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Literature summary for 4.2.1.84 extracted from

  • Choi, Y.H.; Uhm, K.; Kim, H.
    Biochemical characterization of Rhodococcus erythropolis N'4 nitrile hydratase acting on 4-chloro-3-hydroxybutyronitrile (2008), J. Mol. Catal. B, 55, 157-163.
No PubMed abstract available
Search for more literature for 4.2.1.84

Application

Application Comment Organism
synthesis enzyme can be used in conjunction with a stereoselective amidase to synthesize ethyl (S)-4-chloro-3-hydroxybutyrate, an intermediate for a hypercholesterolemia drug, Atorvastatin Rhodococcus erythropolis

General Stability

General Stability Organism
the partially purified enzyme is stable in the presence of organic acids at higher temperatures Rhodococcus erythropolis

Inhibitors

Inhibitors Comment Organism Structure
1,10-phenanthroline 39% reduced activity at 1 mM Rhodococcus erythropolis
2-mercaptoethanol 9% reduced activity at 1 mM Rhodococcus erythropolis
ammonium persulfate 52% reduced activity at 1 mM Rhodococcus erythropolis
cobalt chloride 20% reduced activity at 1 mM Rhodococcus erythropolis
copper sulfate 95% reduced activity at 1 mM Rhodococcus erythropolis
dithiothreitol 18% reduced activity at 1 mM Rhodococcus erythropolis
EDTA 15% reduced activity at 1 mM Rhodococcus erythropolis
Ferrous sulfate 54% reduced activity at 1 mM Rhodococcus erythropolis
hydrogen peroxide 97% reduced activity at 1 mM Rhodococcus erythropolis
iodoacetamide 22% reduced activity at 1 mM Rhodococcus erythropolis
Mercury chloride 89% reduced activity at 1 mM Rhodococcus erythropolis
PMSF 83% reduced activity at 1 mM Rhodococcus erythropolis
Silver nitrate 82% reduced activity at 1 mM Rhodococcus erythropolis
zinc chloride 56% reduced activity at 1 mM Rhodococcus erythropolis

Metals/Ions

Metals/Ions Comment Organism Structure
Fe3+ NHase N'4 is a Fe-type enzyme with a Cys109-Ser110-Leu111-Cys112-Ser113-Cys114 sequence Rhodococcus erythropolis

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
22953
-
 2 * 27000, alpha-subunit, + 2 * 29000, beta-subunit, alpha2beta2 structure, SDS-PAGE, 2 * 22953, alpha-subunit, + 2 * 23486, beta-subunit, alpha2beta2 structure, sequence calculation Rhodococcus erythropolis
23486
-
 2 * 27000, alpha-subunit, + 2 * 29000, beta-subunit, alpha2beta2 structure, SDS-PAGE, 2 * 22953, alpha-subunit, + 2 * 23486, beta-subunit, alpha2beta2 structure, sequence calculation Rhodococcus erythropolis
27000
-
 2 * 27000, alpha-subunit, + 2 * 29000, beta-subunit, alpha2beta2 structure, SDS-PAGE, 2 * 22953, alpha-subunit, + 2 * 23486, beta-subunit, alpha2beta2 structure, sequence calculation Rhodococcus erythropolis
29000
-
 2 * 27000, alpha-subunit, + 2 * 29000, beta-subunit, alpha2beta2 structure, SDS-PAGE, 2 * 22953, alpha-subunit, + 2 * 23486, beta-subunit, alpha2beta2 structure, sequence calculation Rhodococcus erythropolis
92300
-
 gel filtration Rhodococcus erythropolis

Organism

Organism UniProt Comment Textmining
Rhodococcus erythropolis
-
-
-
Rhodococcus erythropolis N4
-
-
-

Purification (Commentary)

Purification (Comment) Organism
native enzyme 78fold by ammonium sulfate fractionation, anion exchange chromatography, and hydrophobic interaction chromatography Rhodococcus erythropolis

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
860
-
 partially purified enzyme Rhodococcus erythropolis

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
4-chloro-3-hydroxybutyronitrile + H2O the following reaction by an amidase leads to the correspondend carboxylic acid Rhodococcus erythropolis 4-chloro-3-hydroxybutyramide
-
 ?
acetonitrile + H2O very low activity Rhodococcus erythropolis acetamide
-
 ?
acetonitrile + H2O very low activity Rhodococcus erythropolis N4 acetamide
-
 ?
acrylonitrile + H2O
-
 Rhodococcus erythropolis 2-propenoic acid amide i.e. acrylamide ?
acrylonitrile + H2O
-
 Rhodococcus erythropolis N4 2-propenoic acid amide i.e. acrylamide ?
benzonitrile + H2O low activity Rhodococcus erythropolis benzoic acid amide
-
 ?
benzonitrile + H2O low activity Rhodococcus erythropolis N4 benzoic acid amide
-
 ?
butyronitrile + H2O best substrate Rhodococcus erythropolis butyric acid amide
-
 ?
isobutyronitrile + H2O
-
 Rhodococcus erythropolis isobutyric acid amide
-
 ?
isobutyronitrile + H2O
-
 Rhodococcus erythropolis N4 isobutyric acid amide
-
 ?
methacrylonitrile + H2O
-
 Rhodococcus erythropolis methacrylic acid amide
-
 ?
propionitrile + H2O
-
 Rhodococcus erythropolis propionic acid amide
-
 ?
propionitrile + H2O
-
 Rhodococcus erythropolis N4 propionic acid amide
-
 ?

Subunits

Subunits Comment Organism
tetramer 2 * 27000, alpha-subunit, + 2 * 29000, beta-subunit, alpha2beta2 structure, SDS-PAGE, 2 * 22953, alpha-subunit, + 2 * 23486, beta-subunit, alpha2beta2 structure, sequence calculation Rhodococcus erythropolis

Synonyms

Synonyms Comment Organism
NHase
-
 Rhodococcus erythropolis

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
25
-
-
 Rhodococcus erythropolis

Temperature Range [°C]

Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
15 35 15°C: about 60% of maximal activity, 35°C: about 55% of maximal activity, temperature profile Rhodococcus erythropolis

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
5 40 30 min, the purified enzyme is stable up to 25°C, inactivation at 40°C Rhodococcus erythropolis

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.5
-
-
 Rhodococcus erythropolis

pH Range

pH Minimum pH Maximum Comment Organism
7 8.5 pH 7.0: about 80% of maximal activity, pH 8.5: about 75% of maximal activity Rhodococcus erythropolis

pH Stability

pH Stability pH Stability Maximum Comment Organism
7 11 10°C, 30 min, purified enzyme, stable Rhodococcus erythropolis