Literature summary for 3.5.5.7 extracted from

  • Thuku, R.; Brady, D.; Benedik, M.; Sewell, B.
    Microbial nitrilases: Versatile, spiral forming, industrial enzymes (2009), J. Appl. Microbiol., 106, 703-727.
    View publication on PubMed

Activating Compound

Activating Compound Comment Organism Structure
Benzonitrile
-
Rhodococcus rhodochrous

Application

Application Comment Organism
industry the enzyme is engineered for the commercial production of 3-hydroxyvaleric acid Acidovorax facilis
industry the high chemical specificity and frequent enantioselectivity of nitrilases makes them attractive biocatalysts for the production of fine chemicals and pharmaceutical intermediates. Nitrilases are also used in the treatment of toxic industrial effluent and cyanide remediation Rhodococcus rhodochrous

Engineering

Protein Variants Comment Organism
L201Q the mutant shows increased activity and specificity compared to the wild type enzyme Acidovorax facilis
F168V/L201N the mutant shows increased activity and specificity compared to the wild type enzyme Acidovorax facilis
T210G inactive Acidovorax facilis

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
38000
-
x * 38000 Comamonas testosteroni
60000
-
-
Pyrococcus abyssi
40000
-
10 * 40000 Synechocystis sp.
40000
-
14 * 40000 Acidovorax facilis
40000
-
x * 40000, this enzyme exists as an inactive dimer which oligomerizes to form active spirals in the presence of benzonitrile or an autolytic cleavage of the C-terminus Rhodococcus rhodochrous
41000
-
-
Pseudomonas sp.
41000
-
14 * 41000 or 14 * 43000 Acinetobacter sp.
41000
-
14 * 41000 or 16 * 41000 Rhodococcus rhodochrous
43000
-
14 * 41000 or 14 * 43000 Acinetobacter sp.
390000
-
-
Synechocystis sp.
650000
-
-
Rhodococcus rhodochrous
570000
-
-
Acidovorax facilis
580000
-
-
Acinetobacter sp.
29800
-
2 * 29800 Pyrococcus abyssi

