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Literature summary for 3.5.5.5 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
Search for more literature for 3.5.5.5

Application

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

Cloned(Commentary)

Cloned (Comment) Organism
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

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

Inhibitors

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

KM Value [mM]

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

Metals/Ions

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

Molecular Weight [Da]

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

Natural Substrates/ Products (Substrates)

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

Organism

Organism UniProt Comment Textmining
Alcaligenes faecalis
-
-
-
Alcaligenes faecalis ZJUTB10
-
-
-

Purification (Commentary)

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

Reaction

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

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
(R,S)-mandelonitrile + 2 H2O
-
 Alcaligenes faecalis (R)-mandelic acid + NH3 the enantiomeric excess against (R)-mandelic acid is over 99% ?
(R,S)-mandelonitrile + 2 H2O
-
 Alcaligenes faecalis ZJUTB10 (R)-mandelic acid + NH3 the enantiomeric excess against (R)-mandelic acid is over 99% ?
2-methoxyphenylacetonitrile + 2 H2O
-
 Alcaligenes faecalis 2-methoxyphenylacetic acid + NH3
-
 ?
2-methoxyphenylacetonitrile + 2 H2O
-
 Alcaligenes faecalis ZJUTB10 2-methoxyphenylacetic acid + NH3
-
 ?
3-methoxyphenylacetonitrile + 2 H2O
-
 Alcaligenes faecalis 3-methoxyphenylacetic acid + NH3
-
 ?
4-methoxyphenylacetonitrile + 2 H2O
-
 Alcaligenes faecalis 4-methoxyphenylacetic acid + NH3
-
 ?
mandelonitrile + 2 H2O
-
 Alcaligenes faecalis mandelate + NH3
-
 ?
mandelonitrile + 2 H2O
-
 Alcaligenes faecalis ZJUTB10 mandelate + NH3
-
 ?
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 ?
-
 ?
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 ?
-
 ?
phenylacetonitrile + 2 H2O
-
 Alcaligenes faecalis phenylacetic acid + NH3
-
 ?
phenylacetonitrile + 2 H2O
-
 Alcaligenes faecalis ZJUTB10 phenylacetic acid + NH3
-
 ?

Subunits

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

Temperature Optimum [°C]

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

Temperature Range [°C]

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

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
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

pH Optimum Minimum pH Optimum Maximum Comment Organism
7 8
-
 Alcaligenes faecalis

General Information

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