Information on EC 3.5.5.5 - Arylacetonitrilase

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The expected taxonomic range for this enzyme is: Bacteria, Eukaryota

EC NUMBER
COMMENTARY hide
3.5.5.5
-
RECOMMENDED NAME
GeneOntology No.
Arylacetonitrilase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
4-chlorophenylacetonitrile + 2 H2O = 4-chlorophenylacetate + NH3
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
hydrolysis
hydrolysis of nitriles
SYSTEMATIC NAME
IUBMB Comments
Arylacetonitrile aminohydrolase
Requires thiol compounds. Also hydrolyses other 4-substituted phenylacetonitriles, thien-2-ylacetonitrile, tolylacetonitriles, and, more slowly, benzyl cyanide.
CAS REGISTRY NUMBER
COMMENTARY hide
132053-06-6
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
-
UniProt
Manually annotated by BRENDA team
strain USDA 110
-
-
Manually annotated by BRENDA team
strain USDA 110
-
-
Manually annotated by BRENDA team
strain R1
-
-
Manually annotated by BRENDA team
strain R1
-
-
Manually annotated by BRENDA team
no activity in no activity in Pseudomonas syringae pv. tomato
strain DC3000
-
-
Manually annotated by BRENDA team
strain MTCC 5110
UniProt
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
evolution
metabolism
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
(2-bromophenyl)(hydroxy)acetonitrile + H2O
(2R)-(2-bromophenyl)(hydroxy)acetic acid + NH3
show the reaction diagram
-
74% activity compared to phenylacetonitrile
-
-
?
(2-fluorophenyl)(hydroxy)acetonitrile + H2O
(2R)-(2-fluorophenyl)(hydroxy)acetic acid + NH3
show the reaction diagram
-
439% activity compared to phenylacetonitrile
-
-
?
(3-bromophenyl)(hydroxy)acetonitrile + H2O
(2R)-(3-bromophenyl)(hydroxy)acetic acid + NH3
show the reaction diagram
-
706% activity compared to phenylacetonitrile
-
-
?
(3-chlorophenyl)(hydroxy)acetonitrile + H2O
(2R)-(3-chlorophenyl)(hydroxy)acetic acid + NH3
show the reaction diagram
-
810% activity compared to phenylacetonitrile
-
-
?
(3-fluorophenyl)(hydroxy)acetonitrile + H2O
(2R)-(3-fluorophenyl)(hydroxy)acetic acid + NH3
show the reaction diagram
-
454% activity compared to phenylacetonitrile
-
-
?
(4-bromophenyl)(hydroxy)acetonitrile + H2O
(2R)-(4-bromophenyl)(hydroxy)acetic acid + NH3
show the reaction diagram
-
680% activity compared to phenylacetonitrile
-
-
?
(4-chlorophenyl)(hydroxy)acetonitrile + H2O
(2R)-(4-chlorophenyl)(hydroxy)acetic acid + NH3
show the reaction diagram
-
1083% activity compared to phenylacetonitrile
-
-
?
(4-fluorophenyl)(hydroxy)acetonitrile + H2O
(2R)-(4-fluorophenyl)(hydroxy)acetic acid + NH3
show the reaction diagram
-
891% activity compared to phenylacetonitrile
-
-
?
(R,S)-2-phenylpropionitrile + H2O
(S)-2-phenylpropionic acid + NH3
show the reaction diagram
(R,S)-2-phenylpropionitrile + H2O
2-phenylpropionic acid + 2-phenylpropionamide + NH3
show the reaction diagram
(R,S)-2-phenylpropionitrile + H2O
2-phenylpropionic acid + NH3
show the reaction diagram
(R,S)-mandelonitrile + 2 H2O
(R)-(-)-mandelic acid + NH3
show the reaction diagram
-
-
-
-
?
(R,S)-mandelonitrile + 2 H2O
(R)-mandelic acid + NH3
show the reaction diagram
(R,S)-mandelonitrile + H2O
(R)-(-)mandelic acid + NH3
show the reaction diagram
(R,S)-mandelonitrile + H2O
(R)-mandelic acid + NH3
show the reaction diagram
(R,S)-mandelonitrile + H2O
mandelic acid + NH3
show the reaction diagram
(S)-mandelonitrile + 2 H2O
(S)-mandelic acid + NH3
show the reaction diagram
(S)-mandelonitrile + H2O
(S)-mandelic acid + NH3
show the reaction diagram
1,2-phenylenediacetonitrile + H2O
1,2-phenylenediacetic acid + NH3
show the reaction diagram
-
1.87% activity compared to phenylacetonitrile
-
-
?
1,4-dicyanobutane + H2O
adipic acid + NH3
show the reaction diagram
-
47% activity compared to phenylacetonitrile
-
-
?
1-cyclohexenylacetonitrile + H2O
1-cyclohexenylacetate + NH3
show the reaction diagram
-
at 3.3% the rate of 4-chlorobenzylcyanide hydrolysis
-
-
-
2 (R,S)-mandelonitrile + 2 H2O
(R)-mandelic acid + (S)-mandeloamide + NH3
show the reaction diagram
2,4-dichlorobenzylcyanide + H2O
2,4-dichlorophenylacetic acid + NH3
show the reaction diagram
-
at 6.5% the rate of 4-chlorobenzylcyanide hydrolysis
-
-
-
2-acetyloxy-2-methylphenylacetonitrile + 2 H2O
2-acetyloxy-2-methylphenylacetic acid + NH3
show the reaction diagram
2-acetyloxy-2-methylphenylacetonitrile + H2O
2-acetyloxy-2-methylphenylacetic acid + NH3
show the reaction diagram
-
-
-
-
?
