Information on EC 3.5.5.7 - Aliphatic nitrilase

Word Map on EC 3.5.5.7
Please wait a moment until all data is loaded. This message will disappear when all data is loaded.
Specify your search results
Select one or more organisms in this record:
Show additional data
Do not include text mining results
Include (text mining) results (more...)
Include results (AMENDA + additional results, but less precise; more...)


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

EC NUMBER
COMMENTARY hide
3.5.5.7
-
RECOMMENDED NAME
GeneOntology No.
Aliphatic nitrilase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
R-CN + 2 H2O = R-COOH + NH3
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
hydrolysis of C-N bond
hydrolysis of nitriles
-
-
-
-
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
acrylonitrile degradation II
-
-
Microbial metabolism in diverse environments
-
-
Styrene degradation
-
-
degradation of aromatic, nitrogen containing compounds
-
-
SYSTEMATIC NAME
IUBMB Comments
Aliphatic nitrile aminohydrolase
Preferentially hydrolyses aliphatic nitriles, some of which are apparently not substrates for other known nitrilases (EC 3.5.5.1). Substrates include crotononitrile, acrylonitrile and glutaronitrile.
CAS REGISTRY NUMBER
COMMENTARY hide
143236-44-6
Nitrilase (Rhodococcus rhodochrous clone pNK20)
9024-90-2
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
gene nit4, enzyme Nit4
GenBank
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
of teleomorph Pichia guilliermondii NRRL Y-2075, identical sequence of domains D1 and D2; strain UFMG-Y65, isolated from gold mine extraction circuit liquid, inducible enzyme
-
-
Manually annotated by BRENDA team
of teleomorph Pichia guilliermondii NRRL Y-2075, identical sequence of domains D1 and D2; strain UFMG-Y65, isolated from gold mine extraction circuit liquid, inducible enzyme
-
-
Manually annotated by BRENDA team
no activity in Aeropyrum pernix
-
-
-
Manually annotated by BRENDA team
no activity in Pyrococcus furiosus
-
-
-
Manually annotated by BRENDA team
no activity in Pyrococcus horikoshii
-
-
-
Manually annotated by BRENDA team
strain 11387
-
-
Manually annotated by BRENDA team
strain Pf-5
UniProt
Manually annotated by BRENDA team
strain 11388
-
-
Manually annotated by BRENDA team
strain 11388
-
-
Manually annotated by BRENDA team
strain 11401, re-classified from Agrobacterium to Rhizobium
-
-
Manually annotated by BRENDA team
strain 11401, re-classified from Agrobacterium to Rhizobium
-
-
Manually annotated by BRENDA team
strain tg1-A6
-
-
Manually annotated by BRENDA team
strain MTCC 7546
-
-
Manually annotated by BRENDA team
-
Q5N478
UniProt
Manually annotated by BRENDA team
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
(E)-2-methyl-2-butenenitrile + H2O
(E)-2-methyl-2-butenoic acid + NH3
show the reaction diagram
1,4-dicyano-2-butene + H2O
?
show the reaction diagram
-
42.6% of the activity with crotononitrile
-
-
-
1,4-dicyanobutane + H2O
?
show the reaction diagram
114.3% activity compared to 3-cyanopyridine
-
-
?
1,8-dicyanooctane + H2O
?
show the reaction diagram
114.3% activity compared to 3-cyanopyridine
-
-
?
1-cyclohexeneacetonitrile + H2O
1-cyclohexene carboxylate + NH3
show the reaction diagram
-
18.2% of the activity with crotononitrile
-
-
-
1-cyclopenteneacetonitrile + H2O
1-cyclopentene carboxylate + NH3
show the reaction diagram
-
24.9% of the activity with crotononitrile
-
-
-
2,4-dicyano-1-butene + H2O
?
show the reaction diagram
-
127% of the activity with crotononitrile
-
-
-
2,6-dichlorobenzonitrile + H2O
2,6-dichlorobenzamide + ?
show the reaction diagram
2-aminocrotononitrile + H2O
2-aminocrotonic acid + NH3
show the reaction diagram
2-butenenitrile + H2O
2-butenic acid + NH3
show the reaction diagram
-
worst substrate
-
-
?
2-butenenitrile + H2O
2-butenoic acid + NH3
show the reaction diagram
-
-
-
-
?
