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Literature summary for 3.5.5.7 extracted from
- Cloning, overexpression, and characterization of a thermoactive nitrilase from the hyperthermophilic archaeon Pyrococcus abyssi (2006), Protein Expr. Purif., 47, 672-681.
Activating Compound
| Activating Compound | Comment | Organism | Structure |
|---|---|---|---|
| 2-mercaptoethanol | activates at 1 mM | Pyrococcus abyssi |  |
| dithiothreitol | activates at 1 mM | Pyrococcus abyssi | |
| dithiothreitol | 1 mM, 23% inhibition | Pyrococcus abyssi | |
| ethylene glycol | 20% increase of activity at 10% ethylene glycol | Pyrococcus abyssi | |
| L-cysteine | activates at 1 mM | Pyrococcus abyssi | |
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| expressed in Escherichia coli | Pyrococcus abyssi |
| expression in Escherichia coli | Pyrococcus abyssi |
Inhibitors
| Inhibitors | Comment | Organism | Structure |
|---|---|---|---|
| 2-mercaptoethanol | 5 mM, 29% inhibition | Pyrococcus abyssi | |
| 5,5'-dithiobis(2-nitrobenzoate) | complete inhibition at 5 mM | Pyrococcus abyssi | |
| acetone | 10%, complete inhibition; complete inhibition at 10% acetone | Pyrococcus abyssi | |
| Ag+ | 1 mM, complete inhibition; complete inhibition at 1 mM | Pyrococcus abyssi | |
| Ca2+ | 1 mM, 20% inhibition; 75% residual activity at 5 mM | Pyrococcus abyssi | |
| Cr3+ | 5 mM, 40% inhibition; 60% residual activity at 5 mM | Pyrococcus abyssi | |
| dithiothreitol | 5 mM, 50% inhibition | Pyrococcus abyssi | |
| DTNB | 5 mM, complete inhibition | Pyrococcus abyssi | |
| ethanol | 57% inhibition at 30%, complete inhibition at 50%; complete inhibition at 50% ethanol | Pyrococcus abyssi | |
| ethylene glycol | 80% inhibition at 50%; 80% inhibition at 50% ethylene glycol | Pyrococcus abyssi | |
| Fe2+ | 40% residual activity at 5 mM; 5 mM, 60% inhibition | Pyrococcus abyssi | |
| Hg2+ | 1 mM, complete inhibition; complete inhibition at 1 mM | Pyrococcus abyssi | |
| iodacetamide | complete inhibition at 5 mM | Pyrococcus abyssi | |
| iodacetate | complete inhibition at 5 mM | Pyrococcus abyssi | |
| iodoacetamide | 5 mM, complete inhibition | Pyrococcus abyssi | |
| iodoacetate | 5 mM, complete inhibition | Pyrococcus abyssi | |
| L-Cys | 5 mM, 27% inhibition | Pyrococcus abyssi | |
| Malononitrile | concentrations above 12 mM have a strong inhibitory effect; strong inhibition at 12 mM | Pyrococcus abyssi | |
| methanol | complete inhibition at 30%; complete inhibition at 30% methanol | Pyrococcus abyssi | |
| additional information | EDTA and NaN3 have no influence on activity | Pyrococcus abyssi | |
| p-chloromercuribenzoate | complete inhibition at 1 mM | Pyrococcus abyssi | |
| p-hydroxymercuribenzoate | 1 mM, complete inhibition; complete inhibition at 1 mM | Pyrococcus abyssi | |
| PCMB | 1 mM, complete inhibition | Pyrococcus abyssi | |
| phenyl hydrazine | 53% residual activity at 5 mM; 5 mM, 47% inhibition | Pyrococcus abyssi | |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 3.47 | - |
Malononitrile | pH 7.4, 80°C | Pyrococcus abyssi | |
| 3.47 | - |
Malononitrile | in 20 mM potassium phosphate, pH 7.4, with 1 mM dithiothreitol, at 80°C | Pyrococcus abyssi | |
| 9.48 | - |
Fumaronitrile | pH 7.4, 80°C | Pyrococcus abyssi | |
| 9.48 | - |
Fumaronitrile | in 20 mM potassium phosphate, pH 7.4, with 1 mM dithiothreitol, at 80°C | Pyrococcus abyssi | |
Localization
| Localization | Comment | Organism | GeneOntology No. | Textmining |
|---|---|---|---|---|
| intracellular | - |
Pyrococcus abyssi | 5622 | - |
Molecular Weight [Da]
| Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
|---|---|---|---|
| 29800 | - |
