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show all sequences of 3.5.5.5

A novel nitrilase, arylacetonitrilase, of Alcaligenes faecalis JM3. Purification and characterization

Nagasawa, T.; Mauger, J.; Yamada, H.; Eur. J. Biochem. 194, 765-772 (1990)

Data extracted from this reference:

Activating Compound
Activating Compound
Commentary
Organism
Structure
2-mercaptoethanol
activation
Alcaligenes faecalis
dithiothreitol
activation
Alcaligenes faecalis
Sodium hydrosulfide
activation
Alcaligenes faecalis
Inhibitors
Inhibitors
Commentary
Organism
Structure
2,2'-dipyridyl
weak
Alcaligenes faecalis
3-Methyl-2-benzothiazolinone hydrazone
weak
Alcaligenes faecalis
8-hydroxyquinoline
weak
Alcaligenes faecalis
Ag2+
strong
Alcaligenes faecalis
ascorbic acid
-
Alcaligenes faecalis
Cd2+
-
Alcaligenes faecalis
Co2+
-
Alcaligenes faecalis
Cu2+
strong
Alcaligenes faecalis
D-cycloserine
weak
Alcaligenes faecalis
diethyldithiocarbamate
-
Alcaligenes faecalis
Disodium 4,5-dihydroxy-m-benzene-disulfonate
weak
Alcaligenes faecalis
DTNB
strong
Alcaligenes faecalis
H2O2
-
Alcaligenes faecalis
Hg2+
strong
Alcaligenes faecalis
hydroxylamine
weak
Alcaligenes faecalis
iodoacetate
weak
Alcaligenes faecalis
KCN
weak
Alcaligenes faecalis
L-cysteine
-
Alcaligenes faecalis
additional information
little or no inhibition by Li+, Na+, Mg2+, Ca2+, Ba2+, Mn2+, Fe2+, Pb2+, Fe3+, Al3+, NaN3, NaF, cysteamine, aminoguanidine, L/D-penicillamine, 1,10-phenanthroline or EDTA
Alcaligenes faecalis
N-ethylmaleimide
-
Alcaligenes faecalis
Ni2+
-
Alcaligenes faecalis
p-chloromercuribenzoate
strong
Alcaligenes faecalis
phenylhydrazine
-
Alcaligenes faecalis
Semicarbazide
weak
Alcaligenes faecalis
Sn2+
-
Alcaligenes faecalis
Urea
weak
Alcaligenes faecalis
Zn2+
-
Alcaligenes faecalis
KM Value [mM]
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.009
-
3-Indoleacetonitrile
-
Alcaligenes faecalis
0.01
-
benzylcyanide
-
Alcaligenes faecalis
0.015
-
2-Chlorobenzylcyanide
-
Alcaligenes faecalis
0.016
-
3-Thiopheneacetonitrile
-
Alcaligenes faecalis
0.022
-
4-Aminobenzylcyanide
-
Alcaligenes faecalis
0.023
-
3-tolylacetonitrile
-
Alcaligenes faecalis
0.065
-
3-Chlorobenzylcyanide
-
Alcaligenes faecalis
0.091
-
2-Thiopheneacetonitrile
-
Alcaligenes faecalis
0.105
-
4-Nitrobenzylcyanide
-
Alcaligenes faecalis
0.123
-
4-tolylacetonitrile
-
Alcaligenes faecalis
0.13
-
4-Fluorobenzylcyanide
-
Alcaligenes faecalis
0.333
-
4-Chlorobenzylcyanide
-
Alcaligenes faecalis
0.66
-
acrylonitrile
-
Alcaligenes faecalis
Molecular Weight [Da]
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
additional information
-
amino acid composition
Alcaligenes faecalis
44000
-
6 * 44000, Alcaligenes faecalis, SDS-PAGE, 6 * 47770, Alcaligenes faecalis, minimal value derived from amino acid composition
Alcaligenes faecalis
47770
-
6 * 44000, Alcaligenes faecalis, SDS-PAGE, 6 * 47770, Alcaligenes faecalis, minimal value derived from amino acid composition
Alcaligenes faecalis
260000
-
Alcaligenes faecalis, gel filtration on Sepharose 6B
Alcaligenes faecalis
275000
-
Alcaligenes faecalis, gel permeation HPLC
Alcaligenes faecalis
Natural Substrates/ Products (Substrates)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
Arylacetonitrile + H2O
Alcaligenes faecalis
pathway in nitrile catabolism, inducible enzyme
?
