BRENDA - Enzyme Database show

Characterization of a new cobalt-containing nitrile hydratase purified from urea-induced cells of Rhodococcus rhodochrous J1

Nagasawa, T.; Takeuchi, K.; Yamada, H.; Eur. J. Biochem. 196, 581-589 (1991)

Data extracted from this reference:

General Stability
EC Number
General Stability
Organism
4.2.1.84
organic acids stabilize, stable for more than 1 month in 0.1 M HEPES/KOH, pH 7.2, with 44 mM n-butyric acid, n-valeric acid, isovaleric acid, isobutyric acid or n-caproic acid
Rhodococcus rhodochrous
Inhibitors
EC Number
Inhibitors
Commentary
Organism
Structure
4.2.1.84
Ag+
0.01-1 mM AgNO3
Rhodococcus rhodochrous
4.2.1.84
diethyldithiocarbamate
0.1 mM
Rhodococcus rhodochrous
4.2.1.84
Hg2+
HgCl2, 0.01-1 mM
Rhodococcus rhodochrous
4.2.1.84
hydroxylamine
1 mM
Rhodococcus rhodochrous
4.2.1.84
KCN
0.01 mM
Rhodococcus rhodochrous
4.2.1.84
Na2S2O4
1 mM
Rhodococcus rhodochrous
4.2.1.84
phenylhydrazine
1 mM
Rhodococcus rhodochrous
4.2.1.84
Semicarbazide
1 mM, weak inhibition
Rhodococcus rhodochrous
KM Value [mM]
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
4.2.1.84
1.85
-
Isobutyronitrile
pH 7.0, 20ºC, H-NHase
Rhodococcus rhodochrous
4.2.1.84
1.89
-
acrylonitrile
pH 7.0, 20ºC, H-NHase
Rhodococcus rhodochrous
4.2.1.84
1.92
-
Propionitrile
pH 7.0, 20ºC, H-NHase
Rhodococcus rhodochrous
4.2.1.84
4.88
-
Crotononitrile
pH 7.0, 20ºC, H-NHase
Rhodococcus rhodochrous
4.2.1.84
6.76
-
Methacrylonitrile
pH 7.0, 20ºC
Rhodococcus rhodochrous
4.2.1.84
10
-
Cyanopyrazine
pH 7.0, 20ºC
Rhodococcus rhodochrous
4.2.1.84
12.2
-
Chloroacetonitrile
pH 7.0, 20ºC, H-NHase
Rhodococcus rhodochrous
4.2.1.84
18.5
-
4-cyanopyridine
pH 7.0, 20ºC, H-NHase
Rhodococcus rhodochrous
4.2.1.84
21.7
-
2-Cyanopyridine
pH 7.0, 20ºC, H-NHase
Rhodococcus rhodochrous
4.2.1.84
21.7
-
n-butyronitrile
pH 7.0, 20ºC, H-NHase
Rhodococcus rhodochrous
4.2.1.84
28.6
-
Benzonitrile
pH 7.0, 20ºC, H-NHase
Rhodococcus rhodochrous
4.2.1.84
200
-
3-Cyanopyridine
pH 7.0, 20ºC, H-NHase
Rhodococcus rhodochrous
Metals/Ions
EC Number
Metals/Ions
Commentary
Organism
Structure
4.2.1.84
Co2+
contains approximately 11-12 mol cobalt/mol enzyme
Rhodococcus rhodochrous
Molecular Weight [Da]
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
4.2.1.84
26000
-
x * 26000, alpha, + x * 29000, beta, SDS-PAGE
Rhodococcus rhodochrous
4.2.1.84
29000
-
x * 26000, alpha, + x * 29000, beta, SDS-PAGE
Rhodococcus rhodochrous
4.2.1.84
505000
530000
sedimentation equilibrium method, gel permeation HPLC
Rhodococcus rhodochrous
Organism
EC Number
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
4.2.1.84
Rhodococcus rhodochrous
-
2 kinds of enzyme, a high molecular mass: H-NHase, and a low molecular mass enzyme: L-NHase
-
4.2.1.84
Rhodococcus rhodochrous J1
-
2 kinds of enzyme, a high molecular mass: H-NHase, and a low molecular mass enzyme: L-NHase
-
Purification (Commentary)
EC Number
Commentary
Organism
4.2.1.84
; ammonium sulfate fractionation, chromatography on DEAE-Sephacel, phenyl-Sepharose and Sephacryl S-300, 3.31fold purification
Rhodococcus rhodochrous
Specific Activity [micromol/min/mg]
EC Number
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
4.2.1.84
additional information
-
-
Rhodococcus rhodochrous
4.2.1.84
371
-
purified enzyme
Rhodococcus rhodochrous
Storage Stability
EC Number
Storage Stability
Organism
4.2.1.84
organic acids stabilize, stable for more than 1 month in 0.1 M HEPES/KOH, pH 7.2, with 44 mM n-butyric acid, n-valeric acid, isovaleric acid, isobutyric acid or n-caproic acid
Rhodococcus rhodochrous
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
4.2.1.84
2-cyanopyridine + H2O
-
648439
Rhodococcus rhodochrous
pyridine-2-carbamide
-
-
-
?