2-chlorobenzylcyanide + H2O
2-chlorophenylacetic acid + NH3
show the reaction diagram
-
at 6% the rate of 4-chlorobenzylcyanide hydrolysis
-
-
-
2-chloromandelonitrile + H2O
(R)-2-chloromandelic acid + H2O
show the reaction diagram
-
33.75% activity compared to phenylacetonitrile
enantioselectivity ee 98.2%
-
?
2-chloromandelonitrile + H2O
?
show the reaction diagram
-
-
-
-
?
2-chlorophenyl acetonitrile + H2O
2-chlorobenzoic acid + NH3
show the reaction diagram
-
76.1% of the activity with mandelonitrile
-
-
ir
2-cyanopyridine + H2O
?
show the reaction diagram
-
101% activity compared to phenylacetonitrile
-
-
?
2-hydroxy-2-phenylacetonitrile + 2 H2O
2-hydroxy-2-phenylacetate + NH3
show the reaction diagram
2-hydroxy-2-phenylpropionitrile + H2O
2-hydroxy-2-phenylpropionic acid + 2-hydroxy-2-phenylpropionamide + NH3
show the reaction diagram
-
i.e. acetophenone cyanohydrin
product ratio of about 3.4:1 for 2-hydroxy-2-phenylpropionic acid and 2-hydroxy-2-phenylpropionamide
-
?
2-hydroxy-3-butenenitrile + H2O
2-hydroxy-3-butenoic acid + NH3
show the reaction diagram
-
-
-
-
?
2-methoxybenzylcyanide + H2O
2-methoxyphenylacetic acid + NH3
show the reaction diagram
-
at 2% the rate of 4-chlorobenzylcyanide hydrolysis
-
-
-
2-methoxyphenyl acetonitrile + H2O
2-methoxybenzoic acid + NH3
show the reaction diagram
-
44.5% of the activity with mandelonitrile
-
-
ir
2-methoxyphenylacetonitrile + 2 H2O
2-methoxyphenylacetic acid + NH3
show the reaction diagram
2-methyl-2-phenylpropionitrile + H2O
2-methyl-2-phenylpropionic acid + NH3
show the reaction diagram
-
-
-
-
?
2-methylglutaronitrile + H2O
2-methylglutaric acid + NH3
show the reaction diagram
-
0.12% activity compared to phenylacetonitrile
-
-
?
2-methylglutaronitrile + H2O
? + NH3
show the reaction diagram
-
63% activity compared to phenylacetonitrile
-
-
?
2-phenylbutyronitrile + 2 H2O
2-phenylbutyric acid + NH3
show the reaction diagram
2-phenylbutyronitrile + H2O
2-phenylbutyric acid + NH3
show the reaction diagram
-
0.2% activity compared to phenylacetonitrile
-
-
?
2-phenylglycinonitrile + H2O
2-phenylalanine + NH3
show the reaction diagram
-
80% of the activity with mandelonitrile. 35% enantiomeric excess for S-configuration in acid formation, 10.26% of product is amide with 92% enantiomeric excess for S-configuration
-
-
?
2-phenylglycinonitrile + H2O
2-phenylglycine + NH3
show the reaction diagram
-
3% activity compared to phenylacetonitrile
-
-
?
2-phenylpropionitrile + 2 H2O
2-phenylpropionic acid + NH3
show the reaction diagram
2-phenylpropionitrile + H2O
2-phenylpropionate + NH3
show the reaction diagram
-
low activity, 2% activity compared to phenylacetonitrile
-
?
2-phenylpropionitrile + H2O
2-phenylpropionic acid + NH3
show the reaction diagram
2-phenylvaleronitrile + 2 H2O
2-phenylvaleric acid + NH3
show the reaction diagram
-
-
-
-
?
2-phenylvaleronitrile + H2O
2-phenylvaleric acid + NH3
show the reaction diagram
-
700% of the activity with mandelonitrile
-
-
?
2-thiophene acetonitrile + H2O
thiophene-2-carboxylic acid + NH3
show the reaction diagram
-
137% of the activity with mandelonitrile
-
-
ir
2-thiopheneacetonitrile + H2O
2-thiopheneacetic acid + NH3
show the reaction diagram
2-trimethylsilyloxy-2-phenylacetonitrile + H2O
2-trimethylsilyloxy-2-phenylacetic acid + NH3
show the reaction diagram
-
4% activity compared to phenylacetonitrile
-
-
?
3,4-(methylenedioxy)phenylacetonitrile + H2O
3,4-(methylendioxy)phenylacetic acid + NH3
show the reaction diagram
-
at 65% the rate of 4-chlorobenzylcyanide hydrolysis
-
-
-
3,4-dichlorobenzylcyanide + H2O
3,4-dichlorophenylacetic acid + NH3
show the reaction diagram
-
at 14% the rate of 4-chlorobenzylcyanide hydrolysis
-
-
-
3,4-dimethoxyphenylacetonitrile + H2O
?
show the reaction diagram
-
-
-
-
?
