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Literature summary for 3.5.5.1 extracted from

  • Liu, Z.Q.; Lu, M.M.; Zhang, X.H.; Cheng, F.; Xu, J.M.; Xue, Y.P.; Jin, L.Q.; Wang, Y.S.; Zheng, Y.G.
    Significant improvement of the nitrilase activity by semi-rational protein engineering and its application in the production of iminodiacetic acid (2018), Int. J. Biol. Macromol., 116, 563-571 .
    View publication on PubMed
Search for more literature for 3.5.5.1

Activating Compound

Activating Compound Comment Organism Structure
EDTA activates wild-type nitrilase and mutant enzyme F168V/T201N/S192F/M191T/F192S Acidovorax facilis

Application

Application Comment Organism
synthesis mutant nitrilase F168V/T201N/S192F/M191T/F192S is promising in applications for the upscale production of iminodiacetic acid Acidovorax facilis

Cloned(Commentary)

Cloned (Comment) Organism
expression in Escherichia coli BL21 (DE3) Acidovorax facilis

Protein Variants

Protein Variants Comment Organism
F168V/T201N/S192F/M191T/F192S the mutant enzyme shows 136% improvement in specific activity. Vmax and kcat are enhanced 1.23fold and 1.23fold, while the Km is decreased 1.53fold Acidovorax facilis

Inhibitors

Inhibitors Comment Organism Structure
Cu2+
-
 Acidovorax facilis
Tween 80
-
 Acidovorax facilis
Zn2+
-
 Acidovorax facilis

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
536.2
-
 iminodiacetonitrile 35°C, pH 7.5, mutant enzyme F168V/T201N/S192F/M191T/F192S Acidovorax facilis
821.5
-
 iminodiacetonitrile 35°C, pH 7.5, wild-type enzyme Acidovorax facilis

Metals/Ions

Metals/Ions Comment Organism Structure
Co2+ 5 mM, stimulates wild-type nitrilase 4.8fold and the mutant enzyme F168V/T201N/S192F/M191T/F192S 3.3fold Acidovorax facilis

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
43000
-
 wild-type enzyme and mutant enzyme F168V/T201N/S192F/M191T/F192S, SDS-PAGE Acidovorax facilis

Organism

Organism UniProt Comment Textmining
Acidovorax facilis Q1W2L4
-
-

Purification (Commentary)

Purification (Comment) Organism
-
 Acidovorax facilis

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
adiponitrile + 2 H2O
-
 Acidovorax facilis adipic acid + 2 NH3
-
 ?
iminodiacetonitrile + 2 H2O high activity Acidovorax facilis iminodiacetic acid + 2 NH3
-
 ?

Subunits

Subunits Comment Organism
? x * 43000, wild-type enzyme and mutant enzyme F168V/T201N/S192F/M191T/F192S, SDS-PAGE Acidovorax facilis

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
40
-
 mutant enzyme F168V/T201N/S192F/M191T/F192S Acidovorax facilis
45
-
 wild-type enzyme Acidovorax facilis

Temperature Range [°C]

Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
30 60 30°C: about 85% of maximal activity, 60°C: about 50% of maximal activity Acidovorax facilis

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
35
-
 half-life of mutant enzyme F168V/T201N/S192F/M191T/F192S: 17.39 h. Half-life of wild-type enzyme 14.51 h Acidovorax facilis
45
-
 wild-type nitrilase and mutant enzyme F168V/T201N/S192F/M191T/F192S are not thermally stable at temperatures higher than 45°C Acidovorax facilis

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
7.47
-
 iminodiacetonitrile 35°C, pH 7.5, wild-type enzyme Acidovorax facilis
9.19
-
 iminodiacetonitrile 35°C, pH 7.5, mutant enzyme F168V/T201N/S192F/M191T/F192S Acidovorax facilis

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7
-
 wild-type enzyme and mutant enzyme F168V/T201N/S192F/M191T/F192S Acidovorax facilis