Nitrile hydratase and its application to industrial production of acrylamide
Yamada, H.; Kobayashi, M.; Biosci. Biotechnol. Biochem. 60, 1391-1400 (1996) 
Data extracted from this reference:
Activating Compound
EC Number
Activating Compound
Commentary
Organism
Structure
4.2.1.84
crotonamide
induces high and low molecular mass forms of the enzyme
Rhodococcus rhodochrous
4.2.1.84
cyclohexanecarboxamide
induces the low molecular mass form of the enzyme
Rhodococcus rhodochrous
4.2.1.84
light
the enzyme activity is increased by exposure to near ultra-violet light
Rhodococcus sp.
4.2.1.84
Urea
induces the high molecular mass form of the enzyme
Rhodococcus rhodochrous
Cloned(Commentary)
EC Number
Cloned (Commentary)
Organism
4.2.1.84
-
Albifimbria verrucaria
4.2.1.84
expression in Escherichia coli
Rhodococcus sp.
4.2.1.84
high molecular mass-NHase and low molecular mass-NHase genes cloned into Escherichia coli
Rhodococcus rhodochrous
General Stability
EC Number
General Stability
Organism
4.2.1.84
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
EC Number
Inhibitors
Commentary
Organism
Structure
4.2.1.84
additional information
high concentrations of acrylonitrile inhibit the enzyme activity
Pseudomonas chlororaphis
KM Value [mM]
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
4.2.1.84
0.001
-
phenylacetonitrile
-
Agrobacterium tumefaciens
4.2.1.84
0.0079
-
indole-3-acetonitrile
-
Agrobacterium tumefaciens
4.2.1.84
1.89
-
acrylonitrile
-
Rhodococcus rhodochrous
Metals/Ions
EC Number
Metals/Ions
Commentary
Organism
Structure
4.2.1.84
Co2+
-
Rhodococcus sp.
4.2.1.84
Co2+
cannot replace ferrous or ferric ions
Pseudomonas chlororaphis
4.2.1.84
Co2+
indispensable for high enzyme activity; induces the enzyme activity
Rhodococcus rhodochrous
4.2.1.84
Fe2+
-
Rhodococcus rhodochrous
4.2.1.84
Fe2+
non-heme iron enzyme with a typical low-spin Fe(III)-active center
Rhodococcus sp.
4.2.1.84
Fe2+
ferrous or ferric ions greatly increase the enzyme activity; non-heme iron enzyme with a typical low-spin Fe(III)-active center
Pseudomonas chlororaphis
Molecular Weight [Da]
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
4.2.1.84
27700
-
2 * 27700
Albifimbria verrucaria
4.2.1.84
130000
-
gel filtration, low molecular mass enzyme, L-NHase
Rhodococcus rhodochrous
4.2.1.84
520000
-
high molecular mass enzyme, H-NHase
Rhodococcus rhodochrous
Natural Substrates/ Products (Substrates)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
ID
4.2.1.84
indole-3-acetonitrile + H2O
Agrobacterium tumefaciens
-
(indole-3-yl)acetamide
-
Agrobacterium tumefaciens
?
4.2.1.84
phenylacetonitrile + 2 H2O
Agrobacterium tumefaciens
-
phenylacetic acid + NH3
-
Agrobacterium tumefaciens
?
Organism
EC Number
Organism
UniProt
Commentary
Textmining
4.2.1.84
Agrobacterium tumefaciens
-
-
-
4.2.1.84
Albifimbria verrucaria
-
-
-
4.2.1.84
Arthrobacter sp.
-
-
-
4.2.1.84
Pseudomonas chlororaphis
-
-
-
4.2.1.84
Pseudomonas chlororaphis B23
-
-
-
4.2.1.84
Rhodococcus rhodochrous
-
; J1
-
4.2.1.84
Rhodococcus rhodochrous J1
-
; J1
-
4.2.1.84
Rhodococcus sp.
-
-
-
Specific Activity [micromol/min/mg]
EC Number
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
4.2.1.84
additional information
-
141 U/mg of dry cells, Am 324 mutant strain
Pseudomonas chlororaphis
4.2.1.84
additional information
-
-
Rhodococcus rhodochrous
4.2.1.84
additional information
-
-
Rhodococcus sp.