2 * 29800, gel filtration | Pyrococcus abyssi |
| 30000 | - |
2 * 30000, SDS-PAGE | Pyrococcus abyssi |
| 59800 | - |
gel filtration | Pyrococcus abyssi |
| 60000 | - |
gel filtration | Pyrococcus abyssi |
| 60000 | - |
SDS-PAGE | Pyrococcus abyssi |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| no activity in Aeropyrum pernix | - |
- |
- |
| no activity in Pyrococcus furiosus | - |
- |
- |
| no activity in Pyrococcus horikoshii | - |
- |
- |
| Pyrococcus abyssi | - |
- |
- |
| Pyrococcus abyssi GE5 / CNCM I-1302 / DSM 25543 | - |
- |
- |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
- |
Pyrococcus abyssi |
| Q-Sepharose Fast Flow column chromatography and Sephadex PD-10 column gel filtration | Pyrococcus abyssi |
Specific Activity [micromol/min/mg]
| Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
|---|---|---|---|
| 0.00595 | - |
recombinant enzyme from crude extract | Pyrococcus abyssi |
| 0.1419 | - |
recombinant enzyme after 23.85fold purification | Pyrococcus abyssi |
| 141.9 | - |
- |
Pyrococcus abyssi |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| fumaronitrile + H2O | - |
Pyrococcus abyssi | fumaric acid + NH3 | - |
? | |
| fumaronitrile + H2O | preferred substrate | Pyrococcus abyssi | fumaric acid + NH3 | - |
? | |
| fumaronitrile + H2O | - |
Pyrococcus abyssi GE5 / CNCM I-1302 / DSM 25543 | fumaric acid + NH3 | - |
? | |
| fumaronitrile + H2O | preferred substrate | Pyrococcus abyssi GE5 / CNCM I-1302 / DSM 25543 | fumaric acid + NH3 | - |
? | |
| malononitrile + H2O | - |
Pyrococcus abyssi | malonic acid + NH3 | - |
? | |
| malononitrile + H2O | preferred substrate | Pyrococcus abyssi | malonic acid + NH3 | - |
? | |
| malononitrile + H2O | - |
Pyrococcus abyssi GE5 / CNCM I-1302 / DSM 25543 | malonic acid + NH3 | - |
? | |
| malononitrile + H2O | preferred substrate | Pyrococcus abyssi GE5 / CNCM I-1302 / DSM 25543 | malonic acid + NH3 | - |
? | |
| additional information | compounds with a nitrile group bound to an aromatic ring or amino acid are not hydrolyzed by recombinant nitrilase | Pyrococcus abyssi | ? | - |
? | |
| additional information | compounds with a nitrile group bound to an aromatic ring or amino acid are not hydrolyzed by recombinant nitrilase | Pyrococcus abyssi GE5 / CNCM I-1302 / DSM 25543 | ? | - |
? | |
Subunits
| Subunits | Comment | Organism |
|---|---|---|
| dimer | 2 * 30000, SDS-PAGE | Pyrococcus abyssi |
| homodimer | 2 * 30000, SDS-PAGE | Pyrococcus abyssi |
| homodimer | 2 * 29800, gel filtration | Pyrococcus abyssi |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 80 | - |
- |
Pyrococcus abyssi |
Temperature Range [°C]
| Temperature Minimum [°C] | Temperature Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 60 | 90 | - |
Pyrococcus abyssi |
Temperature Stability [°C]
| Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 70 | - |
25 h, recombinant enzyme loses about 50% of maximal activity | Pyrococcus abyssi |
| 70 | 90 | the recombinant enzyme is highly thermostable with a half-life of 25 h at 70°C, 9 h at 80°C, and 6 h at 90°C | Pyrococcus abyssi |
| 80 | - |
half-life: 9 h | Pyrococcus abyssi |
| 90 | - |
half-life: 6 h | Pyrococcus abyssi |
| 113 | - |
Tm-value: 112.7°C | Pyrococcus abyssi |
| 120 | - |
irreversible denaturation after heating to 120°C | Pyrococcus abyssi |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 7.4 | - |
- |
Pyrococcus abyssi |
pH Range
| pH Minimum | pH Maximum | Comment | Organism |
|---|---|---|---|
| 6 | 8 | - |
Pyrococcus abyssi |
pI Value
| Organism | Comment | pI Value Maximum | pI Value |
|---|---|---|---|
| Pyrococcus abyssi | isoelectric focusing | - |
5.3 |
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