-
-
-
Organic Solvent Stability
Organic Solvent
Commentary
Organism
Methanol
stable in the presence of 10% w/v methanol
Alcaligenes faecalis
Organism
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
Alcaligenes faecalis
-
JM3
-
Alcaligenes faecalis JM3
-
JM3
-
Purification (Commentary)
Commentary
Organism
-
Alcaligenes faecalis
Specific Activity [micromol/min/mg]
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
144
-
-
Alcaligenes faecalis
Storage Stability
Storage Stability
Organism
-20°C, in 10 mM potassium phosphate buffer, pH 7, 10 mM DTT and 50% glycerol, at least 5 months
Alcaligenes faecalis
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
Alcaligenes faecalis
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
Alcaligenes faecalis
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
Alcaligenes faecalis
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
1-cyclohexenylacetonitrile + H2O
at 3.3% the rate of 4-chlorobenzylcyanide hydrolysis
32503
Alcaligenes faecalis
1-cyclohexenylacetate + NH3
-
-
-
-
2,4-dichlorobenzylcyanide + H2O
at 6.5% the rate of 4-chlorobenzylcyanide hydrolysis
32503
Alcaligenes faecalis
2,4-dichlorophenylacetic acid + NH3
-
-
-
-
2-chlorobenzylcyanide + H2O
at 6% the rate of 4-chlorobenzylcyanide hydrolysis
32503
Alcaligenes faecalis
2-chlorophenylacetic acid + NH3
-
-
-
-
2-methoxybenzylcyanide + H2O
at 2% the rate of 4-chlorobenzylcyanide hydrolysis
32503
Alcaligenes faecalis
2-methoxyphenylacetic acid + NH3
-
-
-
-
2-thiopheneacetonitrile + H2O
ir, at 90% the rate of 4-chlorobenzylcyanide hydrolysis
32503
Alcaligenes faecalis
2-thiopheneacetic acid + NH3
not via thiopheneacetamide
32503
Alcaligenes faecalis
-
3,4-(methylenedioxy)phenylacetonitrile + H2O
at 65% the rate of 4-chlorobenzylcyanide hydrolysis
32503
Alcaligenes faecalis
3,4-(methylendioxy)phenylacetic acid + NH3
-
-
-
-
3,4-dichlorobenzylcyanide + H2O
at 14% the rate of 4-chlorobenzylcyanide hydrolysis
32503
Alcaligenes faecalis
3,4-dichlorophenylacetic acid + NH3
-
-
-
-
3-chlorobenzylcyanide + H2O
at 43% the rate of 4-chlorobenzylcyanide hydrolysis
32503
Alcaligenes faecalis
3-chlorophenylacetic acid + NH3
-
-
-
-
3-indoleacetonitrile + H2O
at 9% the rate of 4-chlorobenzylcyanide hydrolysis
32503
Alcaligenes faecalis
3-indoleacetic acid + NH3
-
32503
Alcaligenes faecalis
-
3-indoleacetonitrile + H2O
at 9% the rate of 4-chlorobenzylcyanide hydrolysis
32503
Alcaligenes faecalis JM3
3-indoleacetic acid + NH3
-
32503
Alcaligenes faecalis JM3
-
3-phenylpropionitrile + H2O
at 26% the rate of 4-chlorobenzylcyanide hydrolysis
32503
Alcaligenes faecalis
3-phenylpropionic acid + NH3
-
-
-
-
3-pyridineacetonitrile + H2O
at 77% the rate of 4-chlorobenzylcyanide hydrolysis
32503
Alcaligenes faecalis
?
-
-
-
-
3-thiopheneacetonitrile + H2O
at 24% the rate of 4-chlorobenzylcyanide hydrolysis
32503
Alcaligenes faecalis
3-thiopheneacetic acid + NH3
-
32503
Alcaligenes faecalis
-
4-aminobenzylcyanide + H2O
at 44% the rate of 4-chlorobenzylcyanide hydrolysis
32503
Alcaligenes faecalis
4-aminophenylacetic acid + NH3
-
-
-
-
4-bromobenzylcyanide + H2O
at 26% the rate of 4-chlorobenzylcyanide hydrolysis
32503
Alcaligenes faecalis
?
-
-
-
-
4-bromobenzylcyanide + H2O
at 26% the rate of 4-chlorobenzylcyanide hydrolysis
32503
Alcaligenes faecalis JM3
?
