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Literature summary extracted from

  • Mueller, P.; Egorova, K.; Vorgias, C.E.; Boutou, E.; Trauthwein, H.; Verseck, S.; Antranikian, G.
    Cloning, overexpression, and characterization of a thermoactive nitrilase from the hyperthermophilic archaeon Pyrococcus abyssi (2006), Protein Expr. Purif., 47, 672-681.
    View publication on PubMed

Activating Compound

EC Number Activating Compound Comment Organism Structure
3.5.5.7 2-mercaptoethanol activates at 1 mM Pyrococcus abyssi
3.5.5.7 dithiothreitol activates at 1 mM Pyrococcus abyssi
3.5.5.7 dithiothreitol 1 mM, 23% inhibition Pyrococcus abyssi
3.5.5.7 ethylene glycol 20% increase of activity at 10% ethylene glycol Pyrococcus abyssi
3.5.5.7 L-cysteine activates at 1 mM Pyrococcus abyssi

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.5.5.7 expressed in Escherichia coli Pyrococcus abyssi
3.5.5.7 expression in Escherichia coli Pyrococcus abyssi

Inhibitors

EC Number Inhibitors Comment Organism Structure
3.5.5.7 2-mercaptoethanol 5 mM, 29% inhibition Pyrococcus abyssi
3.5.5.7 5,5'-dithiobis(2-nitrobenzoate) complete inhibition at 5 mM Pyrococcus abyssi
3.5.5.7 acetone 10%, complete inhibition; complete inhibition at 10% acetone Pyrococcus abyssi
3.5.5.7 Ag+ 1 mM, complete inhibition; complete inhibition at 1 mM Pyrococcus abyssi
3.5.5.7 Ca2+ 1 mM, 20% inhibition; 75% residual activity at 5 mM Pyrococcus abyssi
3.5.5.7 Cr3+ 5 mM, 40% inhibition; 60% residual activity at 5 mM Pyrococcus abyssi
3.5.5.7 dithiothreitol 5 mM, 50% inhibition Pyrococcus abyssi
3.5.5.7 DTNB 5 mM, complete inhibition Pyrococcus abyssi
3.5.5.7 ethanol 57% inhibition at 30%, complete inhibition at 50%; complete inhibition at 50% ethanol Pyrococcus abyssi
3.5.5.7 ethylene glycol 80% inhibition at 50%; 80% inhibition at 50% ethylene glycol Pyrococcus abyssi
3.5.5.7 Fe2+ 40% residual activity at 5 mM; 5 mM, 60% inhibition Pyrococcus abyssi
3.5.5.7 Hg2+ 1 mM, complete inhibition; complete inhibition at 1 mM Pyrococcus abyssi
3.5.5.7 iodacetamide complete inhibition at 5 mM Pyrococcus abyssi
3.5.5.7 iodacetate complete inhibition at 5 mM Pyrococcus abyssi
3.5.5.7 iodoacetamide 5 mM, complete inhibition Pyrococcus abyssi
3.5.5.7 iodoacetate 5 mM, complete inhibition Pyrococcus abyssi
3.5.5.7 L-Cys 5 mM, 27% inhibition Pyrococcus abyssi
3.5.5.7 Malononitrile concentrations above 12 mM have a strong inhibitory effect; strong inhibition at 12 mM Pyrococcus abyssi
3.5.5.7 methanol complete inhibition at 30%; complete inhibition at 30% methanol Pyrococcus abyssi
3.5.5.7 additional information EDTA and NaN3 have no influence on activity Pyrococcus abyssi
3.5.5.7 p-chloromercuribenzoate complete inhibition at 1 mM Pyrococcus abyssi
3.5.5.7 p-hydroxymercuribenzoate 1 mM, complete inhibition; complete inhibition at 1 mM Pyrococcus abyssi
3.5.5.7 PCMB 1 mM, complete inhibition Pyrococcus abyssi
3.5.5.7 phenyl hydrazine 53% residual activity at 5 mM; 5 mM, 47% inhibition Pyrococcus abyssi

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.5.5.7 3.47
-
 Malononitrile pH 7.4, 80°C Pyrococcus abyssi
3.5.5.7 3.47
-
 Malononitrile in 20 mM potassium phosphate, pH 7.4, with 1 mM dithiothreitol, at 80°C Pyrococcus abyssi
3.5.5.7 9.48
-
 Fumaronitrile pH 7.4, 80°C Pyrococcus abyssi
3.5.5.7 9.48
-
 Fumaronitrile in 20 mM potassium phosphate, pH 7.4, with 1 mM dithiothreitol, at 80°C Pyrococcus abyssi