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
Substrate Product ID
4.2.1.84
3-cyanopyridine + H2O
-
648444
Rhodococcus sp.
pyridine-3-carbamide
-
648444
Rhodococcus sp.
-
4.2.1.84
3-cyanopyridine + H2O
-
648444
Rhodococcus rhodochrous
pyridine-3-carbamide
i.e. nicotinamide
648444
Rhodococcus rhodochrous
?
4.2.1.84
acetonitrile + H2O
-
648444
Rhodococcus rhodochrous
acetamide
-
-
-
?
4.2.1.84
acetonitrile + H2O
-
648444
Arthrobacter sp.
acetamide
-
-
-
-
4.2.1.84
acetonitrile + H2O
-
648444
Rhodococcus sp.
acetamide
-
-
-
?
4.2.1.84
acetonitrile + H2O
-
648444
Rhodococcus rhodochrous J1
acetamide
-
-
-
?
4.2.1.84
acrylonitrile + H2O
-
648444
Rhodococcus rhodochrous
2-propenoic acid amide
-
-
-
?
4.2.1.84
acrylonitrile + H2O
-
648444
Arthrobacter sp.
2-propenoic acid amide
-
-
-
-
4.2.1.84
acrylonitrile + H2O
-
648444
Rhodococcus sp.
2-propenoic acid amide
-
-
-
?
4.2.1.84
acrylonitrile + H2O
-
648444
Pseudomonas chlororaphis
2-propenoic acid amide
-
-
-
?
4.2.1.84
acrylonitrile + H2O
-
648444
Pseudomonas chlororaphis B23
2-propenoic acid amide
-
-
-
?
4.2.1.84
acrylonitrile + H2O
-
648444
Rhodococcus rhodochrous J1
2-propenoic acid amide
-
-
-
?
4.2.1.84
acrylonitrile + H2O
-
648444
Rhodococcus rhodochrous
acrylamide
-
-
-
-
4.2.1.84
cyanamide + H2O
-
648444
Albifimbria verrucaria
urea
-
648444
Albifimbria verrucaria
?
4.2.1.84
hydroxyacetonitrile + H2O
-
648444
Arthrobacter sp.
hydroxyacetamide
-
648444
Arthrobacter sp.
-
4.2.1.84
indole-3-acetonitrile + H2O
-
648444
Agrobacterium tumefaciens
(indole-3-yl)acetamide
-
648444
Agrobacterium tumefaciens
?
4.2.1.84
methacrylamide + H2O
causes the greatest induction of activity
648444
Pseudomonas chlororaphis
?
-
-
-
?
4.2.1.84
methacrylamide + H2O
causes the greatest induction of activity
648444
Pseudomonas chlororaphis B23
?
-
-
-
?
4.2.1.84
additional information
-
648444
Pseudomonas chlororaphis
?
-
-
-
-
4.2.1.84
additional information
the enzyme acts on low-molecular aliphatic nitriles with 2-5 carbons but not on aliphatic nitriles with more than 6 carbons
648444
Arthrobacter sp.
?
-
-
-
-
4.2.1.84
additional information
H-NHase acts preferentially on aliphatic nitriles, while L-NHase has a higher affinity for aromatic nitriles
648444
Rhodococcus rhodochrous
?
-
-
-
-
4.2.1.84
additional information
-
648444
Pseudomonas chlororaphis B23
?
-
-
-
-
4.2.1.84
additional information
H-NHase acts preferentially on aliphatic nitriles, while L-NHase has a higher affinity for aromatic nitriles
648444
Rhodococcus rhodochrous J1
?