-
-
-
-
4-chlorobenzylcyanide + H2O
i.e. p-chlorophenylacetonitrile, best substrate
32503
Alcaligenes faecalis
4-chlorophenylacetic acid + NH3
-
32503
Alcaligenes faecalis
-
4-chlorobenzylcyanide + H2O
i.e. p-chlorophenylacetonitrile, best substrate
32503
Alcaligenes faecalis JM3
4-chlorophenylacetic acid + NH3
-
32503
Alcaligenes faecalis JM3
-
4-fluorobenzylcyanide + H2O
i.e. p-fluorophenylacetonitrile, at 96% the rate of 4-chlorobenzylcyanide hydrolysis
32503
Alcaligenes faecalis
4-fluorophenylacetic acid + NH3
-
32503
Alcaligenes faecalis
-
4-fluorobenzylcyanide + H2O
i.e. p-fluorophenylacetonitrile, at 96% the rate of 4-chlorobenzylcyanide hydrolysis
32503
Alcaligenes faecalis JM3
4-fluorophenylacetic acid + NH3
-
32503
Alcaligenes faecalis JM3
-
4-hydroxybenzylcyanide + H2O
at 29% the rate of 4-chlorobenzylcyanide hydrolysis
32503
Alcaligenes faecalis
4-hydroxyphenylacetic acid + NH3
-
-
-
-
4-methoxybenzylcyanide + H2O
at 62% the rate of 4-chlorobenzylcyanide hydrolysis
32503
Alcaligenes faecalis
4-methoxyphenylacetic acid + NH3
-
-
-
-
4-nitrobenzylcyanide + H2O
at 45% the rate of 4-chlorobenzylcyanide hydrolysis
32503
Alcaligenes faecalis
4-nitrophenylacetate + NH3
-
-
-
-
acrylonitrile + H2O
at 7% the rate of 4-chlorobenzylcyanide hydrolysis
32503
Alcaligenes faecalis
propenoic acid + NH3
-
-
-
-
Arylacetonitrile + H2O
pathway in nitrile catabolism, inducible enzyme
32503
Alcaligenes faecalis
?
-
-
-
-
benzylcyanide + H2O
at 53% the rate of 4-chlorobenzylcyanide hydrolysis
32503
Alcaligenes faecalis
phenylacetic acid + NH3
-
32503
Alcaligenes faecalis
-
m-tolylacetonitrile + H2O
at 32% the rate of 4-chlorobenzylcyanide hydrolysis
32503
Alcaligenes faecalis
m-tolylacetic acid + NH3
-
-
-
-
m-xylenedicyanide + H2O
at 2.7% the rate of 4-chlorobenzylcyanide hydrolysis
32503
Alcaligenes faecalis
?
-
-
-
-
mandelonitrile + 2 H2O
at 8% the rate of 4-chlorobenzylcyanide hydrolysis
32503
Alcaligenes faecalis
2-hydroxyphenylacetic acid + NH3
-
-
-
-
additional information
very poor substrates (less than 1.5% the rate of 4-chlorobenzylcyanide hydrolysis) are 1,4-dichlorobenzylcyanide, 4-nitrobenzylcyanide, 2-pyridineacetonitrile, 2-phenylpropionitrile, 1-naphthylacetonitrile, diphenylacetonitrile, o-xylenedicyanide, acetonitrile (i.e. methylcyanide). No substrates are 2,6-dichlorobenzylcyanide, allylcyanide, propionitrile, aminoacetonitrile, iminodiacetonitrile, methacrylonitrile, crotonitrile, benzonitrile, 2- or 3-cyanopyridine, cyanopyrazine, 2-thiophenecarbonitrile or 2-furaneacrylonitrile
32503
Alcaligenes faecalis
?
-
-
-
-
additional information
very poor substrates (less than 1.5% the rate of 4-chlorobenzylcyanide hydrolysis) are 1,4-dichlorobenzylcyanide, 4-nitrobenzylcyanide, 2-pyridineacetonitrile, 2-phenylpropionitrile, 1-naphthylacetonitrile, diphenylacetonitrile, o-xylenedicyanide, acetonitrile (i.e. methylcyanide). No substrates are 2,6-dichlorobenzylcyanide, allylcyanide, propionitrile, aminoacetonitrile, iminodiacetonitrile, methacrylonitrile, crotonitrile, benzonitrile, 2- or 3-cyanopyridine, cyanopyrazine, 2-thiophenecarbonitrile or 2-furaneacrylonitrile
32503
Alcaligenes faecalis JM3
?
