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Literature summary 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

Application

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

Inhibitors

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

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
4.2.1.84 0.05
-
 Benzonitrile pH 7.0, 20ºC, L-NHase Rhodococcus rhodochrous
4.2.1.84 0.28
-
 4-cyanopyridine pH 7.0, 20ºC, L-NHase Rhodococcus rhodochrous
4.2.1.84 0.3
-
 3-Cyanopyridine pH 7.0, 20ºC, L-NHase Rhodococcus rhodochrous
4.2.1.84 0.44
-
 Methacrylonitrile pH 7.0, 20ºC, L-NHase Rhodococcus rhodochrous
4.2.1.84 0.56
-
 2-Cyanopyridine pH 7.0, 20ºC, L-NHase Rhodococcus rhodochrous
4.2.1.84 0.65
-
 n-butyronitrile pH 7.0, 20ºC, L-NHase Rhodococcus rhodochrous
4.2.1.84 0.71
-
 2-cyanopyrazine pH 7.0, 20ºC, L-NHase Rhodococcus rhodochrous
4.2.1.84 1.51
-
 Isobutyronitrile pH 7.0, 20ºC, L-NHase Rhodococcus rhodochrous
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 1.89
-
 Propionitrile pH 7.0, 20ºC, L-NHase Rhodococcus rhodochrous
4.2.1.84 1.92
-
 Propionitrile pH 7.0, 20ºC, H-NHase Rhodococcus rhodochrous
4.2.1.84 2.67
-
 acrylonitrile pH 7.0, 20ºC, L-NHase Rhodococcus rhodochrous
4.2.1.84 4.55
-
 Crotononitrile pH 7.0, 20ºC, L-NHase Rhodococcus rhodochrous
4.2.1.84 4.88
-
 Crotononitrile pH 7.0, 20ºC, H-NHase Rhodococcus rhodochrous
4.2.1.84 6.25
-
 Chloroacetonitrile pH 7.0, 20ºC, L-NHase Rhodococcus rhodochrous
4.2.1.84 6.76
-
 Methacrylonitrile pH 7.0, 20ºC, H-NHase Rhodococcus rhodochrous
4.2.1.84 10
-
 2-cyanopyrazine pH 7.0, 20ºC, H-NHase 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
-
 n-butyronitrile 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 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 Comment Organism Structure
4.2.1.84 Co2+ requirement, 1.7 mol cobalt/mol enzyme Rhodococcus rhodochrous

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
4.2.1.84 26000
-
 2 * 26000, alpha and 2 * 29000, beta, SDS-PAGE Rhodococcus rhodochrous
4.2.1.84 29000
-
 2 * 26000, alpha and 2 * 29000, beta, SDS-PAGE Rhodococcus rhodochrous
4.2.1.84 101000
-
 sedimentation equilibrium, low molecular mass enzyme, L-NHase Rhodococcus rhodochrous
4.2.1.84 130000
-
 gel filtration, low molecular mass enzyme, L-NHase Rhodococcus rhodochrous

Organism

EC Number Organism UniProt Comment 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 Purification (Comment) Organism
4.2.1.84 ammonium sulfate fractionation, chromatography on DEAE-Sephacel, phenyl-Sepharose CL-4B and gel filtration Rhodococcus rhodochrous

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
4.2.1.84 579
-
-
 Rhodococcus rhodochrous

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
4.2.1.84 2-cyanopyridine + H2O
-
 Rhodococcus rhodochrous pyridine-2-carbamide
-
 ?
4.2.1.84 3-cyanopyridine + H2O
-
 Rhodococcus rhodochrous pyridine-3-carbamide
-
 ?
4.2.1.84 4-cyanopyridine + H2O
-
 Rhodococcus rhodochrous isonicotinamide
-
 ?
4.2.1.84 acrylonitrile + H2O
-
 Rhodococcus rhodochrous 2-propenoic acid amide
-
 ?
4.2.1.84 acrylonitrile + H2O
-
 Rhodococcus rhodochrous J1 2-propenoic acid amide
-
 ?
4.2.1.84 benzonitrile + H2O
-
 Rhodococcus rhodochrous benzoic acid amide
-
 ?
4.2.1.84 chloroacetonitrile + H2O
-
 Rhodococcus rhodochrous chloroacetamide
-
 ?
4.2.1.84 chloroacetonitrile + H2O
-
 Rhodococcus rhodochrous J1 chloroacetamide
-
 ?
4.2.1.84 crotononitrile + H2O
-
 Rhodococcus rhodochrous (E)-2-butenoic acid amide
-
 ?
4.2.1.84 crotononitrile + H2O
-
 Rhodococcus rhodochrous J1 (E)-2-butenoic acid amide
-
 ?
4.2.1.84 cyanopyrazine + H2O
-
 Rhodococcus rhodochrous pyrazincarbamide
-
 ?
4.2.1.84 isobutyronitrile + H2O
-
 Rhodococcus rhodochrous isobutyramide
-
 ?
4.2.1.84 methacrylonitrile + H2O
-
 Rhodococcus rhodochrous methylacrylic acid amide
-
 ?
4.2.1.84 n-butyronitrile + H2O
-
 Rhodococcus rhodochrous n-butyric acid amide
-
 ?
4.2.1.84 n-butyronitrile + H2O
-
 Rhodococcus rhodochrous J1 n-butyric acid amide
-
 ?
4.2.1.84 propionitrile + H2O
-
 Rhodococcus rhodochrous propionic acid amide
-
 ?
4.2.1.84 propionitrile + H2O
-
 Rhodococcus rhodochrous J1 propionic acid amide
-
 ?

Subunits

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

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
4.2.1.84 40
-
 H-NHase Rhodococcus rhodochrous

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
4.2.1.84 50
-
 below, stable, H-NHase Rhodococcus rhodochrous

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
4.2.1.84 8.8
-
 H-NHase Rhodococcus rhodochrous

pH Range

EC Number pH Minimum pH Maximum Comment Organism
4.2.1.84 6 8.5 H-NHase Rhodococcus rhodochrous