3-aminopropionitrile + H2O
3-aminopropanoic acid + NH3
show the reaction diagram
-
7% activity compared to phenylacetonitrile
-
-
?
3-chlorobenzylcyanide + H2O
3-chlorophenylacetic acid + NH3
show the reaction diagram
-
at 43% the rate of 4-chlorobenzylcyanide hydrolysis
-
-
-
3-cyanopyridine + H2O
?
show the reaction diagram
-
43.2% activity compared to phenylacetonitrile
-
-
?
3-cyanopyridine + H2O
nicotinic acid + NH3
show the reaction diagram
3-hydroxyglutaronitrile + H2O
3-hydroxyglutaric acid + NH3
show the reaction diagram
-
0.12% activity compared to phenylacetonitrile
-
-
?
3-hydroxyphenylpropionitrile + H2O
3-hydroxyphenylpropionic acid + NH3
show the reaction diagram
-
0.12% activity compared to phenylacetonitrile
-
-
?
3-hydroxypropionitrile + H2O
3-hydroxypropanoate + NH3
show the reaction diagram
3-hydroxypropionitrile + H2O
3-hydroxypropionic acid + NH3
show the reaction diagram
-
0.07% activity compared to phenylacetonitrile
-
-
?
3-indoleacetonitrile + H2O
3-indoleacetic acid + NH3
show the reaction diagram
3-indolylacetonitrile + H2O
3-indolylacetic acid + NH3
show the reaction diagram
-
2% activity compared to phenylacetonitrile
-
-
?
3-methoxyphenylacetonitrile + 2 H2O
3-methoxyphenylacetic acid + NH3
show the reaction diagram
-
-
-
-
?
3-phenylpropionitrile + H2O
3-phenylpropionic acid + NH3
show the reaction diagram
3-pyridineacetonitrile + H2O
?
show the reaction diagram
3-thiopheneacetonitrile + H2O
3-thiopheneacetic acid + NH3
show the reaction diagram
4-aminobenzylcyanide + H2O
4-aminophenylacetic acid + NH3
show the reaction diagram
4-aminophenyl acetonitrile + H2O
4-aminobenzoic acid + NH3
show the reaction diagram
-
502.3% of the activity with mandelonitrile
-
-
ir
4-bromobenzylcyanide + H2O
?
show the reaction diagram
4-chlorobenzylcyanide + H2O
4-chlorophenylacetic acid + NH3
show the reaction diagram
4-chlorobutyronitrile + H2O
4-chlorobutyrate + NH3
show the reaction diagram
4-chlorobutyronitrile + H2O
4-chlorobutyric acid + NH3
show the reaction diagram
4-chlorophenyl acetonitrile + H2O
4-chlorobenzoic acid + NH3
show the reaction diagram
-
360.3% of the activity with mandelonitrile
-
-
ir
4-cyanopyridine + H2O
4-pyridinecarboxylic acid + NH3
show the reaction diagram
-
512% of the rate with benzonitrile, cell extract
-
-
?
4-cyanopyridine + H2O
?
show the reaction diagram
-
142% activity compared to phenylacetonitrile
-
-
?
4-fluorobenzylcyanide + H2O
4-fluorophenylacetic acid + NH3
show the reaction diagram
4-hydroxy benzonitrile + H2O
4-hydroxybenzoate + NH3
show the reaction diagram
-
23.1% of the activity with mandelonitrile
-
-
ir
4-hydroxybenzylcyanide + H2O
4-hydroxyphenylacetic acid + NH3
show the reaction diagram
-
at 29% the rate of 4-chlorobenzylcyanide hydrolysis
-
-
-
4-hydroxycinnamonitrile + H2O
4-hydroxycinnamic acid + NH3
show the reaction diagram
-
431% activity compared to phenylacetonitrile
-
-
?
4-hydroxyphenyl acetonitrile + H2O
4-hydroxybenzoic acid + NH3
show the reaction diagram
-
435% of the activity with mandelonitrile
-
-
ir
4-hydroxyphenylacetonitrile + H2O
4-hydroxyphenylacetate + NH3
show the reaction diagram
-
best substrate
-
-
?
4-methoxybenzylcyanide + H2O
4-methoxyphenylacetic acid + NH3
show the reaction diagram
-
at 62% the rate of 4-chlorobenzylcyanide hydrolysis
-
-
-
4-methoxyphenylacetonitrile + 2 H2O
4-methoxyphenylacetic acid + NH3
show the reaction diagram
-
-
-
-
?
4-methoxyphenylacetonitrile + H2O
4-methoxyphenylacetate + NH3
show the reaction diagram
-
81% of the activity with 4-hydroxyphenylacetonitrile
-
-
?
4-nitrobenzylcyanide + H2O
4-nitrophenylacetate + NH3
show the reaction diagram
4-phenylbutyronitrile + H2O
4-phenylbutyrate + NH3
show the reaction diagram
-
86% activity compared to phenylacetonitrile
-
-
?
acrylonitrile + H2O
acrylic acid + NH3
show the reaction diagram
acrylonitrile + H2O
propenoic acid + NH3
show the reaction diagram
adiponitrile + H2O
adipic acid + NH3
show the reaction diagram
-
17.2% of the activity with mandelonitrile
-
-
ir
allyl cyanide + NH3
3-butenoic acid + NH3
show the reaction diagram
-
11% activity compared to phenylacetonitrile
-
-
?
alpha-methylphenylacetonitrile + H2O
alpha-methylphenylacetic acid + NH3
show the reaction diagram
-
0.14% activity compared to phenylacetonitrile
-
-
?
