enzyme | Pseudomonas aeruginosa | |
0.012 | - |
chorismate | pH 7.5, 25°C, mutant E313Q | Pseudomonas aeruginosa | |
0.017 | - |
chorismate | pH 7.5, 25°C, mutant E313A | Pseudomonas aeruginosa | |
0.021 | - |
chorismate | pH 7.5, 25°C, mutant K221A | Pseudomonas aeruginosa |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | required | Pseudomonas aeruginosa |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
Chorismate | Pseudomonas aeruginosa | - |
Isochorismate | - |
r |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Pseudomonas aeruginosa | - |
gene pchA | - |
Purification (Comment) | Organism |
---|---|
recombinant His-tagged enzyme from Escherichia coli strain BL21 (DE3) pLysS by nickel affinity chromatography, gel filtration, and ultrafitration | Pseudomonas aeruginosa |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
chorismate = isochorismate | general acid-general base catalysis with a lysine as the base and a glutamic acid as the acid, in reverse protonation states and with a conformational or binding step as the slow step | Pseudomonas aeruginosa |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
Chorismate | - |
Pseudomonas aeruginosa | Isochorismate | - |
r |
Synonyms | Comment | Organism |
---|---|---|
PchA | - |
Pseudomonas aeruginosa |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
25 | - |
assay at | Pseudomonas aeruginosa |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0455 | - |
chorismate | pH 7.5, 25°C, mutant K221A | Pseudomonas aeruginosa | |
0.108 | - |
chorismate | pH 7.5, 25°C, mutant E313A | Pseudomonas aeruginosa | |
0.75 | - |
chorismate | pH 7.5, 25°C, mutant E313Q | Pseudomonas aeruginosa | |
50.4 | - |
chorismate | pH 7.5, 25°C, wild-type enzyme | Pseudomonas aeruginosa |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.5 | - |
assay at | Pseudomonas aeruginosa |
pH Minimum | pH Maximum | Comment | Organism |
---|---|---|---|
additional information | - |
PchA pH rate profile, overview | Pseudomonas aeruginosa |
General Information | Comment | Organism |
---|---|---|
evolution | PchA is a member of the MST, i.e. menaquinone, siderophore and tryptophan, family of enzymes | Pseudomonas aeruginosa |
additional information | solvent kinetic isotope effects, overview | Pseudomonas aeruginosa |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0022 | - |
chorismate | pH 7.5, 25°C, mutant K221A | Pseudomonas aeruginosa | |
0.0064 | - |
chorismate | pH 7.5, 25°C, mutant E313A | Pseudomonas aeruginosa | |
0.06 | - |
chorismate | pH 7.5, 25°C, mutant E313Q | Pseudomonas aeruginosa | |
57 | - |
chorismate | pH 7.5, 25°C, wild-type enzyme | Pseudomonas aeruginosa |