2-chloroacrylonitrile + H2O
2-chloroacrylic acid + NH3
show the reaction diagram
-
47.1% of the activity with crotononitrile
-
-
-
2-cyanopyridine + H2O
2-pyridine carboxylic acid + NH3
show the reaction diagram
-
-
-
?
2-cyanopyridine + H2O
?
show the reaction diagram
2-furonitrile + H2O
furoic acid + NH3
show the reaction diagram
-
52.0% of the activity with crotononitrile
-
-
-
2-methyl-2-butenenitrile + H2O
2-methyl-2-butenoic acid + NH3
show the reaction diagram
-
14.9% of the activity with crotononitrile
-
-
-
2-methyl-3-butenenitrile + H2O
2-methyl-3-butenoic acid + NH3
show the reaction diagram
-
46.8% of the activity with crotononitrile
-
-
-
2-methylglutaronitrile + H2O
2-methylglutarate + NH3
show the reaction diagram
23.3% activity compared to 3-cyanopyridine
-
-
?
2-methylglutaronitrile + H2O
2-methylglutaric acid + NH3
show the reaction diagram
-
-
-
-
?
2-methylglutaronitrile + H2O
4-cyanopentanoic acid + NH3
show the reaction diagram
77% activity compared to glutaronitrile
-
-
?
2-pentenenitrile + H2O
2-pentenoic acid + NH3
show the reaction diagram
-
6.33% of the activity with crotononitrile
-
-
-
2-thiopheneacetonitrile + H2O
thiophene-2-carboxylate + NH3
show the reaction diagram
-
73.5% of the activity with crotononitrile
-
-
-
2-thiophenecarbonitrile + H2O
2-thiophenecarboxylate + NH3
show the reaction diagram
-
61.1% of the activity with crotononitrile
-
-
-
3,3'-oxydipropionitrile + H2O
? + NH3
show the reaction diagram
-
29.0% of the activity with crotononitrile
-
-
-
3-aminobutyronitrile + H2O
3-aminobutyric acid + NH3
show the reaction diagram
201% activity compared to benzonitrile
-
-
?
3-butenenitrile + H2O
3-butenoic acid + NH3
show the reaction diagram
-
-
-
-
?
3-chloropropionitrile + H2O
3-chloropropanoate + NH3
show the reaction diagram
-
113% of the activity with crotononitrile
-
-
-
3-cyanopyridine + H2O
3-carboxypyridine + NH3
show the reaction diagram
3-cyanopyridine + H2O
?
show the reaction diagram
3-cyanopyridine + H2O
nicotinic acid + NH3
show the reaction diagram
3-ethoxyacrylonitrile + H2O
3-ethoxyacrylic acid + NH3
show the reaction diagram
3-ethoxybenzonitrile + H2O
3-ethoxybenzoate + NH3
show the reaction diagram
-
21.1% of the activity with crotononitrile
-
-
-
3-methoxybenzonitrile
3-methoxybenzoate + NH3
show the reaction diagram
-
24.8% of the activity with crotononitrile
-
-
-
3-nitrobenzonitrile + H2O
3-nitrobenzoate + NH3
show the reaction diagram
-
74.5% of the activity with crotononitrile
-
-
-
3-nitrobenzonitrile + H2O
3-nitrobenzoic acid + NH3
show the reaction diagram
110% activity compared to benzonitrile
-
-
?
3-pentenenitrile + H2O
3-pentenoic acid + NH3
show the reaction diagram
-
132% of the activity with crotononitrile
-
-
-
3-phenylpropionitrile + H2O
3-phenylpropanoate + NH3
show the reaction diagram
147.3% activity compared to 3-cyanopyridine
-
-
?
3-phenylpropionitrile + H2O
3-phenylpropionic acid + NH3
show the reaction diagram
3-phenylpropionitrile + H2O
?
show the reaction diagram
59% activity compared to glutaronitrile
-
-
?
3-thiopheneacetonitrile + H2O
thiophene-3-carboxylate + NH3
show the reaction diagram
-
66.3% of the activity with crotononitrile
-
-
-
3-tolunitrile + H2O
3-methylbenzenecarboxylic acid + NH3
show the reaction diagram
-
67.9% of the activity with crotononitrile
-
-
-
3-xylylene dicyanide + H2O
?
show the reaction diagram
-
14.62% of the activity with crotononitrile
-
-
-
4-aminobutyronitrile + H2O
4-aminobutyric acid + NH3
show the reaction diagram
251% activity compared to benzonitrile
-
-
?
