Any feedback?
Literature summary extracted from
- Enhancement of thermo-stability and product tolerance of Pseudomonas putida nitrile hydratase by fusing with self-assembling peptide (2014), J. Biosci. Bioeng., 118, 249-252.
Cloned(Commentary)
| EC Number | Cloned (Comment) | Organism |
|---|---|---|
| 4.2.1.84 | expression of His-tagged wild-type enzyme and peptide fused enzyme variants in Escherichia coli strain BL21(DE3) | Pseudomonas putida |
Protein Variants
| EC Number | Protein Variants | Comment | Organism |
|---|---|---|---|
| 4.2.1.84 | additional information | the thermostability and product tolerance of nitrile hydratase are enhanced by fusing with two of the self-assembling amphipathic peptides, EAK16 (AEAEAKAKAEAEAKAK) at the N- and C-terminus and ELK16 (LELELKLKLELELKLK) at the N-terminus. When self-assembling amphipathic peptide ELK16 is fused to the N-terminus of the enzymes beta-subunit, the resultant enzyme (SAP-NHase-2) becomes an active inclusion body, while EAK16 does not affect the enzyme solubility when fused to the enzyme's C-terminus (SAP-NHase-10) or N-termninus (AP-NHase-1) of the beta-subunit | Pseudomonas putida |
Inhibitors
| EC Number | Inhibitors | Comment | Organism | Structure |
|---|---|---|---|---|
| 4.2.1.84 | Acrylamide | NHases exhibit low product acrylamide tolerance | Pseudomonas putida |  |
Organism
| EC Number | Organism | UniProt | Comment | Textmining |
|---|---|---|---|---|
| 4.2.1.84 | Pseudomonas putida | - |
- |
- |
| 4.2.1.84 | Pseudomonas putida NRRL-18668 | - |
- |
- |
Purification (Commentary)
| EC Number | Purification (Comment) | Organism |
|---|---|---|
| 4.2.1.84 | recombinant His-tagged wild-type enzyme and peptide fused enzyme variants from Escherichia coli strain BL21(DE3) by nickel affinity chromatography and gel filtration | Pseudomonas putida |
Specific Activity [micromol/min/mg]
| EC Number | Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
|---|---|---|---|---|
| 4.2.1.84 | 13 | - |
cell extract supernatant containing recombinant His-tagged ELK-16 N-terminal fusion enzyme variant, pH 7.5, 20°C | Pseudomonas putida |
| 4.2.1.84 | 109 | - |
cell extract supernatant containing recombinant His-tagged EAK-16 N-terminal fusion enzyme variant, pH 7.5, 20°C | Pseudomonas putida |
| 4.2.1.84 | 110 | - |
cell extract supernatant containing recombinant His-tagged wild-type enzyme, pH 7.5, 20°C | Pseudomonas putida |
| 4.2.1.84 | 372 | - |
purified recombinant His-tagged ELK-16 N-terminal fusion enzyme variant, pH 7.5, 20°C | Pseudomonas putida |
| 4.2.1.84 | 426 | - |
purified recombinant His-tagged EAK-16 N-terminal fusion enzyme variant, pH 7.5, 20°C | Pseudomonas putida |
| 4.2.1.84 | 429 | - |
purified recombinant His-tagged EAK-16 C-terminal fusion enzyme variant, pH 7.5, 20°C | Pseudomonas putida |
| 4.2.1.84 | 439 | - |
purified recombinant His-tagged wild-type enzyme, pH 7.5, 20°C | 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-carbamide | i.e. nicotinamide | ? | |
| 4.2.1.84 | 3-cyanopyridine + H2O | - |
Pseudomonas putida NRRL-18668 | pyridine-3-carbamide | i.e. nicotinamide | ? | |
| 4.2.1.84 | acrylonitrile + H2O | - |
Pseudomonas putida | acrylamide | - |
? | |
| 4.2.1.84 | acrylonitrile + H2O | - |
Pseudomonas putida NRRL-18668 | acrylamide | - |
? | |
Synonyms
| EC Number | Synonyms | Comment | Organism |
|---|---|---|---|
| 4.2.1.84 | NHase | - |
Pseudomonas putida |
Temperature Optimum [°C]
| EC Number | Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|---|
| 4.2.1.84 | 20 | - |
assay at | Pseudomonas putida |
Temperature Stability [°C]
| EC Number | Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
|---|---|---|---|---|
| 4.2.1.84 | 50 | - |
wild-type enzyme, 30 min, deactivation, enzyme fused to self-assembling amphipathic peptide ELK16 (LELELKLKLELELKLK) retains 30% activity, and SAP-NHase-1 N-terminally fused to self-assembling amphipathic peptide EAK16 (AEAEAKAKAEAEAKAK) retains 45% activity, C-terminally fused enzyme SAP-NHase-10 50% activity. Treatment with buffer containing 10% acrylamide increases the thermostability so that the wild-type retains 30% activity, and the fused enzymes SAP-NHase-1, SAP-NHase-2, and SAP-NHase-10 retain 52%, 42%, and 55% activity, respectively | Pseudomonas putida |
pH Optimum
| EC Number | pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|---|
| 4.2.1.84 | 7.5 | - |
assay at | Pseudomonas putida |
Use of this online version of BRENDA is free under the CC BY 4.0 license. See terms of use for full details.
Release 2023.1 (January 2023)
Legal
Curated at
Member of