-
-
-
-
4.2.1.84
n-butyronitrile + H2O
-
648444
Rhodococcus rhodochrous
n-butyric acid amide
-
-
-
-
4.2.1.84
n-butyronitrile + H2O
-
648444
Arthrobacter sp.
n-butyric acid amide
-
-
-
-
4.2.1.84
n-butyronitrile + H2O
-
648444
Rhodococcus sp.
n-butyric acid amide
-
-
-
-
4.2.1.84
n-butyronitrile + H2O
-
648444
Pseudomonas chlororaphis
n-butyric acid amide
-
-
-
-
4.2.1.84
n-butyronitrile + H2O
-
648444
Pseudomonas chlororaphis B23
n-butyric acid amide
-
-
-
-
4.2.1.84
n-butyronitrile + H2O
-
648444
Rhodococcus rhodochrous J1
n-butyric acid amide
-
-
-
-
4.2.1.84
phenylacetonitrile + 2 H2O
-
648444
Agrobacterium tumefaciens
phenylacetic acid + NH3
-
648444
Agrobacterium tumefaciens
?
4.2.1.84
propionitrile + H2O
-
648444
Rhodococcus rhodochrous
propionic acid amide
-
-
-
?
4.2.1.84
propionitrile + H2O
-
648444
Arthrobacter sp.
propionic acid amide
-
-
-
-
4.2.1.84
propionitrile + H2O
-
648444
Rhodococcus sp.
propionic acid amide
-
-
-
?
4.2.1.84
propionitrile + H2O
-
648444
Pseudomonas chlororaphis
propionic acid amide
-
-
-
?
4.2.1.84
propionitrile + H2O
-
648444
Pseudomonas chlororaphis B23
propionic acid amide
-
-
-
?
4.2.1.84
propionitrile + H2O
-
648444
Rhodococcus rhodochrous J1
propionic acid amide
-
-
-
?
Subunits
EC Number
Subunits
Commentary
Organism
4.2.1.84
dimer
-
Rhodococcus sp.
4.2.1.84
dimer
2 * 27700
Albifimbria verrucaria
Temperature Optimum [°C]
EC Number
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
4.2.1.84
5
-
-
Rhodococcus rhodochrous
Temperature Stability [°C]
EC Number
Temperature Stability Minimum [°C]
Temperature Stability Maximum [°C]
Commentary
Organism
4.2.1.84
additional information
-
the enzyme is heat-stable
Rhodococcus rhodochrous
pH Optimum
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
4.2.1.84
7
-
-
Rhodococcus rhodochrous
Cofactor
EC Number
Cofactor
Commentary
Organism
Structure
4.2.1.84
pyrroloquinoline quinone
-
Pseudomonas chlororaphis
Activating Compound (protein specific)
EC Number
Activating Compound
Commentary
Organism
Structure
4.2.1.84
crotonamide
induces high and low molecular mass forms of the enzyme
Rhodococcus rhodochrous
4.2.1.84
cyclohexanecarboxamide
induces the low molecular mass form of the enzyme
Rhodococcus rhodochrous
4.2.1.84
light
the enzyme activity is increased by exposure to near ultra-violet light
Rhodococcus sp.
4.2.1.84
Urea
induces the high molecular mass form of the enzyme
Rhodococcus rhodochrous
Cloned(Commentary) (protein specific)
EC Number
Commentary
Organism
4.2.1.84
-
Albifimbria verrucaria
4.2.1.84
expression in Escherichia coli
Rhodococcus sp.
4.2.1.84
high molecular mass-NHase and low molecular mass-NHase genes cloned into Escherichia coli
Rhodococcus rhodochrous
Cofactor (protein specific)
EC Number
Cofactor
Commentary
Organism
Structure
4.2.1.84
pyrroloquinoline quinone
-
Pseudomonas chlororaphis
General Stability (protein specific)
EC Number
General Stability
Organism
4.2.1.84
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 (protein specific)
EC Number
Inhibitors
Commentary
Organism
Structure
4.2.1.84
additional information
high concentrations of acrylonitrile inhibit the enzyme activity
Pseudomonas chlororaphis
KM Value [mM] (protein specific)
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
4.2.1.84
0.001
-
phenylacetonitrile
-
Agrobacterium tumefaciens
4.2.1.84
0.0079
-
indole-3-acetonitrile
-
Agrobacterium tumefaciens
4.2.1.84
1.89
-
acrylonitrile
-
Rhodococcus rhodochrous
Metals/Ions (protein specific)
EC Number
Metals/Ions
Commentary
Organism
Structure
4.2.1.84
Co2+
-
Rhodococcus sp.