4.2.1.84
3-cyanopyridine + H2O
-
648439
Rhodococcus rhodochrous
pyridine-3-carbamide
i.e. nicotinamide
648439
Rhodococcus rhodochrous
?
4.2.1.84
4-cyanopyridine + H2O
-
648439
Rhodococcus rhodochrous
pyridine-4-carbamide
-
-
-
?
4.2.1.84
acetonitrile + H2O
best substrate
648439
Rhodococcus rhodochrous
acetamide
-
-
-
?
4.2.1.84
acetonitrile + H2O
best substrate
648439
Rhodococcus rhodochrous J1
acetamide
-
-
-
?
4.2.1.84
acrylonitrile + H2O
-
648439
Rhodococcus rhodochrous
2-propenoic acid amide
i.e. acrylic acid amide
648439
Rhodococcus rhodochrous
?
4.2.1.84
benzonitrile + H2O
-
648439
Rhodococcus rhodochrous
benzoic acid amide
-
-
-
?
4.2.1.84
chloroacetonitrile + H2O
-
648439
Rhodococcus rhodochrous
chloroacetamide
-
-
-
?
4.2.1.84
chloroacetonitrile + H2O
-
648439
Rhodococcus rhodochrous J1
chloroacetamide
-
-
-
?
4.2.1.84
crotononitrile + H2O
19% of the activity with propionitrile
648439
Rhodococcus rhodochrous
(E)-2-butenoic acid amide
-
-
-
?
4.2.1.84
cyanopyrazine + H2O
-
648439
Rhodococcus rhodochrous
pyrazincarbamide
-
-
-
?
4.2.1.84
cyclopropylcyanide + H2O
-
648439
Rhodococcus rhodochrous
?
-
-
-
?
4.2.1.84
ethylene cyanhydrine + H2O
-
648439
Rhodococcus rhodochrous
?
-
-
-
?
4.2.1.84
methacrylonitrile + H2O
-
648439
Rhodococcus rhodochrous
methylacrylic acid amide
-
-
-
?
4.2.1.84
additional information
specificity overview
648439
Rhodococcus rhodochrous
?
-
-
-
-
4.2.1.84
additional information
specificity overview
648439
Rhodococcus rhodochrous J1
?
-
-
-
-
4.2.1.84
n-butyronitrile + H2O
-
648439
Rhodococcus rhodochrous
n-butyric acid amide
-
-
-
?
4.2.1.84
n-butyronitrile + H2O
-
648439
Rhodococcus rhodochrous J1
n-butyric acid amide
-
-
-
?
4.2.1.84
propionitrile + H2O
-
648439
Rhodococcus rhodochrous
propionic acid amide
-
-
-
?
4.2.1.84
propionitrile + H2O
-
648439
Rhodococcus rhodochrous J1
propionic acid amide
-
-
-
?
Subunits
EC Number
Subunits
Commentary
Organism
4.2.1.84
?