Arylacetonitrile + H2O
?
show the reaction diagram
-
pathway in nitrile catabolism, inducible enzyme
-
-
-
benzonitrile + H2O
benzoic acid + NH3
show the reaction diagram
benzonitrile + H2O
crotonic acid + NH3
show the reaction diagram
-
1% activity compared to phenylacetonitrile
-
-
?
benzylcyanide + H2O
phenylacetic acid + NH3
show the reaction diagram
cinnamonitrile + H2O
cinnamic acid + NH3
show the reaction diagram
-
0.07% activity compared to phenylacetonitrile
-
-
?
crotononitrile + H2O
crotonic acid + NH3
show the reaction diagram
-
4% activity compared to phenylacetonitrile
-
-
?
cyclohexane carbonitrile + H2O
cyclohexlycarboxylic acid + NH3
show the reaction diagram
-
12.5% of the activity with mandelonitrile
-
-
ir
diphenylacetonitrile + H2O
diphenylacetic acid + NH3
show the reaction diagram
dodecanenitrile + H2O
dodecanoic acid + NH3
show the reaction diagram
-
0.04% activity compared to phenylacetonitrile
-
-
?
fumaronitrile + H2O
fumaric acid + NH3
show the reaction diagram
glycolonitrile + H2O
glycolic acid + NH3
show the reaction diagram
-
0.08% activity compared to phenylacetonitrile
-
-
?
heptanenitrile + H2O
heptanoic acid + NH3
show the reaction diagram
-
107% activity compared to phenylacetonitrile
-
-
?
hex-5-enenitrile + H2O
hex-5-enoic acid + NH3
show the reaction diagram
-
7.9% of the activity with mandelonitrile
-
-
ir
hexanenitrile + H2O
hexanoic acid + NH3
show the reaction diagram
-
60% activity compared to phenylacetonitrile
-
-
?
hydroxy(2-methoxyphenyl)acetonitrile + H2O
(2R)-hydroxy(2-methoxyphenyl)acetic acid + NH3
show the reaction diagram
-
178% activity compared to phenylacetonitrile
-
-
?
hydroxy(2-methylphenyl)acetonitrile + H2O
(2R)-hydroxy(2-methylphenyl)acetic acid + NH3
show the reaction diagram
-
194% activity compared to phenylacetonitrile
-
-
?
hydroxy(phenyl)acetonitrile + H2O
(2R)-hydroxy(phenyl)acetic acid + NH3
show the reaction diagram
-
404% activity compared to phenylacetonitrile
-
-
?
iminodiacetonitrile + H2O
iminodicaetic acid + NH3
show the reaction diagram
-
0.05% activity compared to phenylacetonitrile
-
-
?
indole 3-acetonitrile + H2O
(indol-3-yl)acetic acid + NH3
show the reaction diagram
-
319.9% of the activity with mandelonitrile
-
-
ir
indole-3-acetonitrile + H2O
indole-3-acetic acid + NH3
show the reaction diagram
isovaleronitrile + H2O
isovaleric acid + NH3
show the reaction diagram
-
11.4% of the activity with mandelonitrile
-
-
ir
m-tolylacetonitrile + H2O
m-tolylacetic acid + NH3
show the reaction diagram
m-xylenedicyanide + H2O
?
show the reaction diagram
-
at 2.7% the rate of 4-chlorobenzylcyanide hydrolysis
-
-
-
malononitrile + H2O
malonic acid + NH3
show the reaction diagram
mandelonitrile + 2 H2O
(R)-mandelic acid + NH3
show the reaction diagram
mandelonitrile + 2 H2O
2-hydroxyphenylacetic acid + NH3
show the reaction diagram
mandelonitrile + 2 H2O
mandelate + NH3
show the reaction diagram
mandelonitrile + 2 H2O
mandelic acid + NH3
show the reaction diagram
mandelonitrile + H2O
mandelic acid + NH3
show the reaction diagram
methylthioacetonitrile + H2O
methylthioacetic acid + NH3
show the reaction diagram
-
72% activity compared to phenylacetonitrile
-
-
?
n-butyronitrile + H2O
n-butyric acid + NH3
show the reaction diagram
o-tolylacetonitrile + H2O
o-tolylacetic acid + NH3
show the reaction diagram
octanedinitrile + H2O
? + NH3
show the reaction diagram
-
0.14% activity compared to phenylacetonitrile
-
-
?
p-tolylacetonitrile + H2O
p-tolylacetic acid + NH3
show the reaction diagram
-
at 72% the rate of 4-chlorobenzylcyanide hydrolysis
-
-
phenoxy acetonitrile + H2O
phenoxyacetate + NH3
show the reaction diagram
-
232.6% of the activity with mandelonitrile
-
-
ir
phenyl acetonitrile + H2O
benzoic acid + NH3
show the reaction diagram
-
342.3% of the activity with mandelonitrile
-
-
ir
phenyl glycinenitrile + H2O
phenylalanine + NH3
show the reaction diagram
-
192.5% of the activity with mandelonitrile
-
-
ir
phenyl thioacetonitrile + H2O
thiophenylacetate + NH3
show the reaction diagram
-
187.7% of the activity with mandelonitrile
-
-
ir
phenylacetonitrile + 2 H2O
phenylacetate + NH3
show the reaction diagram
phenylacetonitrile + 2 H2O
phenylacetic acid + NH3
show the reaction diagram
phenylacetonitrile + H2O
?