4.2.1.84
Co2+
cannot replace ferrous or ferric ions
Pseudomonas chlororaphis
4.2.1.84
Co2+
indispensable for high enzyme activity; induces the enzyme activity
Rhodococcus rhodochrous
4.2.1.84
Fe2+
-
Rhodococcus rhodochrous
4.2.1.84
Fe2+
non-heme iron enzyme with a typical low-spin Fe(III)-active center
Rhodococcus sp.
4.2.1.84
Fe2+
ferrous or ferric ions greatly increase the enzyme activity; non-heme iron enzyme with a typical low-spin Fe(III)-active center
Pseudomonas chlororaphis
Molecular Weight [Da] (protein specific)
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
4.2.1.84
27700
-
2 * 27700
Albifimbria verrucaria
4.2.1.84
130000
-
gel filtration, low molecular mass enzyme, L-NHase
Rhodococcus rhodochrous
4.2.1.84
520000
-
high molecular mass enzyme, H-NHase
Rhodococcus rhodochrous
Natural Substrates/ Products (Substrates) (protein specific)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
ID
4.2.1.84
indole-3-acetonitrile + H2O
Agrobacterium tumefaciens
-
(indole-3-yl)acetamide
-
Agrobacterium tumefaciens
?
4.2.1.84
phenylacetonitrile + 2 H2O
Agrobacterium tumefaciens
-
phenylacetic acid + NH3
-
Agrobacterium tumefaciens
?
Specific Activity [micromol/min/mg] (protein specific)
EC Number
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
4.2.1.84
additional information
-
141 U/mg of dry cells, Am 324 mutant strain
Pseudomonas chlororaphis
4.2.1.84
additional information
-
-
Rhodococcus rhodochrous
4.2.1.84
additional information
-
-
Rhodococcus sp.
Substrates and Products (Substrate) (protein specific)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
ID
4.2.1.84
3-cyanopyridine + H2O
-
648444
Rhodococcus sp.
pyridine-3-carbamide
-
648444
Rhodococcus sp.
-
4.2.1.84
3-cyanopyridine + H2O
-
648444
Rhodococcus rhodochrous
pyridine-3-carbamide
i.e. nicotinamide
648444
Rhodococcus rhodochrous
?
4.2.1.84
acetonitrile + H2O
-
648444
Rhodococcus rhodochrous
acetamide
-
-
-
?
4.2.1.84
acetonitrile + H2O
-
648444
Arthrobacter sp.
acetamide
-
-
-
-
4.2.1.84
acetonitrile + H2O
-
648444
Rhodococcus sp.
acetamide
-
-
-
?
4.2.1.84
acetonitrile + H2O
-
648444
Rhodococcus rhodochrous J1
acetamide
-
-
-
?
4.2.1.84
acrylonitrile + H2O
-
648444
Rhodococcus rhodochrous
2-propenoic acid amide
-
-
-
?
4.2.1.84
acrylonitrile + H2O
-
648444
Arthrobacter sp.
2-propenoic acid amide
-
-
-
-
4.2.1.84
acrylonitrile + H2O
-
648444
Rhodococcus sp.
2-propenoic acid amide
-
-
-
?
4.2.1.84
acrylonitrile + H2O
-
648444
Pseudomonas chlororaphis
2-propenoic acid amide
-
-
-
?
4.2.1.84
acrylonitrile + H2O
-
648444
Pseudomonas chlororaphis B23
2-propenoic acid amide
-
-
-
?
4.2.1.84
acrylonitrile + H2O
-
648444
Rhodococcus rhodochrous J1
2-propenoic acid amide
-
-
-
?
4.2.1.84
acrylonitrile + H2O
-
648444
Rhodococcus rhodochrous
acrylamide
-
-
-
-
4.2.1.84
cyanamide + H2O
-
648444
Albifimbria verrucaria
urea
-
648444
Albifimbria verrucaria
?