x * 26000, alpha, + x * 29000, beta, SDS-PAGE
Rhodococcus rhodochrous
Temperature Optimum [°C]
EC Number
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
4.2.1.84
35
-
-
Rhodococcus rhodochrous
Temperature Stability [°C]
EC Number
Temperature Stability Minimum [°C]
Temperature Stability Maximum [°C]
Commentary
Organism
4.2.1.84
50
-
stable up to, 30 min, pH 7.0, 44 mM n-butyric acid
Rhodococcus rhodochrous
pH Optimum
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
4.2.1.84
6.5
6.8
-
Rhodococcus rhodochrous
pH Range
EC Number
pH Minimum
pH Maximum
Commentary
Organism
4.2.1.84
5
8
about 50% of activity maximum at pH 5, about 60% of activity maximum at pH 8
Rhodococcus rhodochrous
pH Stability
EC Number
pH Stability
pH Stability Maximum
Commentary
Organism
4.2.1.84
6
9
20°C, 60 min, stable in this range, rapid loss of activity below pH 6.0 and above pH 9.0; 25°C, 10 min, stable in this range, rapid loss of activity below pH 6.0 and above pH 9.0
Rhodococcus rhodochrous
General Stability (protein specific)
EC Number
General Stability
Organism
4.2.1.84
organic acids stabilize, stable for more than 1 month in 0.1 M HEPES/KOH, pH 7.2, with 44 mM n-butyric acid, n-valeric acid, isovaleric acid, isobutyric acid or n-caproic acid
Rhodococcus rhodochrous
Inhibitors (protein specific)
EC Number
Inhibitors
Commentary
Organism
Structure
4.2.1.84
Ag+
0.01-1 mM AgNO3
Rhodococcus rhodochrous
4.2.1.84
diethyldithiocarbamate
0.1 mM
Rhodococcus rhodochrous
4.2.1.84
Hg2+
HgCl2, 0.01-1 mM
Rhodococcus rhodochrous
4.2.1.84
hydroxylamine
1 mM
Rhodococcus rhodochrous
4.2.1.84
KCN
0.01 mM
Rhodococcus rhodochrous
4.2.1.84
Na2S2O4
1 mM
Rhodococcus rhodochrous
4.2.1.84
phenylhydrazine
1 mM
Rhodococcus rhodochrous
4.2.1.84
Semicarbazide
1 mM, weak inhibition
Rhodococcus rhodochrous
KM Value [mM] (protein specific)
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
4.2.1.84
1.85
-
Isobutyronitrile
pH 7.0, 20ºC, H-NHase
Rhodococcus rhodochrous
4.2.1.84
1.89
-
acrylonitrile
pH 7.0, 20ºC, H-NHase
Rhodococcus rhodochrous
4.2.1.84
1.92
-
Propionitrile
pH 7.0, 20ºC, H-NHase
Rhodococcus rhodochrous
4.2.1.84
4.88
-
Crotononitrile
pH 7.0, 20ºC, H-NHase
Rhodococcus rhodochrous
4.2.1.84
6.76
-
Methacrylonitrile
pH 7.0, 20ºC
Rhodococcus rhodochrous
4.2.1.84
10
-
Cyanopyrazine
pH 7.0, 20ºC
Rhodococcus rhodochrous
4.2.1.84
12.2
-
Chloroacetonitrile
pH 7.0, 20ºC, H-NHase
Rhodococcus rhodochrous
4.2.1.84
18.5
-
4-cyanopyridine
pH 7.0, 20ºC, H-NHase
Rhodococcus rhodochrous
4.2.1.84
21.7
-
2-Cyanopyridine
pH 7.0, 20ºC, H-NHase
Rhodococcus rhodochrous
4.2.1.84
21.7
-
n-butyronitrile
pH 7.0, 20ºC, H-NHase
Rhodococcus rhodochrous
4.2.1.84
28.6
-
Benzonitrile
pH 7.0, 20ºC, H-NHase
Rhodococcus rhodochrous
4.2.1.84
200
-
3-Cyanopyridine
pH 7.0, 20ºC, H-NHase
Rhodococcus rhodochrous
Metals/Ions (protein specific)
EC Number
Metals/Ions
Commentary
Organism
Structure
4.2.1.84
Co2+
contains approximately 11-12 mol cobalt/mol enzyme
Rhodococcus rhodochrous
Molecular Weight [Da] (protein specific)
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
4.2.1.84
26000
-
x * 26000, alpha, + x * 29000, beta, SDS-PAGE
Rhodococcus rhodochrous
4.2.1.84
29000
-
x * 26000, alpha, + x * 29000, beta, SDS-PAGE
Rhodococcus rhodochrous
4.2.1.84
505000
530000
sedimentation equilibrium method, gel permeation HPLC
Rhodococcus rhodochrous
Purification (Commentary) (protein specific)
EC Number
Commentary
Organism
4.2.1.84
; ammonium sulfate fractionation, chromatography on DEAE-Sephacel, phenyl-Sepharose and Sephacryl S-300, 3.31fold purification
Rhodococcus rhodochrous
Specific Activity [micromol/min/mg] (protein specific)
EC Number
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
4.2.1.84
additional information
-
-
Rhodococcus rhodochrous
4.2.1.84
371
-
purified enzyme
Rhodococcus rhodochrous
Storage Stability (protein specific)
EC Number
Storage Stability
Organism
4.2.1.84
organic acids stabilize, stable for more than 1 month in 0.1 M HEPES/KOH, pH 7.2, with 44 mM n-butyric acid, n-valeric acid, isovaleric acid, isobutyric acid or n-caproic acid
Rhodococcus rhodochrous
Substrates and Products (Substrate) (protein specific)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
4.2.1.84
2-cyanopyridine + H2O
-
648439
Rhodococcus rhodochrous
pyridine-2-carbamide
-
-
-
?