4-chlorobutyronitrile + H2O
4-chlorobutyrate + NH3
show the reaction diagram
4-cyano-1-cyclohexene + H2O
1-cyclohexene-4-carboxylate + NH3
show the reaction diagram
-
7.03% of the activity with crotononitrile
-
-
-
4-cyanopyridine + H2O
4-carboxypyridine + NH3
show the reaction diagram
-
9.71% of the activity with crotononitrile
-
-
-
4-cyanopyridine + H2O
4-pyridinecarboxylic acid + NH3
show the reaction diagram
4-cyanopyridine + H2O
?
show the reaction diagram
4-nitrobenzonitrile + H2O
4-nitrobenzoic acid + NH3
show the reaction diagram
151% activity compared to benzonitrile
-
-
?
acetamide + H2O
?
show the reaction diagram
acetonitrile + H2O
?
show the reaction diagram
5% activity compared to glutaronitrile
-
-
?
acetonitrile + H2O
acetate + NH3
show the reaction diagram
acetonitrile + H2O
acetic acid + NH3
show the reaction diagram
-
-
-
-
?
acrylamide + H2O
?
show the reaction diagram
acrylonitrile + H2O
acrylate + NH3
show the reaction diagram
acrylonitrile + H2O
acrylic acid + NH3
show the reaction diagram
adipamide + H2O
?
show the reaction diagram
-
-
-
-
?
adiponitrile + H2O
adipate + NH3
show the reaction diagram
adiponitrile + H2O
adipic acid + NH3
show the reaction diagram
aminoacetonitrile + H2O
aminoacetic acid + NH3
show the reaction diagram
benzamide + H2O
?
show the reaction diagram
-
-
-
-
?
benzonitrile + H2O
benzoate + NH3
show the reaction diagram
benzonitrile + H2O
benzoic acid + NH3
show the reaction diagram
beta-cyano-L-alanine + H2O
asparagine + aspartic acid
show the reaction diagram
-
60:40 mixture
-
?
butyronitrile + H2O
?
show the reaction diagram
6% activity compared to glutaronitrile
-
-
?
butyronitrile + H2O
butyrate + NH3
show the reaction diagram
butyronitrile + H2O
butyric acid + NH3
show the reaction diagram
capronitrile + H2O
hexanoate + NH3
show the reaction diagram
-
38.7% of the activity with crotononitrile
-
-
-
chloroacetonitrile + H2O
chloroacetate + NH3
show the reaction diagram
-
61.7% of the activity with crotononitrile
-
-
-
cinnamonitrile + H2O
cinnamate + NH3
show the reaction diagram
cis-crotononitrile + H2O
crotonic acid + NH3
show the reaction diagram
crotonitrile + H2O
?
show the reaction diagram
7% activity compared to glutaronitrile
-
-
?
crotonitrile + H2O
crotonic acid + NH3
show the reaction diagram
cyano-5-valeramide + H2O
glutaric acid 5-amide + NH3
show the reaction diagram
-
at 28% of the activity with adiponitrile
-
-
-
cyano-5-valeric acid + H2O + H2O
?
show the reaction diagram
-
at 28% of the activity with adiponitrile
-
-
-
cyanoacetic acid ethyl ester + H2O
acetic acid ethyl ester + NH3
show the reaction diagram
-
117% of the activity withcrotononitrile
-
-
-
cyanopyrazine + H2O
pyrazine-2-carboxylate + NH3
show the reaction diagram
-
17.3% of the activity with crotononitrile
-
-
-
cyclopentanecarbonitrile + H2O
cyclopentanoic acid + NH3
show the reaction diagram
-
11.7% of the activity with crotononitrile
-
-
-
cyclopentanocarbonitrile + H2O
cyclopentanocarbonate + NH3
show the reaction diagram
-
-
-
-
?
cyclopropanecarbonitrile + H2O
cyclopropanoic acid + NH3
show the reaction diagram
-
22.9% of the activity with crotononitrile
-
-
-
diaminomaleonitrile + H2O
diaminomaleic acid + NH3
show the reaction diagram
-
18.6% of the activity with crotononitrile
-
-
-
dimethylmalononitrile + H2O
dimethylmalonic acid + NH3
show the reaction diagram
-
-
-
-
?