4.2.1.84
hydroxyacetonitrile + H2O
-
648444
Arthrobacter sp.
hydroxyacetamide
-
648444
Arthrobacter sp.
-
4.2.1.84
indole-3-acetonitrile + H2O
-
648444
Agrobacterium tumefaciens
(indole-3-yl)acetamide
-
648444
Agrobacterium tumefaciens
?
4.2.1.84
methacrylamide + H2O
causes the greatest induction of activity
648444
Pseudomonas chlororaphis
?
-
-
-
?
4.2.1.84
methacrylamide + H2O
causes the greatest induction of activity
648444
Pseudomonas chlororaphis B23
?
-
-
-
?
4.2.1.84
additional information
-
648444
Pseudomonas chlororaphis
?
-
-
-
-
4.2.1.84
additional information
the enzyme acts on low-molecular aliphatic nitriles with 2-5 carbons but not on aliphatic nitriles with more than 6 carbons
648444
Arthrobacter sp.
?
-
-
-
-
4.2.1.84
additional information
H-NHase acts preferentially on aliphatic nitriles, while L-NHase has a higher affinity for aromatic nitriles
648444
Rhodococcus rhodochrous
?
-
-
-
-
4.2.1.84
additional information
-
648444
Pseudomonas chlororaphis B23
?
-
-
-
-
4.2.1.84
additional information
H-NHase acts preferentially on aliphatic nitriles, while L-NHase has a higher affinity for aromatic nitriles
648444
Rhodococcus rhodochrous J1
?
-
-
-
-
4.2.1.84
n-butyronitrile + H2O
-
648444
Rhodococcus rhodochrous
n-butyric acid amide
-
-
-
-
4.2.1.84
n-butyronitrile + H2O
-
648444
Arthrobacter sp.
n-butyric acid amide
-
-
-
-
4.2.1.84
n-butyronitrile + H2O
-
648444
Rhodococcus sp.
n-butyric acid amide
-
-
-
-
4.2.1.84
n-butyronitrile + H2O
-
648444
Pseudomonas chlororaphis
n-butyric acid amide
-
-
-
-
4.2.1.84
n-butyronitrile + H2O
-
648444
Pseudomonas chlororaphis B23
n-butyric acid amide
-
-
-
-
4.2.1.84
n-butyronitrile + H2O
-
648444
Rhodococcus rhodochrous J1
n-butyric acid amide
-
-
-
-
4.2.1.84
phenylacetonitrile + 2 H2O
-
648444
Agrobacterium tumefaciens
phenylacetic acid + NH3
-
648444
Agrobacterium tumefaciens
?
4.2.1.84
propionitrile + H2O
-
648444
Rhodococcus rhodochrous
propionic acid amide
-
-
-
?
4.2.1.84
propionitrile + H2O
-
648444
Arthrobacter sp.
propionic acid amide
-
-
-
-
4.2.1.84
propionitrile + H2O
-
648444
Rhodococcus sp.
propionic acid amide
-
-
-
?
4.2.1.84
propionitrile + H2O
-
648444
Pseudomonas chlororaphis
propionic acid amide
-
-
-
?
4.2.1.84
propionitrile + H2O
-
648444
Pseudomonas chlororaphis B23
propionic acid amide
-
-
-
?
4.2.1.84
propionitrile + H2O
-
648444
Rhodococcus rhodochrous J1
propionic acid amide
-
-
-
?
Subunits (protein specific)
EC Number
Subunits
Commentary
Organism
4.2.1.84
dimer
-
Rhodococcus sp.
4.2.1.84
dimer
2 * 27700
Albifimbria verrucaria
Temperature Optimum [°C] (protein specific)
EC Number
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
4.2.1.84
5
-
-
Rhodococcus rhodochrous
Temperature Stability [°C] (protein specific)
EC Number
Temperature Stability Minimum [°C]
Temperature Stability Maximum [°C]
Commentary
Organism
4.2.1.84
additional information
-
the enzyme is heat-stable
Rhodococcus rhodochrous
pH Optimum (protein specific)
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
4.2.1.84
7
-
-
Rhodococcus rhodochrous