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Literature summary for 3.5.1.B15 extracted from
- Biochemical characterization and computational identification of Mycobacterium tuberculosis pyrazinamidase in some pyrazinamide-resistant isolates of Iran (2015), Protein J., 34, 181-192 .
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| gene pncA, recombinant expression of His-tagged wild-type and mutant enzymes in Escherichia coli strain BL21(DE3), subcloning in Escherichia coli strain DH5alpha | Mycobacterium tuberculosis |
Protein Variants
| Protein Variants | Comment | Organism |
|---|---|---|
| A143T | site-directed mutagenesis, the mutation decreases the Km and kcat values of the enzyme | Mycobacterium tuberculosis |
| A143T/T168A/E173K | site-directed mutagenesis, the mutation decreases the Km and kcat values of the enzyme, the mutant shows reduced thermostability compared to wild-type | Mycobacterium tuberculosis |
| L151S | site-directed mutagenesis, the mutant has a weakened binding affinity for pyrazinamide and reduced thermostability compared to the wild-type | Mycobacterium tuberculosis |
| additional information | molecular modeling and substrate docking reveals that the wild-type has much stronger binding affinity to PZA than the mutants whereas mutant L151S has the weakest binding affinity. Molecular dynamics simulations and the essential dynamics results illustrate that the positions of the 51st to 71st residues are more dynamics in mutant L151S as compared to the other atoms in PZase. Kinetics of activation and half-life of wild-type and mutant PZases | Mycobacterium tuberculosis |
General Stability
| General Stability | Organism |
|---|---|
| kinetics of activation and half-life of wild-type and mutant PZases | Mycobacterium tuberculosis |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.431 | - |
Pyrazinamide | pH 6.5, 37°C, recombinant His-tagged mutant L151S | Mycobacterium tuberculosis |  |
| 0.46 | - |
Pyrazinamide | pH 6.5, 37°C, recombinant His-tagged mutant A143T | Mycobacterium tuberculosis | |
| 0.68 | - |
Pyrazinamide | pH 6.5, 37°C, recombinant His-tagged mutant A143T/T168A/E173K | Mycobacterium tuberculosis | |
| 0.79 | - |
Pyrazinamide | pH 6.5, 37°C, recombinant His-tagged wild-type enzyme | Mycobacterium tuberculosis | |
Metals/Ions
| Metals/Ions | Comment | Organism | Structure |
|---|---|---|---|
| Fe2+ | the iron binding site residues are Asp49, His51, His57, and His71 | Mycobacterium tuberculosis | |
Natural Substrates/ Products (Substrates)
| Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| pyrazinamide + H2O | Mycobacterium tuberculosis | - |
pyrazinoic acid + NH3 | - |
? | |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Mycobacterium tuberculosis | - |
pyrazinamide-resistant isolates from Iran | - |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| recombinant His-tagged wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) by nickel affinity chromatography | Mycobacterium tuberculosis |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| pyrazinamide + H2O | - |
Mycobacterium tuberculosis | pyrazinoic acid + NH3 | - |
? | |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| PncA | - |
Mycobacterium tuberculosis |
| PZAse | - |
Mycobacterium tuberculosis |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 37 | - |
assay at | Mycobacterium tuberculosis |
Turnover Number [1/s]
| Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.725 | - |
Pyrazinamide | pH 6.5, 37°C, recombinant His-tagged mutant L151S | Mycobacterium tuberculosis | |
| 1.08 | - |
Pyrazinamide | pH 6.5, 37°C, recombinant His-tagged mutant A143T | Mycobacterium tuberculosis | |
| 1.67 | - |
Pyrazinamide | pH 6.5, 37°C, recombinant His-tagged mutant A143T/T168A/E173K | Mycobacterium tuberculosis | |
| 2 | - |
Pyrazinamide | pH 6.5, 37°C, recombinant His-tagged wild-type enzyme | Mycobacterium tuberculosis | |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 6.5 | - |
assay at | Mycobacterium tuberculosis |
General Information
| General Information | Comment | Organism |
|---|---|---|
| additional information | the active site residues are Asp8, Lys96, and Cys138, structure-function relationship and catalytic mechanism, overview. Homology structure modeling of wild-type and mutant enzymes, molecular substrate docking and molecular dynamics simulations, overview | Mycobacterium tuberculosis |
kcat/KM [mM/s]
| kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 1.68 | - |
Pyrazinamide | pH 6.5, 37°C, recombinant His-tagged mutant L151S | Mycobacterium tuberculosis | |
| 2.35 | - |
Pyrazinamide | pH 6.5, 37°C, recombinant His-tagged mutant A143T | Mycobacterium tuberculosis | |
| 2.46 | - |
Pyrazinamide | pH 6.5, 37°C, recombinant His-tagged mutant A143T/T168A/E173K | Mycobacterium tuberculosis | |
| 2.53 | - |
Pyrazinamide | pH 6.5, 37°C, recombinant His-tagged wild-type enzyme | Mycobacterium tuberculosis | |
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