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Literature summary extracted from

  • Liu, Z.Q.; Dong, L.Z.; Cheng, F.; Xue, Y.P.; Wang, Y.S.; Ding, J.N.; Zheng, Y.G.; Shen, Y.C.
    Gene cloning, expression, and characterization of a nitrilase from Alcaligenes faecalis ZJUTB10 (2011), J. Agric. Food Chem., 59, 11560-11570.
    View publication on PubMed

Application

EC Number Application Comment Organism
3.5.5.5 synthesis nitrilases are important industrial enzymes that convert nitriles directly into the corresponding carboxylic acids Alcaligenes faecalis

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.5.5.5 DNA and amino acid sequence determination and analysis, sequence comparison, cloning and expression as His-tagged protein in Escherichia coli strains JM109 and BL21(DE3) Alcaligenes faecalis

Protein Variants

EC Number Protein Variants Comment Organism
3.5.5.5 C163G site-directed mutagenesis Alcaligenes faecalis
3.5.5.5 E47Q site-directed mutagenesis Alcaligenes faecalis
3.5.5.5 K129I site-directed mutagenesis Alcaligenes faecalis

Inhibitors

EC Number Inhibitors Comment Organism Structure
3.5.5.5 Ag+ 94% inhibition at 5 mM Alcaligenes faecalis
3.5.5.5 Ba2+ 8% inhibition at 5 mM Alcaligenes faecalis
3.5.5.5 Co2+ 80% inhibition at 5 mM Alcaligenes faecalis
3.5.5.5 Cu2+ 84% inhibition at 5 mM Alcaligenes faecalis
3.5.5.5 Fe2+ 17% inhibition at 5 mM Alcaligenes faecalis
3.5.5.5 Fe3+ 19% inhibition at 5 mM Alcaligenes faecalis
3.5.5.5 Hg2+ complete inhibition at 5 mM Alcaligenes faecalis
3.5.5.5 Li+ 11% inhibition at 5 mM Alcaligenes faecalis
3.5.5.5 Mn2+ 22% inhibition at 5 mM Alcaligenes faecalis
3.5.5.5 Ni2+ 83% inhibition at 5 mM Alcaligenes faecalis
3.5.5.5 Zn2+ 36% inhibition at 5 mM Alcaligenes faecalis

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.5.5.5 4.74
-
 mandelonitrile pH 7.5, 40°C Alcaligenes faecalis

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
3.5.5.5 Mg2+ stimulates hydrolytic activity to 122% Alcaligenes faecalis
3.5.5.5 additional information no effects by Ca2+ and EDTA at 5 mM Alcaligenes faecalis

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
3.5.5.5 44200
-
 x * 44200, recombinant enzyme, SDS-PAGE Alcaligenes faecalis

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
3.5.5.5 mandelonitrile + 2 H2O Alcaligenes faecalis
-
 mandelate + NH3
-
 ?
3.5.5.5 mandelonitrile + 2 H2O Alcaligenes faecalis ZJUTB10
-
 mandelate + NH3
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
3.5.5.5 Alcaligenes faecalis
-
-
-
3.5.5.5 Alcaligenes faecalis ZJUTB10
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.5.5.5 recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography Alcaligenes faecalis

Reaction

EC Number Reaction Comment Organism Reaction ID
3.5.5.5 4-chlorophenylacetonitrile + 2 H2O = 4-chlorophenylacetate + NH3 reaction mechanism, overview Alcaligenes faecalis
a

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.5.5.5 (R,S)-mandelonitrile + 2 H2O
-
 Alcaligenes faecalis (R)-mandelic acid + NH3 the enantiomeric excess against (R)-mandelic acid is over 99% ?
3.5.5.5 (R,S)-mandelonitrile + 2 H2O
-
 Alcaligenes faecalis ZJUTB10 (R)-mandelic acid + NH3 the enantiomeric excess against (R)-mandelic acid is over 99% ?
3.5.5.5 2-methoxyphenylacetonitrile + 2 H2O
-
 Alcaligenes faecalis 2-methoxyphenylacetic acid + NH3
-
 ?
3.5.5.5 2-methoxyphenylacetonitrile + 2 H2O
-
 Alcaligenes faecalis ZJUTB10 2-methoxyphenylacetic acid + NH3
-
 ?
3.5.5.5 3-methoxyphenylacetonitrile + 2 H2O
-
 Alcaligenes faecalis 3-methoxyphenylacetic acid + NH3
-
 ?
3.5.5.5 4-methoxyphenylacetonitrile + 2 H2O
-
 Alcaligenes faecalis 4-methoxyphenylacetic acid + NH3
-
 ?
3.5.5.5 mandelonitrile + 2 H2O
-
 Alcaligenes faecalis mandelate + NH3
-
 ?
3.5.5.5 mandelonitrile + 2 H2O
-
 Alcaligenes faecalis ZJUTB10 mandelate + NH3
-
 ?
3.5.5.5 additional information though the enzyme exhibits activity toward both classes of aromatic and arylacetonitriles, the nitrilase shows higher activity toward the arylacetonitriles, substrate specificity and substrate docking study, overview. No activity toward aminobutytonitrile, 2,2-dimethylcyclopropyl cyanide, and 2-amino-2,3-dimethylbutanenitrile, very low activity toward 4-aminobenzonitrile and iminodiacetonitrile, overview Alcaligenes faecalis ?
-
 ?
3.5.5.5 additional information though the enzyme exhibits activity toward both classes of aromatic and arylacetonitriles, the nitrilase shows higher activity toward the arylacetonitriles, substrate specificity and substrate docking study, overview. No activity toward aminobutytonitrile, 2,2-dimethylcyclopropyl cyanide, and 2-amino-2,3-dimethylbutanenitrile, very low activity toward 4-aminobenzonitrile and iminodiacetonitrile, overview Alcaligenes faecalis ZJUTB10 ?
-
 ?
3.5.5.5 phenylacetonitrile + 2 H2O
-
 Alcaligenes faecalis phenylacetic acid + NH3
-
 ?
3.5.5.5 phenylacetonitrile + 2 H2O
-
 Alcaligenes faecalis ZJUTB10 phenylacetic acid + NH3
-
 ?

Subunits

EC Number Subunits Comment Organism
3.5.5.5 ? x * 44200, recombinant enzyme, SDS-PAGE Alcaligenes faecalis
3.5.5.5 More homology modeling and docking, three-dimensional homology model of nitrilase, overview Alcaligenes faecalis

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
3.5.5.5 40
-
-
 Alcaligenes faecalis

Temperature Range [°C]

EC Number Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
3.5.5.5 25 65 activity range, inactive above Alcaligenes faecalis

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
3.5.5.5 35 65 purified recombinant enzyme, labile at higher temperature, half-life of 57 min at 40°C, 42 min at 45°C, and 20 min at 55°C, respectively. The activity decreases markedly above 55°C Alcaligenes faecalis

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
3.5.5.5 7 8
-
 Alcaligenes faecalis

General Information

EC Number General Information Comment Organism
3.5.5.5 evolution strain ZJUTB10 nitrilase belongs to the family of arylacetonitrilases Alcaligenes faecalis