diphenylacetonitrile + H2O
diphenylacetic acid + NH3
show the reaction diagram
72% activity compared to benzonitrile
-
-
?
dodecanenitrile + H2O
dodecanoate + NH3
show the reaction diagram
13.5% activity compared to 3-cyanopyridine
-
-
?
fumaronitrile + H2O
fumarate + NH3
show the reaction diagram
fumaronitrile + H2O
fumaric acid + NH3
show the reaction diagram
glutaramide + H2O
?
show the reaction diagram
-
-
-
-
?
glutaronitrile + H2O
glutarate + NH3
show the reaction diagram
glutaronitrile + H2O
glutaric acid + NH3
show the reaction diagram
glycolonitrile + 2 H2O
glycolic acid + NH3
show the reaction diagram
glycolonitrile + H2O
ammonium glycolate + ?
show the reaction diagram
heptanenitrile + H2O
heptanoic acid + NH3
show the reaction diagram
-
-
-
-
?
hexanenitrile + H2O
hexanoic acid + NH3
show the reaction diagram
-
-
-
-
?
hydrocinnamonitrile + H2O
hydrocinnamic acid + NH3
show the reaction diagram
-
-
-
-
?
iminodiacetonitrile + H2O
? + NH3
show the reaction diagram
isobutyramide + H2O
?
show the reaction diagram
-
-
-
-
?
isobutyronitrile + H2O
isobutyrate + NH3
show the reaction diagram
isobutyronitrile + H2O
isobutyric acid + NH3
show the reaction diagram
-
-
-
-
?
isocapronitrile + H2O
isohexanoate + NH3
show the reaction diagram
-
41.5% of the activity with crotononitrile
-
-
-
isophthalonitrile + H2O
isophthalate + NH3
show the reaction diagram
-
66.1% of the activity with crotononitrile
-
-
-
isovaleronitrile + H2O
isovaleric acid + NH3
show the reaction diagram
-
-
-
-
?
m-tolunitrile + H2O
m-methylbenzoate + NH3
show the reaction diagram
-
-
-
?
malonitrile + H2O
malonic acid + NH3
show the reaction diagram
-
-
-
-
?
malononitrile + H2O
malonic acid + NH3
show the reaction diagram
mandelonitrile + 2 H2O
mandelic acid + NH3
show the reaction diagram
-
-
-
-
?
mandelonitrile + H2O
mandelic acid + NH3
show the reaction diagram
69% activity compared to benzonitrile
-
-
?
metacrylonitrile + H2O
metacrylic acid + NH3
show the reaction diagram
-
-
-
-
?
methacrylonitrile + H2O
methacrylic acid + NH3
show the reaction diagram
methoxyacetonitrile + H2O
methoxyacetate + NH3
show the reaction diagram
-
37.4% of the activity with crotononitrile
-
-
-
N-butyronitrile + 2 H2O
N-butyric acid + NH3
show the reaction diagram
N-methyl-beta-alaninenitrile + H2O
N-methyl-beta-Ala + NH3
show the reaction diagram
-
11.4% of the activity with crotononitrile
-
-
-
pentanenitrile + H2O
pentanoate + NH3
show the reaction diagram
62% activity compared to 3-cyanopyridine
-
-
?
phenylacetonitrile + H2O
phenylacetate + NH3
show the reaction diagram
-
27.3% of the activity with crotononitrile
-
-
-
phenylacetonitrile + H2O
phenylacetic acid + NH3
show the reaction diagram
-
-
-
-
?
pimelonitrile + H2O
pimelic acid + NH3
show the reaction diagram
-
27.3% of the activity with crotononitrile
-
-
-
piperonylonitrile + H2O
(3,4-methylenedioxy)benzoate + NH3
show the reaction diagram
-
6.8% of the activity with crotononitrile
-
-
-
potassium cyanide + H2O
?
show the reaction diagram
-
-
-
-
?
propionitrile + H2O
?
show the reaction diagram
5% activity compared to glutaronitrile
-
-
?
propionitrile + H2O
propionate + NH3
show the reaction diagram
propionitrile + H2O
propionic acid + NH3
show the reaction diagram
R-CN + H2O
R-COOH + NH3
show the reaction diagram
-
enzyme uses both aliphatic and aromatic nitriles, no formation of amide intermediate
-
-
?
