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Literature summary for 4.2.1.84 extracted from
- Improvement of stability of nitrile hydratase via protein fragment swapping (2014), Biochem. Biophys. Res. Commun., 450, 401-408.
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| genetic organization of wild-type and chimeric mutant enzymes, expression of wild-type and recombinant chimeric mutants in Escherichia coli strain JM109 | Pseudomonas putida |
| genetic organization of wild-type and chimeric mutant enzymes, expression of wild-type and recombinant chimeric mutants in Escherichia coli strain JM109 | Comamonas testosteroni |
| genetic organization of wild-type and chimeric mutant enzymes, expression of wild-type and recombinant chimeric mutants in Escherichia coli strain JM109 | Pseudonocardia thermophila |
Protein Variants
| Protein Variants | Comment | Organism |
|---|---|---|
| additional information | homologous protein fragment swapping method is used for the improvement of the stability of NHase from Pseudomonas putida NRRL-18668, site targeted amino recombination software and molecular dynamics are used for determination of the crossover sites for fragment recombination, overview. One thermophilic NHase fragment M1 to G98 from Comamonas testosteroni 5-MGAM-4D and two fragments K165-V196 and K165-D209 from Pseudonocardia thermophila JCM3095 are selected to swap the corresponding fragments of Pseudomonas putida NHase. The chimeric NHases show 1.4 to 3.5fold enhancement in thermostability, some show reduced activity and product inhibition compared to wild-type Pseudomonas putida NHase. But mutants 3AB and 3ABC show increased activity due to altered secondary structure compared to the Pseudomonas putida wild-type | Comamonas testosteroni |
| additional information | homologous protein fragment swapping method is used for the improvement of the stability of NHase from Pseudomonas putida NRRL-18668, site targeted amino recombination software and molecular dynamics are used for determination of the crossover sites for fragment recombination, overview. One thermophilic NHase fragment M1 to G98 from Comamonas testosteroni 5-MGAM-4D and two fragments K165-V196 and K165-D209 from Pseudonocardia thermophila JCM3095 are selected to swap the corresponding fragments of Pseudomonas putida NHase. The chimeric NHases show 1.4 to 3.5fold enhancement in thermostability, some show reduced activity and product inhibition compared to wild-type Pseudomonas putida NHase. But mutants 3AB and 3ABC show increased activity due to altered secondary structure compared to the Pseudomonas putida wild-type | Pseudonocardia thermophila |
| additional information | homologous protein fragment swapping method is used for the improvement of the stability of NHase from Pseudomonas putida NRRL-18668, site targeted amino recombination software and molecular dynamics are used for determination of the crossover sites for fragment recombination, overview. One thermophilic NHase fragment M1 to G98 from Comamonas testosteroni 5-MGAM-4D and two fragments K165-V196 and K165-D209 from Pseudonocardia thermophila JCM3095 are selected to swap the corresponding fragments of Pseudomonas putida NHase. The chimeric NHases show 1.4 to 3.5fold enhancement in thermostability, some show reduced activity and product inhibition compared to wild-type Pseudomonas putida NHase. But mutants 3AB and 3ABC show increased activity due to altered secondary structure compared to the wild-type | Pseudomonas putida |
Inhibitors
| Inhibitors | Comment | Organism | Structure |
|---|---|---|---|
| additional information | product inhibition at high concentrations | Pseudomonas putida |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Comamonas testosteroni | Q5XPL4 | beta subunit | - |
| Comamonas testosteroni | Q5XPL5 | alpha subunit | - |
| Comamonas testosteroni 5-MGAM-4D | Q5XPL4 | beta subunit | - |
| Comamonas testosteroni 5-MGAM-4D | Q5XPL5 | alpha subunit | - |
| Pseudomonas putida | - |
- |
- |
| Pseudomonas putida NRRL-18668 | - |
- |
- |
| Pseudonocardia thermophila | Q7SID2 | alpha-subunit | - |
| Pseudonocardia thermophila | Q7SID3 | beta-subunit | - |
| Pseudonocardia thermophila JCM 3095 | Q7SID2 | alpha-subunit | - |
| Pseudonocardia thermophila JCM 3095 | Q7SID3 | beta-subunit | - |
Specific Activity [micromol/min/mg]
| Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
|---|---|---|---|
| 439 | - |
purified recombinant wild-type enzyme, pH 7.5, 20°C | Pseudomonas putida |
| 483 | - |
purified recombinant chimeric mutant 3ABC, pH 7.5, 20°C | Pseudomonas putida |
| 614 | - |
purified recombinant chimeric mutant 3AB, pH 7.5, 20°C | Pseudomonas putida |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| 3-cyanopyridine + H2O | - |
Pseudomonas putida | pyridine-3-carbamide | - |
? |  |
| 3-cyanopyridine + H2O | - |
Comamonas testosteroni | pyridine-3-carbamide | - |
? | |
| 3-cyanopyridine + H2O | - |
Pseudonocardia thermophila | pyridine-3-carbamide | - |
? | |
| 3-cyanopyridine + H2O | - |
Comamonas testosteroni 5-MGAM-4D | pyridine-3-carbamide | - |
? | |
| 3-cyanopyridine + H2O | - |
Pseudomonas putida NRRL-18668 | pyridine-3-carbamide | - |
? | |
| 3-cyanopyridine + H2O | - |
Pseudonocardia thermophila JCM 3095 | pyridine-3-carbamide | - |
? | |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| NHase | - |
Pseudomonas putida |
| NHase | - |
Comamonas testosteroni |
| NHase | - |
Pseudonocardia thermophila |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 20 | - |
assay at | Pseudomonas putida |
| 20 | - |
assay at | Comamonas testosteroni |
| 20 | - |
assay at | Pseudonocardia thermophila |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 7.5 | - |
assay at | Pseudomonas putida |
| 7.5 | - |
assay at | Comamonas testosteroni |
| 7.5 | - |
assay at | Pseudonocardia thermophila |
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