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Literature summary for 4.2.1.84 extracted from

  • Wieser, M.; Takeuchi, K.; Wada, Y.; Yamada, H.; Nagasawa, T.
    Low-molecular-mass nitrile hydratase from Rhodococcus rhodochrous J1: purification, substrate specificity and comparison with the analogous high-molecular-mass enzyme (1998), FEMS Microbiol. Lett., 169, 17-22.
No PubMed abstract available
Search for more literature for 4.2.1.84

Application

Application Comment Organism
synthesis H-NHase is used in the industrial production of acrylamide and nicotinamide Rhodococcus rhodochrous

Inhibitors

Inhibitors Comment Organism Structure
Ag2SO4 total inhibition at 1 mM, L-NHase Rhodococcus rhodochrous
H2O2 1 mM, no effect on L-NHase activity Rhodococcus rhodochrous
Hg+ Hg2Cl2, 1 mM, total inhibition, L-NHase Rhodococcus rhodochrous

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.05
-
 Benzonitrile pH 7.0, 20ºC, L-NHase Rhodococcus rhodochrous
0.28
-
 4-cyanopyridine pH 7.0, 20ºC, L-NHase Rhodococcus rhodochrous
0.3
-
 3-Cyanopyridine pH 7.0, 20ºC, L-NHase Rhodococcus rhodochrous
0.44
-
 Methacrylonitrile pH 7.0, 20ºC, L-NHase Rhodococcus rhodochrous
0.56
-
 2-Cyanopyridine pH 7.0, 20ºC, L-NHase Rhodococcus rhodochrous
0.65
-
 n-butyronitrile pH 7.0, 20ºC, L-NHase Rhodococcus rhodochrous
0.71
-
 2-cyanopyrazine pH 7.0, 20ºC, L-NHase Rhodococcus rhodochrous
1.51
-
 Isobutyronitrile pH 7.0, 20ºC, L-NHase Rhodococcus rhodochrous
1.85
-
 Isobutyronitrile pH 7.0, 20ºC, H-NHase Rhodococcus rhodochrous
1.89
-
 acrylonitrile pH 7.0, 20ºC, H-NHase Rhodococcus rhodochrous
1.89
-
 Propionitrile pH 7.0, 20ºC, L-NHase Rhodococcus rhodochrous
1.92
-
 Propionitrile pH 7.0, 20ºC, H-NHase Rhodococcus rhodochrous
2.67
-
 acrylonitrile pH 7.0, 20ºC, L-NHase Rhodococcus rhodochrous
4.55
-
 Crotononitrile pH 7.0, 20ºC, L-NHase Rhodococcus rhodochrous
4.88
-
 Crotononitrile pH 7.0, 20ºC, H-NHase Rhodococcus rhodochrous
6.25
-
 Chloroacetonitrile pH 7.0, 20ºC, L-NHase Rhodococcus rhodochrous
6.76
-
 Methacrylonitrile pH 7.0, 20ºC, H-NHase Rhodococcus rhodochrous
10
-
 2-cyanopyrazine pH 7.0, 20ºC, H-NHase 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
-
 2-Cyanopyridine pH 7.0, 20ºC, H-NHase Rhodococcus rhodochrous
21.7
-
 n-butyronitrile 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+ requirement, 1.7 mol cobalt/mol enzyme Rhodococcus rhodochrous

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
26000
-
 2 * 26000, alpha and 2 * 29000, beta, SDS-PAGE Rhodococcus rhodochrous
29000
-
 2 * 26000, alpha and 2 * 29000, beta, SDS-PAGE Rhodococcus rhodochrous
101000
-
 sedimentation equilibrium, low molecular mass enzyme, L-NHase Rhodococcus rhodochrous
130000
-
 gel filtration, low molecular mass enzyme, L-NHase 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
ammonium sulfate fractionation, chromatography on DEAE-Sephacel, phenyl-Sepharose CL-4B and gel filtration Rhodococcus rhodochrous

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
579
-
-
 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
-
 ?
4-cyanopyridine + H2O
-
 Rhodococcus rhodochrous isonicotinamide
-
 ?
acrylonitrile + H2O
-
 Rhodococcus rhodochrous 2-propenoic acid amide
-
 ?
acrylonitrile + H2O
-
 Rhodococcus rhodochrous J1 2-propenoic acid amide
-
 ?
benzonitrile + H2O
-
 Rhodococcus rhodochrous benzoic acid amide
-
 ?
chloroacetonitrile + H2O
-
 Rhodococcus rhodochrous chloroacetamide
-
 ?
chloroacetonitrile + H2O
-
 Rhodococcus rhodochrous J1 chloroacetamide
-
 ?
crotononitrile + H2O
-
 Rhodococcus rhodochrous (E)-2-butenoic acid amide
-
 ?
crotononitrile + H2O
-
 Rhodococcus rhodochrous J1 (E)-2-butenoic acid amide
-
 ?
cyanopyrazine + H2O
-
 Rhodococcus rhodochrous pyrazincarbamide
-
 ?
isobutyronitrile + H2O
-
 Rhodococcus rhodochrous isobutyramide
-
 ?
methacrylonitrile + H2O
-
 Rhodococcus rhodochrous methylacrylic acid amide
-
 ?
n-butyronitrile + H2O
-
 Rhodococcus rhodochrous n-butyric acid amide
-
 ?
n-butyronitrile + H2O
-
 Rhodococcus rhodochrous J1 n-butyric acid amide
-
 ?
propionitrile + H2O
-
 Rhodococcus rhodochrous propionic acid amide
-
 ?
propionitrile + H2O
-
 Rhodococcus rhodochrous J1 propionic acid amide
-
 ?

Subunits

Subunits Comment Organism
tetramer 2 * 26000, alpha and 2 * 29000, beta, SDS-PAGE Rhodococcus rhodochrous

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
40
-
 H-NHase Rhodococcus rhodochrous

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
50
-
 below, stable, H-NHase Rhodococcus rhodochrous

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
8.8
-
 H-NHase Rhodococcus rhodochrous

pH Range

pH Minimum pH Maximum Comment Organism
6 8.5 H-NHase Rhodococcus rhodochrous