racemic Ibu-CN + H2O
fumaric acid + NH3
show the reaction diagram
sebaconitrile + H2O
? + NH3
show the reaction diagram
sebaconitrile + H2O
sebaconic acid + NH3
show the reaction diagram
-
-
-
-
?
suberonitrile + H2O
decanedioic acid + NH3
show the reaction diagram
-
21.4% of the activity with crotononitrile
-
-
-
succinamide + H2O
?
show the reaction diagram
-
-
-
-
?
succinodinitrile + H2O
?
show the reaction diagram
-
-
-
-
?
succinonitrile + H2O
succinate + NH3
show the reaction diagram
succinonitrile + H2O
succinic acid + NH3
show the reaction diagram
thiophene-3-carbonitrile + H2O
?
show the reaction diagram
51.2% activity compared to 3-cyanopyridine
-
-
?
trans-crotononitrile + H2O
crotonic acid + NH3
show the reaction diagram
valeronitrile + H2O
pentanoate + NH3
show the reaction diagram
-
40.8% of the activity with crotononitrile
-
-
-
valeronitrile + H2O
valeric acid + NH3
show the reaction diagram
-
-
-
-
?
additional information
?
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Ca2+
about 115% activity at 1 mM
Fe2+
about 110% activity at 1 mM
Fe3+
about 125% activity at 1 mM
Mg2+
about 125% activity at 1 mM
Mn2+
about 120% activity at 1 mM
Zn2+
about 120% activity at 1 mM
additional information
-
enzyme contains no metal
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2-mercaptoethanol
-
5 mM, 29% inhibition
5,5'-dithiobis(2-nitrobenzoate)
-
complete inhibition at 5 mM
acetone
-
10%, complete inhibition; complete inhibition at 10% acetone
Ba2+
about 55% residual activity at 1 mM
Cr3+
-
5 mM, 40% inhibition; 60% residual activity at 5 mM
Dichloromethane
about 30% residual activity at 5% (v/v)
dithiothreitol
-
5 mM, 50% inhibition
DTNB
-
5 mM, complete inhibition
ethanol
ethylene glycol
-
80% inhibition at 50%; 80% inhibition at 50% ethylene glycol
hexane
about 20% residual activity at 5% (v/v)
HgCl2
-
inhibition partially reversed by DTT, 2-mercaptoethanol, or reduced glutathione
iodacetamide
-
complete inhibition at 5 mM
iodacetate
-
complete inhibition at 5 mM
iodoacetamide
-
5 mM, complete inhibition
iodoacetate
-
5 mM, complete inhibition
L-Cys
-
5 mM, 27% inhibition
Li+
about 80% residual activity at 1 mM
Malononitrile
-
concentrations above 12 mM have a strong inhibitory effect; strong inhibition at 12 mM
methanol
Mg2+
93% residual activity at 5 mM
N-ethylmaleimide
0.1 mM, 48% residual activity
NH3
-
inhibition partially reversed by DTT, 2-mercaptoethanol, or reduced glutathione
p-chloromercuribenzoate
-
complete inhibition at 1 mM
p-hydroxymercuribenzoate
-
1 mM, complete inhibition; complete inhibition at 1 mM
phenyl hydrazine
-
53% residual activity at 5 mM; 5 mM, 47% inhibition
Propanol
about 22% residual activity at 5% (v/v)
pyridoxal 5'-phosphate
0.1 mM, 30% residual activity
Toluene
about 5% residual activity at 5% (v/v)
additional information
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2-mercaptoethanol
-
activates at 1 mM
Benzonitrile
dithiothreitol
-
1 mM, 23% inhibition; activates at 1 mM
EDTA
about 130% activity at 1 mM
ethylene glycol
-
20% increase of activity at 10% ethylene glycol
L-cysteine
-
activates at 1 mM
additional information
-
there is no appreciable variation in the activity of enzyme at any concentration of glycerol
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
7.95 - 9.16
2-butenenitrile
0.82 - 5
2-Cyanopyridine
3.16 - 73.1
3-Cyanopyridine
11.26
acetonitrile
-
pH 7.0, 37C, partially purified enzyme
1.14 - 23.3
acrylonitrile
0.688 - 10.28
adiponitrile
0.49 - 23.8
Benzonitrile
18.9
Crotonitrile
-
-
2 - 11.45
Fumaronitrile
1.15 - 47
glutaronitrile
0.47 - 1.85
m-tolunitrile
3.47
Malononitrile
-
in 20 mM potassium phosphate, pH 7.4, with 1 mM dithiothreitol, at 80C; pH 7.4, 80C
0.75
potassium cyanide
-
pH 7.0, 37C, partially purified enzyme
43.9
Propionitrile