Localization

EC Number Localization Comment Organism GeneOntology No. Textmining
3.5.5.7 intracellular
-
 Pyrococcus abyssi 5622
-

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
3.5.5.7 29800
-
 2 * 29800, gel filtration Pyrococcus abyssi
3.5.5.7 30000
-
 2 * 30000, SDS-PAGE Pyrococcus abyssi
3.5.5.7 59800
-
 gel filtration Pyrococcus abyssi
3.5.5.7 60000
-
 gel filtration Pyrococcus abyssi
3.5.5.7 60000
-
 SDS-PAGE Pyrococcus abyssi

Organism

EC Number Organism UniProt Comment Textmining
3.5.5.7 no activity in Aeropyrum pernix
-
-
-
3.5.5.7 no activity in Pyrococcus furiosus
-
-
-
3.5.5.7 no activity in Pyrococcus horikoshii
-
-
-
3.5.5.7 Pyrococcus abyssi
-
-
-
3.5.5.7 Pyrococcus abyssi GE5 / CNCM I-1302 / DSM 25543
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.5.5.7
-
 Pyrococcus abyssi
3.5.5.7 Q-Sepharose Fast Flow column chromatography and Sephadex PD-10 column gel filtration Pyrococcus abyssi

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
3.5.5.7 0.00595
-
 recombinant enzyme from crude extract Pyrococcus abyssi
3.5.5.7 0.1419
-
 recombinant enzyme after 23.85fold purification Pyrococcus abyssi
3.5.5.7 141.9
-
-
 Pyrococcus abyssi

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.5.5.7 fumaronitrile + H2O
-
 Pyrococcus abyssi fumaric acid + NH3
-
 ?
3.5.5.7 fumaronitrile + H2O preferred substrate Pyrococcus abyssi fumaric acid + NH3
-
 ?
3.5.5.7 fumaronitrile + H2O
-
 Pyrococcus abyssi GE5 / CNCM I-1302 / DSM 25543 fumaric acid + NH3
-
 ?
3.5.5.7 fumaronitrile + H2O preferred substrate Pyrococcus abyssi GE5 / CNCM I-1302 / DSM 25543 fumaric acid + NH3
-
 ?
3.5.5.7 malononitrile + H2O
-
 Pyrococcus abyssi malonic acid + NH3
-
 ?
3.5.5.7 malononitrile + H2O preferred substrate Pyrococcus abyssi malonic acid + NH3
-
 ?
3.5.5.7 malononitrile + H2O
-
 Pyrococcus abyssi GE5 / CNCM I-1302 / DSM 25543 malonic acid + NH3
-
 ?
3.5.5.7 malononitrile + H2O preferred substrate Pyrococcus abyssi GE5 / CNCM I-1302 / DSM 25543 malonic acid + NH3
-
 ?
3.5.5.7 additional information compounds with a nitrile group bound to an aromatic ring or amino acid are not hydrolyzed by recombinant nitrilase Pyrococcus abyssi ?
-
 ?
3.5.5.7 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

EC Number Subunits Comment Organism
3.5.5.7 dimer 2 * 30000, SDS-PAGE Pyrococcus abyssi
3.5.5.7 homodimer 2 * 30000, SDS-PAGE Pyrococcus abyssi
3.5.5.7 homodimer 2 * 29800, gel filtration Pyrococcus abyssi

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
3.5.5.7 80
-
-
 Pyrococcus abyssi

Temperature Range [°C]

EC Number Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
3.5.5.7 60 90
-
 Pyrococcus abyssi

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
3.5.5.7 70
-
 25 h, recombinant enzyme loses about 50% of maximal activity Pyrococcus abyssi
3.5.5.7 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
3.5.5.7 80
-
 half-life: 9 h Pyrococcus abyssi
3.5.5.7 90
-
 half-life: 6 h Pyrococcus abyssi
3.5.5.7 113
-
 Tm-value: 112.7°C Pyrococcus abyssi
3.5.5.7 120
-
 irreversible denaturation after heating to 120°C Pyrococcus abyssi

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
3.5.5.7 7.4
-
-
 Pyrococcus abyssi

pH Range

EC Number pH Minimum pH Maximum Comment Organism
3.5.5.7 6 8
-
 Pyrococcus abyssi

pI Value

EC Number Organism Comment pI Value Maximum pI Value
3.5.5.7 Pyrococcus abyssi isoelectric focusing
-
 5.3