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Literature summary for 5.4.4.2 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)
| Cloned (Comment) | Organism |
|---|---|
| gene pchA, overexpression of His-tagged enzyme in Escherichia coli strain BL21 (DE3) pLysS | Pseudomonas aeruginosa |
Protein Variants
| 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 [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
| Metals/Ions | Comment | Organism | Structure |
|---|---|---|---|
| Mg2+ | required | Pseudomonas aeruginosa | |
Natural Substrates/ Products (Substrates)
| Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| Chorismate | Pseudomonas aeruginosa | - |
Isochorismate | - |
r | |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Pseudomonas aeruginosa | - |
gene pchA | - |
Purification (Commentary)
| 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
| 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 and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| Chorismate | - |
Pseudomonas aeruginosa | Isochorismate | - |
r | |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| PchA | - |
Pseudomonas aeruginosa |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 25 | - |
assay at | Pseudomonas aeruginosa |
Turnover Number [1/s]
| 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
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 7.5 | - |
assay at | Pseudomonas aeruginosa |
pH Range
| pH Minimum | pH Maximum | Comment | Organism |
|---|---|---|---|
| additional information | - |
PchA pH rate profile, overview | Pseudomonas aeruginosa |
General Information
| 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 [mM/s]
| 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 | |
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