Organism

Organism UniProt Comment Textmining
Acinetobacter sp.
-
-
-
Acidovorax facilis
-
-
-
Pseudomonas sp.
-
-
-
Pyrococcus abyssi
-
-
-
Rhodococcus rhodochrous Q02068
-
-
Rhodococcus rhodochrous Q03217
-
-
Synechocystis sp. Q55949
-
-
Comamonas testosteroni Q59329
-
-
Pseudomonas sp. S1
-
-
-
Acinetobacter sp. AK226
-
-
-
Rhodococcus rhodochrous K22 Q02068
-
-
Acidovorax facilis 72W
-
-
-
Rhodococcus rhodochrous J1 Q03217
-
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
acrylonitrile + H2O 100% activity Pseudomonas sp. acrylic acid + NH3
-
?
acrylonitrile + H2O 128% activity compared to benzonitrile Rhodococcus rhodochrous acrylic acid + NH3
-
?
acrylonitrile + H2O 144% activity compared to racemic Ibu-CN Acinetobacter sp. acrylic acid + NH3
-
?
acrylonitrile + H2O the enzyme shows higher activity with acrylonitrile as with benzonitrile Rhodococcus rhodochrous acrylic acid + NH3
-
?
acrylonitrile + H2O 100% activity Pseudomonas sp. S1 acrylic acid + NH3
-
?
acrylonitrile + H2O 144% activity compared to racemic Ibu-CN Acinetobacter sp. AK226 acrylic acid + NH3
-
?
acrylonitrile + H2O the enzyme shows higher activity with acrylonitrile as with benzonitrile Rhodococcus rhodochrous K22 acrylic acid + NH3
-
?
acrylonitrile + H2O 128% activity compared to benzonitrile Rhodococcus rhodochrous J1 acrylic acid + NH3
-
?
fumaronitrile + H2O 100% activity Acidovorax facilis fumaric acid + NH3
-
?
fumaronitrile + H2O 12000% activity compared to benzonitrile Synechocystis sp. fumaric acid + NH3
-
?
fumaronitrile + H2O 100% activity Acidovorax facilis 72W fumaric acid + NH3
-
?
adiponitrile + H2O 100% activity Comamonas testosteroni adipic acid + NH3
-
?
benzonitrile + 2 H2O
-
Rhodococcus rhodochrous benzoic acid + NH3
-
?
benzonitrile + 2 H2O 100% activity Rhodococcus rhodochrous benzoic acid + NH3
-
?
benzonitrile + 2 H2O 100% activity Synechocystis sp. benzoic acid + NH3
-
?
benzonitrile + 2 H2O 3.8% activity compared to acrylonitrile Pseudomonas sp. benzoic acid + NH3
-
?
benzonitrile + 2 H2O 4% activity compared to adiponitrile Comamonas testosteroni benzoic acid + NH3
-
?
benzonitrile + 2 H2O 4.6% activity compared to fumaronitrile Acidovorax facilis benzoic acid + NH3
-
?
benzonitrile + 2 H2O 94% activity compared to racemic Ibu-CN Acinetobacter sp. benzoic acid + NH3
-
?
benzonitrile + 2 H2O 3.8% activity compared to acrylonitrile Pseudomonas sp. S1 benzoic acid + NH3
-
?
benzonitrile + 2 H2O 94% activity compared to racemic Ibu-CN Acinetobacter sp. AK226 benzoic acid + NH3
-
?
benzonitrile + 2 H2O
-
Rhodococcus rhodochrous K22 benzoic acid + NH3
-
?
benzonitrile + 2 H2O 4.6% activity compared to fumaronitrile Acidovorax facilis 72W benzoic acid + NH3
-
?
benzonitrile + 2 H2O 100% activity Rhodococcus rhodochrous J1 benzoic acid + NH3
-
?
3-cyanopyridine + 2 H2O
-
Rhodococcus rhodochrous nicotinic acid + NH3
-
?
3-cyanopyridine + 2 H2O
-
Rhodococcus rhodochrous J1 nicotinic acid + NH3
-
?
malonitrile + H2O
-
Pyrococcus abyssi malonic acid + NH3
-
?
racemic Ibu-CN + 2 H2O 100% activity Acinetobacter sp. ibuprofen + NH3
-
?
racemic Ibu-CN + 2 H2O 100% activity Acinetobacter sp. AK226 ibuprofen + NH3
-
?

Subunits

Subunits Comment Organism
monomer 1 * 41000 Pseudomonas sp.
? x * 38000 Comamonas testosteroni
? x * 40000, this enzyme exists as an inactive dimer which oligomerizes to form active spirals in the presence of benzonitrile or an autolytic cleavage of the C-terminus Rhodococcus rhodochrous
homodimer 2 * 29800 Pyrococcus abyssi
homodecamer 10 * 40000 Synechocystis sp.
homotetradecamer 14 * 40000 Acidovorax facilis
homotetradecamer 14 * 41000 or 14 * 43000 Acinetobacter sp.
homotetradecamer or homohexacecamer 14 * 41000 or 16 * 41000 Rhodococcus rhodochrous

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
50
-
-
Acinetobacter sp.
50
-
-
Synechocystis sp.
50
-
-
Rhodococcus rhodochrous
45
-
-
Rhodococcus rhodochrous
65
-
-
Acidovorax facilis
30
-
-
Comamonas testosteroni
100
-
-
Pyrococcus abyssi

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
8
-
-
Acinetobacter sp.
7.4
-
-
Pyrococcus abyssi
7.6
-
-
Rhodococcus rhodochrous
7
-
-
Synechocystis sp.
7
-
-
Comamonas testosteroni
8 9
-
Acidovorax facilis
5.5
-
-
Rhodococcus rhodochrous