show the reaction diagram
-
-
-
-
?
phenylacetonitrile + H2O
phenylacetate + NH3
show the reaction diagram
phenylacetonitrile + H2O
phenylacetic acid + NH3
show the reaction diagram
phenylpropionitrile + 2 H2O
phenylpropionate + NH3
show the reaction diagram
phenylpropionitrile + H2O
phenylpropionic acid + NH3
show the reaction diagram
rac 2-(methoxy)-mandelonitrile + H2O
(R)-2-(methoxy)-mandelic acid + (S)-2-(methoxy)-mandelic acid + NH3
show the reaction diagram
-
-
10% activity compared to phenylacetonitrile, 92% of the product is the (R)-enantiomer when the overall conversion rate is 50%, at 85% overall conversion, the level of formed (R)-enantiomer is 27%
?
succinonitrile + H2O
succinic acid + NH3
show the reaction diagram
-
0.12% activity compared to phenylacetonitrile
-
-
?
valeronitrile + H2O
valeric acid + NH3
show the reaction diagram
additional information
?
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
2-acetyloxy-2-methylphenylacetonitrile + 2 H2O
2-acetyloxy-2-methylphenylacetic acid + NH3
show the reaction diagram
2-chloromandelonitrile + H2O
?
show the reaction diagram
-
-
-
-
?
3,4-dimethoxyphenylacetonitrile + H2O
?
show the reaction diagram
-
-
-
-
?
Arylacetonitrile + H2O
?
show the reaction diagram
-
pathway in nitrile catabolism, inducible enzyme
-
-
-
mandelonitrile + 2 H2O
(R)-mandelic acid + NH3
show the reaction diagram
mandelonitrile + 2 H2O
mandelate + NH3
show the reaction diagram
mandelonitrile + 2 H2O
mandelic acid + NH3
show the reaction diagram
-
-
-
-
?
phenylacetonitrile + 2 H2O
phenylacetate + NH3
show the reaction diagram
phenylacetonitrile + H2O
?
show the reaction diagram
-
-
-
-
?
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Mg2+
-
stimulates hydrolytic activity to 122%
additional information
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2,2'-dipyridyl
-
weak
3-Methyl-2-benzothiazolinone hydrazone
-
weak
8-hydroxyquinoline
-
weak
Ag2+
-
strong
ascorbic acid
-
-
Ba2+
-
8% inhibition at 5 mM
Ca2+
about 50% inhibition at 1 mM
Cs+
about 50% inhibition at 1 mM
D-cycloserine
-
weak
diethyldithiocarbamate
-
-
Disodium 4,5-dihydroxy-m-benzene-disulfonate
-
weak
dithiothreitol
about 50% inhibition at 1 mM
DTNB
-
strong
hydroxylamine
-
weak
iodoacetate
-
weak
L-cysteine
-
-
Li+
-
11% inhibition at 5 mM
Mg2+
91% inhibition at 1 mM
N-ethylmaleimide
-
-
p-chloromercuribenzoate
-
strong
phenylhydrazine
-
-
Semicarbazide
-
weak
additional information
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2-mercaptoethanol
dithiothreitol
-
activation
phenylacetonitrile
-
absolutely required as growth nitrogen source substrate for induction of the enzyme, cannot be substituted by ammonia or epsilon-caprolactam
Sodium hydrosulfide
-
activation
additional information
-
expression is induced by growing the strain in the presence of different nitriles and /or complex or inorganic nitrogen sources. The highest nitrile hydrolysing activity is observed with cells grown with 2-cyanopyridine and NaNO3
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.015
2-Chlorobenzylcyanide
-
-
0.38
2-chloromandelonitrile
-
at pH 7.0 and 30°C
3.4 - 11.4
2-hydroxy-2-phenylacetonitrile
0.091
2-Thiopheneacetonitrile
-
-
0.065
3-Chlorobenzylcyanide
-
-
0.009
3-Indoleacetonitrile
-
-
0.016
3-Thiopheneacetonitrile
-
-
0.023
3-tolylacetonitrile
-
-
0.022
4-Aminobenzylcyanide
-
-
0.333
4-Chlorobenzylcyanide
-
-
0.13
4-Fluorobenzylcyanide
-
-
0.105
4-Nitrobenzylcyanide
-
-
0.123
4-tolylacetonitrile
-
-
0.66
acrylonitrile
-
-
0.01
benzylcyanide
-
-
5.31
indole 3-acetonitrile
-
35°C, pH 7.5
4.74 - 13.4
mandelonitrile
3.61
phenyl acetonitrile
-
35°C, pH 7.5
0.065 - 4.36
phenylacetonitrile
0.8 - 1.3
phenylpropionitrile
0.087
rac 2-(methoxy)-mandelonitrile
-
pH 7.4, 30°C
additional information
additional information
-
kinetic and chiral analysis, overview
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
13.4
indole 3-acetonitrile
Pseudomonas putida
-
35°C, pH 7.5
11.8 - 26
mandelonitrile
1.1 - 18.85
phenyl acetonitrile
1.85
phenylacetonitrile
Bradyrhizobium japonicum
-
-
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.44
pH 8.0, in supernatant of the recombinant Escherichia coli cells, activity is highest after 6 hours of cultivation
1.2
-
using 2-phenylpropionitrile as a substrate, at 30°C
1.26
pH 7.5, activity in enzyme preparation after purification, activity of the purified His-tagged protein was 28fold higher than the activity of the wild-type nitrilase
3.26
-
35°C, pH 7.5
4.25
-
20°C, pH 7.5
15.14
initial pH value 8.5
15.39
initial pH value 6.5
16.03
controlled pH value 6.5
16.27
controlled pH value 8.5
18.05
controlled pH value 7.5
18.92
initial pH value 7.5
21.7
Effect of IPG as inducer on the growth and enzyme production by Escherichia coli in a stirred tank reactor
22.8