pH 7.0, 30C
25.3
sebaconitrile
pH 7.0, 30C
17.9
Succinonitrile
pH 7.0, 30C
additional information
additional information
-
kinetics
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.03 - 0.19
2-butenenitrile
0.00012 - 2.05
2-Cyanopyridine
0.45 - 1.21
3-Cyanopyridine
0.33
acrylonitrile
Pseudomonas fluorescens
Q4KCL8
pH 7.0, 30C
0.00018 - 0.0038
adiponitrile
0.00014 - 0.98
Benzonitrile
7.42 - 15.13
Fumaronitrile
0.00018 - 5
glutaronitrile
0.0001 - 0.00038
m-tolunitrile
0.17
Propionitrile
Pseudomonas fluorescens
Q4KCL8
pH 7.0, 30C
3.3
sebaconitrile
Pseudomonas fluorescens
Q4KCL8
pH 7.0, 30C
8.8
Succinonitrile
Pseudomonas fluorescens
Q4KCL8
pH 7.0, 30C
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.00328 - 0.0239
2-butenenitrile
144409
1.224 - 1.814
2-Cyanopyridine
1991
0.141 - 0.357
3-Cyanopyridine
1784
0.531 - 0.806
Benzonitrile
1135
0.689 - 1.679
Fumaronitrile
16086
0.483
glutaronitrile
uncultured bacterium
E1A0Z9
at 30C in 50 mM sodium phosphate buffer (pH 7.5) and 1 mM dithiothreitol
3634
IC50 VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
150 - 2200
additional information
Meyerozyma guilliermondii
-
minimal inhibition concentration MIC values of nitrile and amide substrates, the values lay between 150 mM and 2200 mM, overview
-
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.00595
-
recombinant enzyme from crude extract
0.05
-
mutant enzyme W146F, with 2-butenenitrile as substrate, at pH 7.0 and 30C
0.1419
-
recombinant enzyme after 23.85fold purification
0.29
-
wild type enzyme, with 2-butenenitrile as substrate, at pH 7.0 and 30C
0.68
-
wild type enzyme, with 3-cyanopyridine as substrate, at pH 7.0 and 30C
0.97
-
wild type enzyme, with benzonitrile as substrate, at pH 7.0 and 30C
1
-
mutant enzyme W146F, with benzonitrile as substrate, at pH 7.0 and 30C
1.18
-
mutant enzyme W146Y, with benzonitrile as substrate, at pH 7.0 and 30C
1.4
mutant enzyme Y175A, using succinonitrile as a substrate
1.47
-
mutant enzyme W146H, with benzonitrile as substrate, at pH 7.0 and 30C
1.57
-
mutant enzyme W146Y, with fumaronitrile as substrate, at pH 7.0 and 30C
1.6
-
mutant enzyme W146F, with 3-cyanopyridine as substrate, at pH 7.0 and 30C
1.69
-
mutant enzyme W146Y, with 3-cyanopyridine as substrate, at pH 7.0 and 30C
1.8
mutant enzyme K9A, using sebacconitrile as a substrate
1.82
-
mutant enzyme W146H, with 3-cyanopyridine as substrate, at pH 7.0 and 30C
1.85
-
mutant enzyme W146F, with fumaronitrile as substrate, at pH 7.0 and 30C
1.9
substrate fumaronitrile
1.95
-
mutant enzyme W146H, with fumaronitrile as substrate, at pH 7.0 and 30C
2.5
-
crude enzyme extract, substrate 2-cyanopyridine
2.72
-
mutant enzyme W146H, with 2-cyanopyridine as substrate, at pH 7.0 and 30C
2.93
-
mutant enzyme W146F, with 2-cyanopyridine as substrate, at pH 7.0 and 30C
3.08
-
mutant enzyme W146Y, with 2-cyanopyridine as substrate, at pH 7.0 and 30C
3.2
-
wild type enzyme, with fumaronitrile as substrate, at pH 7.0 and 30C
3.5
-
crude enzyme extract, substrate adipamide
3.6
-
crude enzyme extract, substrate glutaronitrile
4.3
wild type enzyme, using succinonitrile as a substrate
4.6
-
crude enzyme extract, substrate adiponitrile
4.8
mutant enzyme L79A, using sebacconitrile as a substrate
5
-
crude enzyme extract, substrate benzamide
5.6
-
crude enzyme extract, substrate propionitrile
5.7
mutant enzyme Y175A, using adiponitrile as a substrate
6.8
-
crude enzyme extract, substrate cyclopentanocaronitrile
6.9
-
crude enzyme extract, substrate benzonitrile
7.2
-
crude enzyme extract, substrate 3-cyanopyridine
7.3
-
crude enzyme extract, substrate butyronitrile
8.1
-
crude enzyme extract, substrate acetamide
8.3
-
crude enzyme extract, substrate succinamide
8.9
-
crude enzyme extract, substrate acrylamide
9.2
-
