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Literature summary extracted from
- Improving the thermostability and catalytic efficiency of the subunit-fused nitrile hydratase by semi-rational engineering (2018), ChemCatChem, 10, 1370-1375 .
No PubMed abstract available
Application
| EC Number | Application | Comment | Organism |
|---|---|---|---|
| 4.2.1.84 | synthesis | industrial production of highly purified acrylamide and nicotinamide. The thermostability and catalytic efficiency of the subunit-fused nitrile hydratase is improved by semi-rational engineering | Pseudomonas putida |
Cloned(Commentary)
| EC Number | Cloned (Comment) | Organism |
|---|---|---|
| 4.2.1.84 | - |
Pseudomonas putida |
Protein Variants
| EC Number | Protein Variants | Comment | Organism |
|---|---|---|---|
| 4.2.1.84 | M150C | beta-subunit mutant enzyme, 32% increase in half-life at 50°C, the kcat/Km value is 1.1fold higher than the kcat/Km value of the wild-type enzyme | Pseudomonas putida |
| 4.2.1.84 | S189E | beta-subunit mutant enzyme, 107% increase in half-life at 50°C, the kcat/Km value is 2.2fold higher than the kcat/Km value of the wild-type enzyme | Pseudomonas putida |
| 4.2.1.84 | T173Y | beta-subunit mutant enzyme, 7% increase in half-life at 50°C, the kcat/Km value is 1.5fold higher than the kcat/Km value of the wild-type enzyme | Pseudomonas putida |
KM Value [mM]
| EC Number | KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|---|
| 4.2.1.84 | 36.3 | - |
3-Cyanopyridine | 25°C, pH not specified in the publication, beta-subunit mutant enzyme S189E | Pseudomonas putida |  |
| 4.2.1.84 | 62.9 | - |
3-Cyanopyridine | 25°C, pH not specified in the publication, subunit-fused wild-type nitrile hydratase | Pseudomonas putida | |
| 4.2.1.84 | 64.9 | - |
3-Cyanopyridine | 25°C, pH not specified in the publication, beta-subunit mutant enzyme T173Y | Pseudomonas putida | |
| 4.2.1.84 | 72.8 | - |
3-Cyanopyridine | 25°C, pH not specified in the publication, beta-subunit mutant enzyme M150C | Pseudomonas putida | |
Organism
| EC Number | Organism | UniProt | Comment | Textmining |
|---|---|---|---|---|
| 4.2.1.84 | Pseudomonas putida | P97051 AND P97052 | P97051: alpha-subunit, P97052: beta-subunit | - |
| 4.2.1.84 | Pseudomonas putida NRRL-18668 | P97051 AND P97052 | P97051: alpha-subunit, P97052: beta-subunit | - |
Purification (Commentary)
| EC Number | Purification (Comment) | Organism |
|---|---|---|
| 4.2.1.84 | - |
Pseudomonas putida |
Substrates and Products (Substrate)
| EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|---|
| 4.2.1.84 | 3-cyanopyridine + H2O | - |
Pseudomonas putida | pyridine-3-carboxamide | - |
? | |
| 4.2.1.84 | 3-cyanopyridine + H2O | - |
Pseudomonas putida NRRL-18668 | pyridine-3-carboxamide | - |
? | |
Temperature Stability [°C]
| EC Number | Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
|---|---|---|---|---|
| 4.2.1.84 | 50 | - |
t1/2: 26 min (subunit-fused wild-type nitrile hydratase), 36 min (beta-subunit mutant enzyme A119D), 32 min (beta-subunit mutant enzyme V139A), 34 min (beta-subunit mutant enzyme N141G), 13 min (beta-subunit mutant enzyme T148G), 32 min (beta-subunit mutant enzyme R149G), 40 min (beta-subunit mutant enzyme M150C), 29 min (beta-subunit mutant enzyme G156M), 30 min (beta-subunit mutant enzyme G156S), 25 min (beta-subunit mutant enzyme G156Q), 24 min (beta-subunit mutant enzyme G156N), 32 min (beta-subunit mutant enzyme G156E), 23 min (beta-subunit mutant enzyme H165G), 32 min (beta-subunit mutant enzyme T173Y), 38 min (beta-subunit mutant enzyme T173F), 24 min (beta-subunit mutant enzyme V188G), above 60 min (beta-subunit mutant enzyme S189E), 28 min (beta-subunit mutant enzyme Q198G) | Pseudomonas putida |
| 4.2.1.84 | 55 | - |
Tm value of subunit-fused wild-type nitrile hydratase is 54.5°C, Tm value of beta-subunit mutant enzyme T173Y is 55°C | Pseudomonas putida |
| 4.2.1.84 | 56 | - |
Tm value of beta-subunit mutant enzyme M150C is 56.2°C | Pseudomonas putida |
| 4.2.1.84 | 58 | - |
Tm value of beta-subunit mutant enzyme S189E | Pseudomonas putida |
Turnover Number [1/s]
| EC Number | Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|---|
| 4.2.1.84 | 720.6 | - |
3-Cyanopyridine | 25°C, pH not specified in the publication, subunit-fused wild-type nitrile hydratase | Pseudomonas putida | |
| 4.2.1.84 | 922.3 | - |
3-Cyanopyridine | 25°C, pH not specified in the publication, beta-subunit mutant enzyme M150C | Pseudomonas putida | |
| 4.2.1.84 | 1112 | - |
3-Cyanopyridine | 25°C, pH not specified in the publication, beta-subunit mutant enzyme T173Y | Pseudomonas putida | |
| 4.2.1.84 | 8935.6 | - |
3-Cyanopyridine | 25°C, pH not specified in the publication, beta-subunit mutant enzyme S189E | Pseudomonas putida | |
kcat/KM [mM/s]
| EC Number | kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|---|
| 4.2.1.84 | 11.5 | - |
3-Cyanopyridine | 25°C, pH not specified in the publication, subunit-fused wild-type nitrile hydratase | Pseudomonas putida | |
| 4.2.1.84 | 12.7 | - |
3-Cyanopyridine | 25°C, pH not specified in the publication, beta-subunit mutant enzyme M150C | Pseudomonas putida | |
| 4.2.1.84 | 17.1 | - |
3-Cyanopyridine | 25°C, pH not specified in the publication, beta-subunit mutant enzyme T173Y | Pseudomonas putida | |
| 4.2.1.84 | 25.8 | - |
3-Cyanopyridine | 25°C, pH not specified in the publication, beta-subunit mutant enzyme S189E | Pseudomonas putida | |
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