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Literature summary for 4.2.1.84 extracted from

  • Yamada, H.; Kobayashi, M.
    Nitrile hydratase and its application to industrial production of acrylamide (1996), Biosci. Biotechnol. Biochem., 60, 1391-1400.
    View publication on PubMed

Activating Compound

Activating Compound Comment Organism Structure
crotonamide induces high and low molecular mass forms of the enzyme Rhodococcus rhodochrous
cyclohexanecarboxamide induces the low molecular mass form of the enzyme Rhodococcus rhodochrous
light the enzyme activity is increased by exposure to near ultra-violet light Rhodococcus sp.
Urea induces the high molecular mass form of the enzyme Rhodococcus rhodochrous

Cloned(Commentary)

Cloned (Comment) Organism
-
Albifimbria verrucaria
expression in Escherichia coli Rhodococcus sp.
high molecular mass-NHase and low molecular mass-NHase genes cloned into Escherichia coli Rhodococcus rhodochrous

General Stability

General Stability Organism
unstable when diluted, but completely stabilized with low-molecular organic acids such as n-butyric acid, n-valeric acid, propionic acid and acetic acid Pseudomonas chlororaphis

Inhibitors

Inhibitors Comment Organism Structure
additional information high concentrations of acrylonitrile inhibit the enzyme activity Pseudomonas chlororaphis

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.001
-
phenylacetonitrile
-
Agrobacterium tumefaciens
0.0079
-
indole-3-acetonitrile
-
Agrobacterium tumefaciens
1.89
-
acrylonitrile
-
Rhodococcus rhodochrous

Metals/Ions

Metals/Ions Comment Organism Structure
Co2+
-
Rhodococcus sp.
Co2+ cannot replace ferrous or ferric ions Pseudomonas chlororaphis
Co2+ induces the enzyme activity Rhodococcus rhodochrous
Co2+ indispensable for high enzyme activity Rhodococcus rhodochrous
Fe2+
-
Rhodococcus rhodochrous
Fe2+ ferrous or ferric ions greatly increase the enzyme activity Pseudomonas chlororaphis
Fe2+ non-heme iron enzyme with a typical low-spin Fe(III)-active center Rhodococcus sp.
Fe2+ non-heme iron enzyme with a typical low-spin Fe(III)-active center Pseudomonas chlororaphis

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
27700
-
2 * 27700 Albifimbria verrucaria
130000
-
gel filtration, low molecular mass enzyme, L-NHase Rhodococcus rhodochrous
520000
-
high molecular mass enzyme, H-NHase Rhodococcus rhodochrous

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
indole-3-acetonitrile + H2O Agrobacterium tumefaciens
-
(indole-3-yl)acetamide
-
?
phenylacetonitrile + 2 H2O Agrobacterium tumefaciens
-
phenylacetic acid + NH3
-
?

Organism

Organism UniProt Comment Textmining
Agrobacterium tumefaciens
-
-
-
Albifimbria verrucaria
-
-
-
Arthrobacter sp.
-
-
-
Pseudomonas chlororaphis
-
-
-
Pseudomonas chlororaphis B23
-
-
-
Rhodococcus rhodochrous
-
-
-
Rhodococcus rhodochrous
-
J1
-
Rhodococcus rhodochrous J1
-
-
-
Rhodococcus rhodochrous J1
-
J1
-
Rhodococcus sp.
-
-
-

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
additional information
-
-
Rhodococcus rhodochrous
additional information
-
-
Rhodococcus sp.
additional information
-
141 U/mg of dry cells, Am 324 mutant strain Pseudomonas chlororaphis

