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Literature summary for 4.2.1.84 extracted from

  • Nagasawa, T.; Takeuchi, K.; Yamada, H.
    Characterization of a new cobalt-containing nitrile hydratase purified from urea-induced cells of Rhodococcus rhodochrous J1 (1991), Eur. J. Biochem., 196, 581-589.
    View publication on PubMed
Search for more literature for 4.2.1.84

General Stability

General Stability Organism
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

Inhibitors Comment Organism Structure
Ag+ 0.01-1 mM AgNO3 Rhodococcus rhodochrous
diethyldithiocarbamate 0.1 mM Rhodococcus rhodochrous
Hg2+ HgCl2, 0.01-1 mM Rhodococcus rhodochrous
hydroxylamine 1 mM Rhodococcus rhodochrous
KCN 0.01 mM Rhodococcus rhodochrous
Na2S2O4 1 mM Rhodococcus rhodochrous
phenylhydrazine 1 mM Rhodococcus rhodochrous
Semicarbazide 1 mM, weak inhibition Rhodococcus rhodochrous

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.85
-
 Isobutyronitrile pH 7.0, 20ºC, H-NHase Rhodococcus rhodochrous
1.89
-
 acrylonitrile pH 7.0, 20ºC, H-NHase Rhodococcus rhodochrous
1.92
-
 Propionitrile pH 7.0, 20ºC, H-NHase Rhodococcus rhodochrous
4.88
-
 Crotononitrile pH 7.0, 20ºC, H-NHase Rhodococcus rhodochrous
6.76
-
 Methacrylonitrile pH 7.0, 20ºC Rhodococcus rhodochrous
10
-
 Cyanopyrazine pH 7.0, 20ºC Rhodococcus rhodochrous
12.2
-
 Chloroacetonitrile pH 7.0, 20ºC, H-NHase Rhodococcus rhodochrous
18.5
-
 4-cyanopyridine pH 7.0, 20ºC, H-NHase Rhodococcus rhodochrous
21.7
-
 n-butyronitrile pH 7.0, 20ºC, H-NHase Rhodococcus rhodochrous
21.7
-
 2-Cyanopyridine pH 7.0, 20ºC, H-NHase Rhodococcus rhodochrous
28.6
-
 Benzonitrile pH 7.0, 20ºC, H-NHase Rhodococcus rhodochrous
200
-
 3-Cyanopyridine pH 7.0, 20ºC, H-NHase Rhodococcus rhodochrous

Metals/Ions

Metals/Ions Comment Organism Structure
Co2+ contains approximately 11-12 mol cobalt/mol enzyme Rhodococcus rhodochrous

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
26000
-
 x * 26000, alpha, + x * 29000, beta, SDS-PAGE Rhodococcus rhodochrous
29000
-
 x * 26000, alpha, + x * 29000, beta, SDS-PAGE Rhodococcus rhodochrous
505000 530000 sedimentation equilibrium method, gel permeation HPLC Rhodococcus rhodochrous

Organism

Organism UniProt Comment Textmining
Rhodococcus rhodochrous
-
 2 kinds of enzyme, a high molecular mass: H-NHase, and a low molecular mass enzyme: L-NHase
-
Rhodococcus rhodochrous J1
-
 2 kinds of enzyme, a high molecular mass: H-NHase, and a low molecular mass enzyme: L-NHase
-

Purification (Commentary)

Purification (Comment) Organism
-
 Rhodococcus rhodochrous
ammonium sulfate fractionation, chromatography on DEAE-Sephacel, phenyl-Sepharose and Sephacryl S-300, 3.31fold purification Rhodococcus rhodochrous

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
additional information
-
-
 Rhodococcus rhodochrous
371
-
 purified enzyme Rhodococcus rhodochrous

Storage Stability

Storage Stability Organism
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)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2-cyanopyridine + H2O
-
 Rhodococcus rhodochrous pyridine-2-carbamide
-
 ?
3-cyanopyridine + H2O
-
 Rhodococcus rhodochrous pyridine-3-carbamide i.e. nicotinamide ?
3-cyanopyridine + H2O
-
 Rhodococcus rhodochrous J1 pyridine-3-carbamide i.e. nicotinamide ?
4-cyanopyridine + H2O
-
 Rhodococcus rhodochrous isonicotinamide
-
 ?
acetonitrile + H2O best substrate Rhodococcus rhodochrous acetamide
-
 ?
acetonitrile + H2O best substrate Rhodococcus rhodochrous J1 acetamide
-
 ?
acrylonitrile + H2O
-
 Rhodococcus rhodochrous 2-propenoic acid amide i.e. acrylic acid amide ?
acrylonitrile + H2O
-
 Rhodococcus rhodochrous J1 2-propenoic acid amide i.e. acrylic acid amide ?
benzonitrile + H2O
-
 Rhodococcus rhodochrous benzoic acid amide
-
 ?
chloroacetonitrile + H2O
-
 Rhodococcus rhodochrous chloroacetamide
-
 ?
chloroacetonitrile + H2O
-
 Rhodococcus rhodochrous J1 chloroacetamide
-
 ?
crotononitrile + H2O 19% of the activity with propionitrile Rhodococcus rhodochrous (E)-2-butenoic acid amide
-
 ?
cyanopyrazine + H2O
-
 Rhodococcus rhodochrous pyrazincarbamide
-
 ?
cyclopropylcyanide + H2O
-
 Rhodococcus rhodochrous ?
-
 ?
ethylene cyanhydrine + H2O
-
 Rhodococcus rhodochrous ?
-
 ?
isobutyronitrile + H2O
-
 Rhodococcus rhodochrous isobutyramide
-
 ?
methacrylonitrile + H2O
-
 Rhodococcus rhodochrous methylacrylic acid amide
-
 ?
additional information specificity overview Rhodococcus rhodochrous ?
-
 ?
additional information specificity overview Rhodococcus rhodochrous J1 ?
-
 ?
n-butyronitrile + H2O
-
 Rhodococcus rhodochrous n-butyric acid amide
-
 ?
propionitrile + H2O
-
 Rhodococcus rhodochrous propionic acid amide
-
 ?

Subunits

Subunits Comment Organism
? x * 26000, alpha, + x * 29000, beta, SDS-PAGE Rhodococcus rhodochrous

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
35
-
-
 Rhodococcus rhodochrous

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
50
-
 stable up to, 30 min, pH 7.0, 44 mM n-butyric acid Rhodococcus rhodochrous

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
6.5 6.8
-
 Rhodococcus rhodochrous

pH Range

pH Minimum pH Maximum Comment Organism
5 8 about 50% of activity maximum at pH 5, about 60% of activity maximum at pH 8 Rhodococcus rhodochrous

pH Stability

pH Stability pH Stability Maximum Comment Organism
6 9 20°C, 60 min, stable in this range, rapid loss of activity below pH 6.0 and above pH 9.0 Rhodococcus rhodochrous
6 9 25°C, 10 min, stable in this range, rapid loss of activity below pH 6.0 and above pH 9.0 Rhodococcus rhodochrous