Effect of lactose as inducer on the growth and enzyme production by Escherichia coli in a stirred tank reactor
90
-
partially purified enzyme
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5
-
about 70% activity at pH 5.0 compared to the pH optimum at 7.0
5
about 70% activity at pH 5.0 compared to the pH optimum at 7.0
6.4 - 9.4
-
about half-maximal activity at pH 6.4 and 9.4, 2-thiopheneacetonitrile as substrate
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
20
-
21% of maximum activity
25 - 65
-
activity range, inactive above
30 - 45
-
more than half maximal activity at 30 and 45°C
70
-
11% of maximum activity
additional information
-
at 50°C and above, rapid loss of activity
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
13000
-
gel filtration
32000
-
14 * 32000
34000
-
10 * 34000, holoenzyme, light scattering analysis
34500
-
10 * 34500, subunit, SDS-PAGE
37700
-
x * 37700, calculated
38000
-
1 * 57000 + 1 * 40000 + 1 * 38000, SDS-PAGE
40000
-
1 * 57000 + 1 * 40000 + 1 * 38000, SDS-PAGE
44000
-
6 * 44000, Alcaligenes faecalis, SDS-PAGE, 6 * 47770, Alcaligenes faecalis, minimal value derived from amino acid composition
44200
-
x * 44200, recombinant enzyme, SDS-PAGE
45000
determined by polyacrylamide gelelectrophorese
47770
-
6 * 44000, Alcaligenes faecalis, SDS-PAGE, 6 * 47770, Alcaligenes faecalis, minimal value derived from amino acid composition
57000
-
1 * 57000 + 1 * 40000 + 1 * 38000, SDS-PAGE
60000
8 * 60000, SDS-PAGE
260000
-
Alcaligenes faecalis, gel filtration on Sepharose 6B
275000
-
Alcaligenes faecalis, gel permeation HPLC
340000
-
holoenzyme, light scattering analysis
412000
-
gel filtration
520000
gel filtration
additional information
-
amino acid composition
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
decamer
-
9 or 10 * 43000, SDS-PAGE
hexamer
homodecamer
homooctamer
homotetradecamer
nonamer
-
9 or 10 * 43000, SDS-PAGE
trimer
-
1 * 57000 + 1 * 40000 + 1 * 38000, SDS-PAGE
additional information
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
2
-
no activity at pH 2
684655
5.5
purified recombinant enzyme, room temperature, 2 h, 10% activity remaining; purified recombinant enzyme, room temperature, 2 h, 10% activity remaining
718670
6.5
purified recombinant enzyme, room temperature, 2 h, 50% activity remaining; purified recombinant enzyme, room temperature, 2 h, 50% activity remaining
718670
6.5 - 9
-
purified recombinant enzyme, room temperature, 2 h, 80-100% activity remaining
718670
6.5
-
half-life 36 min
667345
7 - 10
purified recombinant enzyme, room temperature, 2 h, 90-100% activity remaining; purified recombinant enzyme, room temperature, 2 h, 90-100% activity remaining
718670
7
-
half-life 110 min
667345
7.5
-
half-life 87 min
667345
8
-
half-life 53 min
667345
9.5
-
purified recombinant enzyme, room temperature, 2 h, 65% activity remaining
718670
10
-
purified recombinant enzyme, room temperature, 2 h, 20% activity remaining
718670
10.5
purified recombinant enzyme, room temperature, 2 h, 20% activity remaining; purified recombinant enzyme, room temperature, 2 h, 20% activity remaining
718670
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
15 - 55
exposure of the enzyme at 15, 25, and 35°C for 5 h causes 8%, 15%, and 28% decrease in arylacetonitrilase activity, respectively. A further increase in the temperature to 45 and 55°C results in 58% and 90% decrease in activity, respectively. The half-life of this enzyme at 15, 25, 35, 45, 50, and 55°C is 31 h, 16 h, 9 h, 4 h 30 min, 3 h 20 min, and 2 h 40 min, respectively
25 - 35
purified recombinant enzyme, pH 8.0, 1 h, stable; purified recombinant enzyme, pH 8.0, 1 h, stable
25
-
purified recombinant enzyme, pH 8.0, 1 h, 90% activity remaining
35 - 65
-
purified recombinant enzyme, labile at higher temperature, half-life of 57 min at 40°C, 42 min at 45°C, and 20 min at 55°C, respectively. The activity decreases markedly above 55°C
50 - 60
purified recombinant enzyme, pH 8.0, 1 h, about 10% activity remaining; purified recombinant enzyme, pH 8.0, 1 h, about 10% activity remaining
50 - 70
-
good thermal stability at 50°C, the enzyme completely loses its activity after heat-treatment at 70°C for 30 min
65
-
complete loss of activity
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
immobilisation on poly(ethyleneimine/polyaldehyde)dextran stabilizes
-
in citrate buffer, the activity of the enzyme increases with the increase in the pH (4.0-5.5), whereas in borate and carbonate buffers very little activity is observed
significant stabilization by addition of 1 M ammonium sulfate
-
ORGANIC SOLVENT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
dimethyl sulfoxide
-
2.5 or 5%, up to 20% increase in activity, decrease above
Ethanol
-
5%, 63% enhancement in activity probably due to increased availibaility of substrate. Above 10% ethanol, significant decrease in activity
Methanol