4.2.1.84
3-cyanopyridine + H2O
-
648439
Rhodococcus rhodochrous
pyridine-3-carbamide
i.e. nicotinamide
648439
Rhodococcus rhodochrous
?
4.2.1.84
4-cyanopyridine + H2O
-
648439
Rhodococcus rhodochrous
pyridine-4-carbamide
-
-
-
?
4.2.1.84
acetonitrile + H2O
best substrate
648439
Rhodococcus rhodochrous
acetamide
-
-
-
?
4.2.1.84
acetonitrile + H2O
best substrate
648439
Rhodococcus rhodochrous J1
acetamide
-
-
-
?
4.2.1.84
acrylonitrile + H2O
-
648439
Rhodococcus rhodochrous
2-propenoic acid amide
i.e. acrylic acid amide
648439
Rhodococcus rhodochrous
?
4.2.1.84
benzonitrile + H2O
-
648439
Rhodococcus rhodochrous
benzoic acid amide
-
-
-
?
4.2.1.84
chloroacetonitrile + H2O
-
648439
Rhodococcus rhodochrous
chloroacetamide
-
-
-
?
4.2.1.84
chloroacetonitrile + H2O
-
648439
Rhodococcus rhodochrous J1
chloroacetamide
-
-
-
?
4.2.1.84
crotononitrile + H2O
19% of the activity with propionitrile
648439
Rhodococcus rhodochrous
(E)-2-butenoic acid amide
-
-
-
?
4.2.1.84
cyanopyrazine + H2O
-
648439
Rhodococcus rhodochrous
pyrazincarbamide
-
-
-
?
4.2.1.84
cyclopropylcyanide + H2O
-
648439
Rhodococcus rhodochrous
?
-
-
-
?
4.2.1.84
ethylene cyanhydrine + H2O
-
648439
Rhodococcus rhodochrous
?
-
-
-
?
4.2.1.84
methacrylonitrile + H2O
-
648439
Rhodococcus rhodochrous
methylacrylic acid amide
-
-
-
?
4.2.1.84
additional information
specificity overview
648439
Rhodococcus rhodochrous
?
-
-
-
-
4.2.1.84
additional information
specificity overview
648439
Rhodococcus rhodochrous J1
?
-
-
-
-
4.2.1.84
n-butyronitrile + H2O
-
648439
Rhodococcus rhodochrous
n-butyric acid amide
-
-
-
?
4.2.1.84
n-butyronitrile + H2O
-
648439
Rhodococcus rhodochrous J1
n-butyric acid amide
-
-
-
?
4.2.1.84
propionitrile + H2O
-
648439
Rhodococcus rhodochrous
propionic acid amide
-
-
-
?
4.2.1.84
propionitrile + H2O
-
648439
Rhodococcus rhodochrous J1
propionic acid amide
-
-
-
?
Subunits (protein specific)
EC Number
Subunits
Commentary
Organism
4.2.1.84
?
x * 26000, alpha, + x * 29000, beta, SDS-PAGE
Rhodococcus rhodochrous
Temperature Optimum [°C] (protein specific)
EC Number
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
4.2.1.84
35
-
-
Rhodococcus rhodochrous
Temperature Stability [°C] (protein specific)
EC Number
Temperature Stability Minimum [°C]
Temperature Stability Maximum [°C]
Commentary
Organism
4.2.1.84
50
-
stable up to, 30 min, pH 7.0, 44 mM n-butyric acid
Rhodococcus rhodochrous
pH Optimum (protein specific)
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
4.2.1.84
6.5
6.8
-
Rhodococcus rhodochrous
pH Range (protein specific)
EC Number
pH Minimum
pH Maximum
Commentary
Organism
4.2.1.84
5
8
about 50% of activity maximum at pH 5, about 60% of activity maximum at pH 8
Rhodococcus rhodochrous
pH Stability (protein specific)
EC Number
pH Stability
pH Stability Maximum
Commentary
Organism
4.2.1.84
6
9
20°C, 60 min, stable in this range, rapid loss of activity below pH 6.0 and above pH 9.0; 25°C, 10 min, stable in this range, rapid loss of activity below pH 6.0 and above pH 9.0
Rhodococcus rhodochrous