crude enzyme extract, substrate isobutyramide; crude enzyme extract, substrate isobutyronitrile
9.7
-
crude enzyme extract, substrate metacrylonitrile
10.4
-
crude enzyme extract, substrate acrylonitrile
13.5
-
crude enzyme extract, substrate acetonitrile
15.8
-
crude enzyme extract, substrate succinonitrile
16.1
wild type enzyme, using glutaronitrile as a substrate
17.53
-
partially purified enzyme
17.7
mutant enzyme K9A, using glutaronitrile as a substrate
22.7
mutant enzyme L79A, using glutaronitrile as a substrate
29.2
mutant enzyme L79A, using adiponitrile as a substrate
31.4
wild type enzyme, using adiponitrile as a substrate
40.8
mutant enzyme K9A, using adiponitrile as a substrate
additional information
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5 - 9
Nit1 tolerates a pH range between 5 and 9 with an activity of more than 50%
6.6 - 7.4
-
-
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
25 - 45
more than 60% activity between 25 and 45C
30 - 50
about 55% activity at 30C, about 80% activity at 35C, 100% activity at 40C, about 70% activity at 45C, and about 30% activity at 50C
37 - 50
-
-
40 - 55
-
40C: about 40% of maximal activity, 55C: about 60% of maximal activity, crotonitrile
pI VALUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5
calculated from amino acid sequence
5.1
-
calculated from amino acid sequence
5.2
-
calculated from amino acid sequence
5.3
-
isoelectric focusing
5.4
isoelectric focusing and calculated
5.7
-
calculated from amino acid sequence
5.9
calculated from amino acid sequence
6
Q5N478
calculated from amino acid sequence
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
additional information
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
30000
-
2 * 30000, SDS-PAGE
33000
4 * 33000, SDS-PAGE
36320
-
calculated from amino acid sequence
36350
Q5N478
calculated from amino acid sequence
36550
calculated from amino acid sequence
37000
x * 37000, SDS-PAGE
37970
calculated from amino acid sequence
40190
-
calculated from amino acid sequence
42275
-
x * 42275, calculation from nucleotide sequence
42280
-
calculated from amino acid sequence
43000
-
14 * 41000 or 14 * 43000
59800
-
gel filtration
138000
gel filtration
560000
gel filtration
604000
-
equilibrium sedimentation
650000
700000
gel filtration
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
dimer
-
2 * 30000, SDS-PAGE
hexadecamer
homodecamer
-
10 * 40000
homodimer
homotetradecamer
homotetradecamer or homohexacecamer
monomer
pentadecamer
tetramer
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
molecular modeling of enzyme and docking studies using aromatic and aliphatic nitrile substrates
vapor diffusion method, using 15% (w/v) PEG 5000MME, 1 mM dithiothreitol, 3% tascimate and 100 mM HEPES, pH 6.8
-
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6 - 8
-
25C, 45 min, less than 30% loss of activity
32506
6.6 - 7
-
enzymatic activity increases in the pH range 6.6 to 7.0 and then starts decreasing
687941
10
-
25C, 45 min, 40% loss of activity
32506
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
16
50% of initial activity after 80 h
25
-
pH 7.5, 1 h, stable
35
-
pH 7.5, 1 h, 8% loss of activity
40
-
pH 7.5, 1 h, 13% loss of activity
40 - 60
up to 40C Nit1 is stable for 1 h, while the loss of activity is about 50% after 30 min at 50C, the enzyme is completely inactivated after 10 min incubation at 60C
50
-
pH 7.5, 1 h, 93% loss of activity
65
50% of initial activity after 45 min
70 - 90
-
the recombinant enzyme is highly thermostable with a half-life of 25 h at 70C, 9 h at 80C, and 6 h at 90C
70
-
25 h, recombinant enzyme loses about 50% of maximal activity
80
-
half-life: 9 h
90
-
half-life: 6 h
113
-
Tm-value: 112.7C
120
-
irreversible denaturation after heating to 120C
ORGANIC SOLVENT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Methanol