OXIDATION STABILITY
ORGANISM
UNIPROT
LITERATURE
under argon stable for more than 40 h, in oxygen atmosphere 50% loss of activity after 40 h
-
718003
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-20°C, in 10 mM potassium phosphate buffer, pH 7, 10 mM DTT and 50% glycerol, at least 5 months
-
-20°C, pH 7.5, 14 days, 90% loss of activity, addition of 1 M ammonium sulfate almost completely stabilizes
-
-80°C, 0.01 M phosphate, pH 7.5, 1 mM dithiothreitol, 20% glycerol, stable
-
4°C, 0.01 M phosphate, pH 7.5, 1 mM dithiothreitol, rapid loss of activity, presence of 20% glycerol stabilizes
-
4°C, in 10 mM potassium phosphate buffer, pH 7, 10 mM DTT and 50% glycerol, at least 2 months, without glycerol 60% loss of activity within 1 week
-
4°C, in the presence of 60% saturated ammonium sulfate, 2 M NaCl or 30% w/v propanediol, only about 5% loss of activity within 1 week
-
5°C, in 0.02 M citrate buffer, pH 7-8, 10 days, about 50% loss of activity in 0.02 M Tris-HCl buffer, pH 9.3 within 10 days
-
8°C, pH 7.5, 14 days, 75% loss of activity, addition of 1 M ammonium sulfate almost completely stabilizes
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
ammonium sulfate precipitation, DEAE-Sepharose column chromatography, and Sephacryl S-300 gel filtration
cells of bacteria culture are disrupted and centrifugation and the enzyme in the soluble fraction is purified using Ni-NTA affinity chromatography
DEAE Sepharose Fast Flow column chromatography and Sephacryl S-300 gel filtration
-
Ni-NTA column chromatography
-
partial, 259fold
-
recombinant enzyme 2.4fold from Escherichia coli strain BL21-Gold(DE3) to near homogeneity
-
recombinant enzyme 3fold from Escherichia coli strain BL21-Gold(DE3) to near homogeneity; recombinant enzyme 3fold from Escherichia coli strain BL21-Gold(DE3) to near homogeneity
recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography
-
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
DNA and amino acid sequence determination and analysis, sequence comparison, cloning and expression as His-tagged protein in Escherichia coli strains JM109 and BL21(DE3)
-
expressed in Escherichia coli BL21(DE3) cells
-
expressed in Escherichia coli JM109 cells
expressed in Escherichia coli Rosetta2 (DE3) cells
-
expressed in Escherichia coli strain JM109
-
expressed in Pichia pastoris
-
expression in Escherichia coli JM 109, simultanuously with cassava (Manihot esculenta) oxynitrilase
-
expression in Escherichia coli strain BL21-Gold (DE3)
-
expression in Escherichia coli strain BL21-Gold (DE3); expression in Escherichia coli strain BL21-Gold (DE3)
expression under the control of a rhamnose-inducible promoter in Escherichia coli strain JM109
-
recombinant functional expression of the enzyme in the cell envelope of Escherichia coli, where it is accessible for externally added protease
-
The nitrilase gene from Pseudomonas putida is cloned in plasmid pET 21b(+) and over-expressed as histidine-tagged protein in Escherichia coli strain BL21 (DE3)
EXPRESSION
ORGANISM
UNIPROT
LITERATURE
the nitrilase activity from Alcaligenes faecalis ECU0401 increases 4.5fold when the cells are permeabilized with 0.3% (w/v) cetyltrimethylammonium bromide for 20 min at 25°C and pH 6.5
-
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
C163G
-
site-directed mutagenesis
E47Q
-
site-directed mutagenesis
K129I
-
site-directed mutagenesis
C163G
-
site-directed mutagenesis
-
E47Q
-
site-directed mutagenesis
-
K129I
-
site-directed mutagenesis
-
H165N
-
the mutant exhibits a decreased activity for (R,S)-2-phenylpropionitrile compared to the wild type enzyme
W163A
-
the mutant exhibits a decreased activity for (R,S)-2-phenylpropionitrile compared to the wild type enzyme
H165N
-
the mutant exhibits a decreased activity for (R,S)-2-phenylpropionitrile compared to the wild type enzyme
-
W163A
-
the mutant exhibits a decreased activity for (R,S)-2-phenylpropionitrile compared to the wild type enzyme
-
H170N
the mutant exhibits a decreased activity for (R,S)-2-phenylpropionitrile compared to the wild type enzyme
W168A
the mutant forms significantly increases amounts of mandelamide and 2-phenylpropionamide and demonstrates an almost complete inversion of enantioselectivity for the conversion of (R,S)-2-phenylpropionitrile (from R- to S-selectivity)
H170N
-
the mutant exhibits a decreased activity for (R,S)-2-phenylpropionitrile compared to the wild type enzyme
-
W168A
-
the mutant forms significantly increases amounts of mandelamide and 2-phenylpropionamide and demonstrates an almost complete inversion of enantioselectivity for the conversion of (R,S)-2-phenylpropionitrile (from R- to S-selectivity)
-
A114F
-
inactive
A116C
-
the mutant shows activity similar to the wild type enzyme
A116F
-
the mutant shows about 50% of wild type activity
A165E
