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Literature summary extracted from
- Lysine221 is the general base residue of the isochorismate synthase from Pseudomonas aeruginosa (PchA) in a reaction that is diffusion limited (2013), Arch. Biochem. Biophys., 538, 49-56.
Cloned(Commentary)
| EC Number | Cloned (Comment) | Organism |
|---|---|---|
| 5.4.4.2 | gene pchA, overexpression of His-tagged enzyme in Escherichia coli strain BL21 (DE3) pLysS | Pseudomonas aeruginosa |
Protein Variants
| EC Number | Protein Variants | Comment | Organism |
|---|---|---|---|
| 5.4.4.2 | E269A | site-directed mutagenesis, the residue acts as general acid, inactive mutant | Pseudomonas aeruginosa |
| 5.4.4.2 | E313A | site-directed mutagenesis, the residue chelates Mg2+ and acts as general base, mutant with highly reduced activity compared to wild-type | Pseudomonas aeruginosa |
| 5.4.4.2 | E313Q | site-directed mutagenesis, the residue chelates Mg2+ and acts as general base, mutant with highly reduced activity compared to wild-type | Pseudomonas aeruginosa |
| 5.4.4.2 | K221A | site-directed mutagenesis, the residue acts as general base, mutant with highly reduced activity compared to wild-type | Pseudomonas aeruginosa |
KM Value [mM]
| EC Number | KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|---|
| 5.4.4.2 | additional information | - |
additional information | steady state kinetic analysis | Pseudomonas aeruginosa | |
| 5.4.4.2 | 0.0009 | - |
chorismate | pH 7.5, 25°C, wild-type enzyme | Pseudomonas aeruginosa |  |
| 5.4.4.2 | 0.012 | - |
chorismate | pH 7.5, 25°C, mutant E313Q | Pseudomonas aeruginosa | |
| 5.4.4.2 | 0.017 | - |
chorismate | pH 7.5, 25°C, mutant E313A | Pseudomonas aeruginosa | |
| 5.4.4.2 | 0.021 | - |
chorismate | pH 7.5, 25°C, mutant K221A | Pseudomonas aeruginosa | |
Metals/Ions
| EC Number | Metals/Ions | Comment | Organism | Structure |
|---|---|---|---|---|
| 5.4.4.2 | Mg2+ | required | Pseudomonas aeruginosa | |
Natural Substrates/ Products (Substrates)
| EC Number | Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|---|
| 5.4.4.2 | Chorismate | Pseudomonas aeruginosa | - |
Isochorismate | - |
r | |
Organism
| EC Number | Organism | UniProt | Comment | Textmining |
|---|---|---|---|---|
| 5.4.4.2 | Pseudomonas aeruginosa | - |
gene pchA | - |
Purification (Commentary)
| EC Number | Purification (Comment) | Organism |
|---|---|---|
| 5.4.4.2 | recombinant His-tagged enzyme from Escherichia coli strain BL21 (DE3) pLysS by nickel affinity chromatography, gel filtration, and ultrafitration | Pseudomonas aeruginosa |
Reaction
| EC Number | Reaction | Comment | Organism | Reaction ID |
|---|---|---|---|---|
| 5.4.4.2 | 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 and Products (Substrate)
| EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|---|
| 5.4.4.2 | Chorismate | - |
Pseudomonas aeruginosa | Isochorismate | - |
r | |
Synonyms
| EC Number | Synonyms | Comment | Organism |
|---|---|---|---|
| 5.4.4.2 | PchA | - |
Pseudomonas aeruginosa |
Temperature Optimum [°C]
| EC Number | Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|---|
| 5.4.4.2 | 25 | - |
assay at | Pseudomonas aeruginosa |
Turnover Number [1/s]
| EC Number | Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|---|
| 5.4.4.2 | 0.0455 | - |
chorismate | pH 7.5, 25°C, mutant K221A | Pseudomonas aeruginosa | |
| 5.4.4.2 | 0.108 | - |
chorismate | pH 7.5, 25°C, mutant E313A | Pseudomonas aeruginosa | |
| 5.4.4.2 | 0.75 | - |
chorismate | pH 7.5, 25°C, mutant E313Q | Pseudomonas aeruginosa | |
| 5.4.4.2 | 50.4 | - |
chorismate | pH 7.5, 25°C, wild-type enzyme | Pseudomonas aeruginosa | |
pH Optimum
| EC Number | pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|---|
| 5.4.4.2 | 7.5 | - |
assay at | Pseudomonas aeruginosa |
pH Range
| EC Number | pH Minimum | pH Maximum | Comment | Organism |
|---|---|---|---|---|
| 5.4.4.2 | additional information | - |
PchA pH rate profile, overview | Pseudomonas aeruginosa |
General Information
| EC Number | General Information | Comment | Organism |
|---|---|---|---|
| 5.4.4.2 | evolution | PchA is a member of the MST, i.e. menaquinone, siderophore and tryptophan, family of enzymes | Pseudomonas aeruginosa |
| 5.4.4.2 | additional information | solvent kinetic isotope effects, overview | Pseudomonas aeruginosa |
kcat/KM [mM/s]
| EC Number | kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|---|
| 5.4.4.2 | 0.0022 | - |
chorismate | pH 7.5, 25°C, mutant K221A | Pseudomonas aeruginosa | |
| 5.4.4.2 | 0.0064 | - |
chorismate | pH 7.5, 25°C, mutant E313A | Pseudomonas aeruginosa | |
| 5.4.4.2 | 0.06 | - |
chorismate | pH 7.5, 25°C, mutant E313Q | Pseudomonas aeruginosa | |
| 5.4.4.2 | 57 | - |
chorismate | pH 7.5, 25°C, wild-type enzyme | Pseudomonas aeruginosa | |
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