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Literature summary extracted from

  • Zhang, Z.J.; Xu, J.H.; He, Y.C.; Ouyang, L.M.; Liu, Y.Y.
    Cloning and biochemical properties of a highly thermostable and enantioselective nitrilase from Alcaligenes sp. ECU0401 and its potential for (R)-(-)-mandelic acid production (2011), Bioprocess Biosyst. Eng., 34, 315-322.
    View publication on PubMed

Activating Compound

EC Number Activating Compound Comment Organism Structure
3.5.5.1 EDTA slightly activating at 1 mM, not at 5 mM Alcaligenes sp.

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.5.5.1 expression of His6-tagged enzyme in Escherichia coli strain BL21(DE3)/pSH Alcaligenes sp.

Inhibitors

EC Number Inhibitors Comment Organism Structure
3.5.5.1 Ag+
-
 Alcaligenes sp.
3.5.5.1 Ca2+
-
 Alcaligenes sp.
3.5.5.1 Co2+
-
 Alcaligenes sp.
3.5.5.1 Cu2+
-
 Alcaligenes sp.
3.5.5.1 EDTA
-
 Alcaligenes sp.
3.5.5.1 Fe2+
-
 Alcaligenes sp.
3.5.5.1 Fe3+
-
 Alcaligenes sp.
3.5.5.1 Hg2+
-
 Alcaligenes sp.
3.5.5.1 Li+
-
 Alcaligenes sp.
3.5.5.1 Mg2+
-
 Alcaligenes sp.
3.5.5.1 Ni2+
-
 Alcaligenes sp.
3.5.5.1 Pb2+
-
 Alcaligenes sp.
3.5.5.1 Zn2+
-
 Alcaligenes sp.

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.5.5.1 21.8
-
 (R,S)-2-hydroxy-2-phenylacetonitrile pH 8.0, 40°C, recombinant enzyme Alcaligenes sp.

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
3.5.5.1 Co2+ slightly activating at 1 mM, slightly inhibitory at 5 mM Alcaligenes sp.
3.5.5.1 Li+ activating at 1 mM to 133%, slightly inhibitory at 5 mM Alcaligenes sp.
3.5.5.1 Mn2+ slightly activating at 1 mM, not at 5 mM Alcaligenes sp.

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
3.5.5.1 376000
-
 recombinant enzyme, gel filtration Alcaligenes sp.

Organism

EC Number Organism UniProt Comment Textmining
3.5.5.1 Alcaligenes sp. C5IIS9
-
-
3.5.5.1 Alcaligenes sp. ECU0401 C5IIS9
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.5.5.1 recombinant His6-tagged enzyme from Escherichia coli strain BL21(DE3)/pSH by nickel affinity chromatography Alcaligenes sp.

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
3.5.5.1 19
-
 substrate mandelonitrile, purified recombinant enzyme, pH 8.0, 40°C Alcaligenes sp.

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.5.5.1 (R,S)-2-hydroxy-2-phenylacetonitrile + 2 H2O i.e. (R,S)-mandelonitrile, preferred substrate, highly enantioselective reaction Alcaligenes sp. (R)-2-hydroxy-2-phenylacetic acid + NH3
-
 ?
3.5.5.1 (R,S)-2-hydroxy-2-phenylacetonitrile + 2 H2O i.e. (R,S)-mandelonitrile, preferred substrate, highly enantioselective reaction Alcaligenes sp. ECU0401 (R)-2-hydroxy-2-phenylacetic acid + NH3
-
 ?
3.5.5.1 2-chloromandelonitrile + 2 H2O
-
 Alcaligenes sp. 2-chloromandelic acid + NH3
-
 ?
3.5.5.1 2-chloromandelonitrile + 2 H2O
-
 Alcaligenes sp. ECU0401 2-chloromandelic acid + NH3
-
 ?
3.5.5.1 2-phenylglycinonitrile + 2 H2O
-
 Alcaligenes sp. amino(phenyl)acetic acid + NH3
-
 ?
3.5.5.1 acetonitrile + 2 H2O
-
 Alcaligenes sp. acetic acid + NH3
-
 ?
3.5.5.1 acetonitrile + 2 H2O
-
 Alcaligenes sp. ECU0401 acetic acid + NH3
-
 ?
3.5.5.1 benzyl cyanide + 2 H2O
-
 Alcaligenes sp. phenylacetic acid + NH3
-
 ?
3.5.5.1 glycolonitrile + 2 H2O
-
 Alcaligenes sp. glycolic acid + NH3
-
 ?
3.5.5.1 iminodiacetonitrile + 2 H2O
-
 Alcaligenes sp. iminodiacetic acid + 2 NH3
-
 ?
3.5.5.1 iminodiacetonitrile + 2 H2O
-
 Alcaligenes sp. ECU0401 iminodiacetic acid + 2 NH3
-
 ?
3.5.5.1 additional information substrate specificity, overview. No activity with diaminomaleonitrile, 2,3-dicyanopyrazine, 4-hydroxybenzonitrile, 2-cyano-3-phenylpropionitrile, 4-aminophthalonitrile, and 4-nitrophthalonitrile Alcaligenes sp. ?
-
 ?
3.5.5.1 additional information substrate specificity, overview. No activity with diaminomaleonitrile, 2,3-dicyanopyrazine, 4-hydroxybenzonitrile, 2-cyano-3-phenylpropionitrile, 4-aminophthalonitrile, and 4-nitrophthalonitrile Alcaligenes sp. ECU0401 ?
-
 ?

Subunits

EC Number Subunits Comment Organism
3.5.5.1 decamer
-
 Alcaligenes sp.

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
3.5.5.1 40 45
-
 Alcaligenes sp.

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
3.5.5.1 8
-
-
 Alcaligenes sp.

pH Stability

EC Number pH Stability pH Stability Maximum Comment Organism
3.5.5.1 9
-
 purified recombinant enzyme, half-life is 4 h Alcaligenes sp.

pI Value

EC Number Organism Comment pI Value Maximum pI Value
3.5.5.1 Alcaligenes sp. two-dimensional electrophoresis
-
 5.28