mutant with very low activities toward (R,S)-mandelonitrile and substrate (R,S)-2-phenylpropionitrile
A165F
the mutant enzyme converts racemic mandelonitrile and (R,S)-2-phenylpropionitrile to increased amounts of the R enantiomers of the corresponding acids
A165G
the mutant forms 4.3% amide and thus produces significantly more amide than the wild type enzyme with the substrate (R,S)-2-phenylpropionitrile
A165H
the mutant enzyme shows a significantly increased relative activity for mandelonitrile (compared to (R,S)-2-phenylpropionitrile)
A165R
mutant with very low activities toward (R,S)-mandelonitrile and substrate (R,S)-2-phenylpropionitrile
A165W
the mutant enzyme converts racemic mandelonitrile and (R,S)-2-phenylpropionitrile to increased amounts of the R enantiomers of the corresponding acids
A165Y
the mutant enzyme converts racemic mandelonitrile and (R,S)-2-phenylpropionitrile to increased amounts of the R enantiomers of the corresponding acids
C163A
-
the mutation results in significantly decreased amounts of amides formed using (R,S)-mandelonitrile and (R,S)-2-phenylpropionitrile as substrates. The mutant demonstrates no significant difference in the enzyme activity compared to the wild type, but shows an extremely low degree of enantioselectivity for the formation of (R)-mandelic acid
C163N
-
the mutation results in significantly increased amounts of amides formed using (R,S)-mandelonitrile and (R,S)-2-phenylpropionitrile as substrates
C163N/A165R
-
the mutant demonstrates increased amide formation capacity in comparison to the mutants carrying only single mutations
C163N/W110I
-
the mutant enzyme forms about 100fold more 2-phenylpropionamide (in relation to the total amount of (R,S)-2-phenylpropionitrile converted) than the wild type, although the relative activity of this mutant for the conversion (R,S)-2-phenylpropionitrile to 2-phenylpropionic acid is only 4% of that observed for the wild type
C163Q
-
the mutation results in significantly increased amounts of amides formed using (R,S)-mandelonitrile and (R,S)-2-phenylpropionitrile as substrates
C163S
-
the mutation results in significantly decreased amounts of amides formed using (R,S)-mandelonitrile and (R,S)-2-phenylpropionitrile as substrates. The mutant demonstrates no significant difference in the enzyme activity compared to the wild type, but shows an extremely low degree of enantioselectivity for the formation of (R)-mandelic acid
C164A
the mutant is devoid of nitrilase activity with the substrate (R,S)-2-phenylpropionitrile
E137A
-
the mutant demonstrates less than 1% of the wild type activity but is still enzymatically competent to convert mandelonitrile to mandelic acid and mandeloamide
G109F
-
inactive
I117F
-
the mutant shows about 50% of wild type activity
L111F
-
the mutant shows about 50% of wild type activity
L113F
-
the mutant shows activity similar to the wild type enzyme
Tyr54C
-
the mutant shows wild type activity but forms significantly decreased relative amounts of atrolactamide from 2-hydroxy-2-phenylpropionitrile
Tyr54P
-
the mutant shows wild type activity but forms significantly decreased relative amounts of atrolactamide from 2-hydroxy-2-phenylpropionitrile
Tyr54V
-
the mutant shows wild type activity but forms significantly decreased relative amounts of atrolactamide from 2-hydroxy-2-phenylpropionitrile
W110F
-
the mutant shows activity similar to the wild type enzyme
Y54A
-
the mutant shows wild type activity but forms significantly decreased relative amounts of atrolactamide from 2-hydroxy-2-phenylpropionitrile
Y54F
-
the mutant shows wild type activity but forms significantly decreased relative amounts of atrolactamide from 2-hydroxy-2-phenylpropionitrile
Y54H
-
the mutant shows wild type activity but forms significantly decreased relative amounts of atrolactamide from 2-hydroxy-2-phenylpropionitrile
Y54L
-
mutant with significantly reduced activity compared to the wild type enzyme
Y54M
-
the mutant shows wild type activity but forms significantly decreased relative amounts of atrolactamide from 2-hydroxy-2-phenylpropionitrile
E137A
-
the mutant demonstrates less than 1% of the wild type activity but is still enzymatically competent to convert mandelonitrile to mandelic acid and mandeloamide
-
Y54A
-
the mutant shows wild type activity but forms significantly decreased relative amounts of atrolactamide from 2-hydroxy-2-phenylpropionitrile
-
Y54F
-
the mutant shows wild type activity but forms significantly decreased relative amounts of atrolactamide from 2-hydroxy-2-phenylpropionitrile
-
Y54H
-
the mutant shows wild type activity but forms significantly decreased relative amounts of atrolactamide from 2-hydroxy-2-phenylpropionitrile
-
Y54I
-
inactive
-
Y54K
-
inactive
-
additional information
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
biotechnology
synthesis