activity assay in presence of 10% v/v
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
3-step procedure, partially, 10.9fold
-
; Q-Sepharose Fast Flow column chromatography and Sephadex PD-10 column gel filtration
-
ammonium sulfate precipitation and gel filtration
-
HisTrap HP 9 column chromatography
Ni-NTA column chromatography
Ni-NTA column chromatography and Superdex 200 gel filtration
Ni-NTA column chromatography, and gel filtration
Ni2+ affinity column chromatography and Superdex 75 gel filtration
-
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
DNA sequence determination and analysis
-
DNA sequence determination and analysis, overexpression in Escherichia coli
-
expressed in Escherichia coli
-
expressed in Escherichia coli BL21 (DE3) cells
expressed in Escherichia coli BL21(DE3) cells
expressed in Escherichia coli Rosetta (DE3) cells
expressed in Escherichia coli strain ER2566
expressed in Escherichia coli; expression in Escherichia coli
-
expression in Escherichia coli
overproduction in Escherichia coli
-
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
F168V/L201N
-
the mutant shows increased activity and specificity compared to the wild type enzyme
L201Q
-
the mutant shows increased activity and specificity compared to the wild type enzyme
T210G
-
inactive
F168V/L201N
-
the mutant shows increased activity and specificity compared to the wild type enzyme
-
L201Q
-
the mutant shows increased activity and specificity compared to the wild type enzyme
-
T210G
-
inactive
-
C170A
-
mutant enzymes Cys170Ala and Cys170Ser show no catalytic activity
C170S
-
mutant enzymes Cys170Ala and Cys170Ser show no catalytic activity
K9A
activity similar to wild-type; increased specific activities
L79A
increased specific activities; slight decrease in activity towards aromatic nitriles
R129A
inactive; loss of enzymatic activiy towards all nitriles
Y142A
loss of activity towards aliphatic nitriles; mutant exhibits slightly higher kcat/Km values for aromatic nitriles and shows no activity toward aliphatic nitriles; mutant specific for aromatic nitrile substrates
Y142D
complete loss of activity
Y142E
complete loss of activity
Y142F
mutant shows slightly lower kcat/Km values compared to the wild type enzyme; substrate specificity similar to wild-type
Y142G
complete loss of activity
Y142I
low specific activities; very low activity
Y142L
low specific activities; mutant specific for aromatic nitrile substrates
Y142N
complete loss of activity
Y142R
complete loss of activity
Y142S
mutant exhibits slightly higher kcat/Km values for aromatic nitriles and shows no activity toward aliphatic nitriles; mutant specific for aromatic nitrile substrates
Y142V
mutant specific for aromatic nitrile substrates
Y175A
activity below 6 micromol/min mg; increased specific activities
C170A
-
mutant enzymes Cys170Ala and Cys170Ser show no catalytic activity
-
C170S
-
mutant enzymes Cys170Ala and Cys170Ser show no catalytic activity
-
C169A
-
inactive
E142A
-
inactive
E53A
-
inactive
K135A
-
inactive
N118A
-
inactive
W146A
-
inactive
W146C
-
inactive
W146D
-
inactive
W146E
-
inactive
W146F
-
the mutant shows reduced activity compared to the wild type enzyme
W146G
-
inactive
W146H
-
the mutant shows reduced activity compared to the wild type enzyme
W146I
-
inactive
W146K
-
inactive
W146L
-
inactive
W146M
-
inactive
W146N
-
inactive
W146P
-
inactive
W146Q
-
inactive
W146R
-
inactive
W146S
-
inactive
W146T
-
inactive
W146V
-
inactive
W146Y
-
the mutant shows reduced activity compared to the wild type enzyme
Y59A
-
inactive
additional information
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
biotechnology
environmental protection
industry
synthesis
Show AA Sequence (374 entries)
Please use the Sequence Search for a specific query.