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3-cyanopyridine + H2O
-
Rhodococcus sp. pyridine-3-carbamide
-
?
3-cyanopyridine + H2O
-
Rhodococcus rhodochrous pyridine-3-carbamide i.e. nicotinamide ?
acetonitrile + H2O
-
Rhodococcus rhodochrous acetamide
-
?
acetonitrile + H2O
-
Arthrobacter sp. acetamide
-
?
acetonitrile + H2O
-
Rhodococcus sp. acetamide
-
?
acetonitrile + H2O
-
Rhodococcus rhodochrous J1 acetamide
-
?
acrylonitrile + H2O
-
Rhodococcus rhodochrous 2-propenoic acid amide
-
?
acrylonitrile + H2O
-
Arthrobacter sp. 2-propenoic acid amide
-
?
acrylonitrile + H2O
-
Rhodococcus sp. 2-propenoic acid amide
-
?
acrylonitrile + H2O
-
Pseudomonas chlororaphis 2-propenoic acid amide
-
?
acrylonitrile + H2O
-
Pseudomonas chlororaphis B23 2-propenoic acid amide
-
?
acrylonitrile + H2O
-
Rhodococcus rhodochrous J1 2-propenoic acid amide
-
?
acrylonitrile + H2O
-
Rhodococcus rhodochrous acrylamide
-
?
cyanamide + H2O
-
Albifimbria verrucaria urea
-
?
hydroxyacetonitrile + H2O
-
Arthrobacter sp. hydroxyacetamide
-
?
indole-3-acetonitrile + H2O
-
Agrobacterium tumefaciens (indole-3-yl)acetamide
-
?
methacrylamide + H2O causes the greatest induction of activity Pseudomonas chlororaphis ?
-
?
methacrylamide + H2O causes the greatest induction of activity Pseudomonas chlororaphis B23 ?
-
?
additional information
-
Pseudomonas chlororaphis ?
-
?
additional information the enzyme acts on low-molecular aliphatic nitriles with 2-5 carbons but not on aliphatic nitriles with more than 6 carbons Arthrobacter sp. ?
-
?
additional information H-NHase acts preferentially on aliphatic nitriles, while L-NHase has a higher affinity for aromatic nitriles Rhodococcus rhodochrous ?
-
?
additional information
-
Pseudomonas chlororaphis B23 ?
-
?
additional information H-NHase acts preferentially on aliphatic nitriles, while L-NHase has a higher affinity for aromatic nitriles Rhodococcus rhodochrous J1 ?
-
?
n-butyronitrile + H2O
-
Rhodococcus rhodochrous n-butyric acid amide
-
?
n-butyronitrile + H2O
-
Arthrobacter sp. n-butyric acid amide
-
?
n-butyronitrile + H2O
-
Rhodococcus sp. n-butyric acid amide
-
?
n-butyronitrile + H2O
-
Pseudomonas chlororaphis n-butyric acid amide
-
?
n-butyronitrile + H2O
-
Pseudomonas chlororaphis B23 n-butyric acid amide
-
?
n-butyronitrile + H2O
-
Rhodococcus rhodochrous J1 n-butyric acid amide
-
?
phenylacetonitrile + 2 H2O
-
Agrobacterium tumefaciens phenylacetic acid + NH3
-
?
propionitrile + H2O
-
Rhodococcus rhodochrous propionic acid amide
-
?
propionitrile + H2O
-
Arthrobacter sp. propionic acid amide
-
?
propionitrile + H2O
-
Rhodococcus sp. propionic acid amide
-
?
propionitrile + H2O
-
Pseudomonas chlororaphis propionic acid amide
-
?
propionitrile + H2O
-
Pseudomonas chlororaphis B23 propionic acid amide
-
?
propionitrile + H2O
-
Rhodococcus rhodochrous J1 propionic acid amide
-
?

Subunits

Subunits Comment Organism
dimer
-
Rhodococcus sp.
dimer 2 * 27700 Albifimbria verrucaria

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
5
-
-
Rhodococcus rhodochrous

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
additional information
-
the enzyme is heat-stable Rhodococcus rhodochrous

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7
-
-
Rhodococcus rhodochrous

Cofactor

Cofactor Comment Organism Structure
pyrroloquinoline quinone
-
Pseudomonas chlororaphis