-
-
-
-
o-tolylacetonitrile + H2O
at 4% the rate of 4-chlorobenzylcyanide hydrolysis
32503
Alcaligenes faecalis
o-tolylacetic acid + NH3
-
-
-
-
p-tolylacetonitrile + H2O
at 72% the rate of 4-chlorobenzylcyanide hydrolysis
32503
Alcaligenes faecalis
p-tolylacetic acid + NH3
-
32503
Alcaligenes faecalis
-
Subunits
Subunits
Commentary
Organism
hexamer
6 * 44000, Alcaligenes faecalis, SDS-PAGE, 6 * 47770, Alcaligenes faecalis, minimal value derived from amino acid composition
Alcaligenes faecalis
Temperature Optimum [°C]
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
25
30
assay at
Alcaligenes faecalis
45
-
-
Alcaligenes faecalis
Temperature Range [°C]
Temperature Minimum [°C]
Temperature Maximum [°C]
Commentary
Organism
additional information
-
at 50°C and above, rapid loss of activity
Alcaligenes faecalis
Temperature Stability [°C]
Temperature Stability Minimum [°C]
Temperature Stability Maximum [°C]
Commentary
Organism
30
-
and below, 30 min, stable in 10 mM potassium phosphate buffer, pH 7.5
Alcaligenes faecalis
35
-
30 min, in 10 mM potassium phosphate buffer, pH 7.5, 25% loss of activity
Alcaligenes faecalis
40
-
30 min, in 10 mM potassium phosphate buffer, pH 7.5, inactivation
Alcaligenes faecalis
pH Optimum
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
7.5
-
2-thiopheneacetonitrile as substrate
Alcaligenes faecalis
pH Range
pH Minimum
pH Maximum
Commentary
Organism
6.4
9.4
about half-maximal activity at pH 6.4 and 9.4, 2-thiopheneacetonitrile as substrate
Alcaligenes faecalis
Activating Compound (protein specific)
Activating Compound
Commentary
Organism
Structure
2-mercaptoethanol
activation
Alcaligenes faecalis
dithiothreitol
activation
Alcaligenes faecalis
Sodium hydrosulfide
activation
Alcaligenes faecalis
Inhibitors (protein specific)
Inhibitors
Commentary
Organism
Structure
2,2'-dipyridyl
weak
Alcaligenes faecalis
3-Methyl-2-benzothiazolinone hydrazone
weak
Alcaligenes faecalis
8-hydroxyquinoline
weak
Alcaligenes faecalis
Ag2+
strong
Alcaligenes faecalis
ascorbic acid
-
Alcaligenes faecalis
Cd2+
-
Alcaligenes faecalis
Co2+
-
Alcaligenes faecalis
Cu2+
strong
Alcaligenes faecalis
D-cycloserine
weak
Alcaligenes faecalis
diethyldithiocarbamate
-
Alcaligenes faecalis
Disodium 4,5-dihydroxy-m-benzene-disulfonate
weak
Alcaligenes faecalis
DTNB
strong
Alcaligenes faecalis
H2O2
-
Alcaligenes faecalis
Hg2+
strong
Alcaligenes faecalis
hydroxylamine
weak
Alcaligenes faecalis
iodoacetate
weak
Alcaligenes faecalis
KCN
weak
Alcaligenes faecalis
L-cysteine
-
Alcaligenes faecalis
additional information
little or no inhibition by Li+, Na+, Mg2+, Ca2+, Ba2+, Mn2+, Fe2+, Pb2+, Fe3+, Al3+, NaN3, NaF, cysteamine, aminoguanidine, L/D-penicillamine, 1,10-phenanthroline or EDTA
Alcaligenes faecalis
N-ethylmaleimide
-
Alcaligenes faecalis
Ni2+
-
Alcaligenes faecalis
p-chloromercuribenzoate
strong
Alcaligenes faecalis
phenylhydrazine
-
Alcaligenes faecalis
Semicarbazide
weak
Alcaligenes faecalis
Sn2+
-
Alcaligenes faecalis
Urea
weak
Alcaligenes faecalis
Zn2+
-
Alcaligenes faecalis
KM Value [mM] (protein specific)
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.009
-
3-Indoleacetonitrile
-
Alcaligenes faecalis
0.01
-
benzylcyanide
-
Alcaligenes faecalis
0.015
-
2-Chlorobenzylcyanide
-
Alcaligenes faecalis
0.016
-
3-Thiopheneacetonitrile
-
Alcaligenes faecalis
0.022
-
4-Aminobenzylcyanide
-
Alcaligenes faecalis
0.023
-
3-tolylacetonitrile
-
Alcaligenes faecalis
0.065
-
3-Chlorobenzylcyanide
-
Alcaligenes faecalis
0.091
-
2-Thiopheneacetonitrile
-
Alcaligenes faecalis
0.105
-
4-Nitrobenzylcyanide
-
Alcaligenes faecalis
0.123
-
4-tolylacetonitrile
-
Alcaligenes faecalis
0.13
-
4-Fluorobenzylcyanide
-
Alcaligenes faecalis
0.333
-
4-Chlorobenzylcyanide
-
Alcaligenes faecalis
0.66
-
acrylonitrile
-
Alcaligenes faecalis
Molecular Weight [Da] (protein specific)
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
additional information
-
amino acid composition
Alcaligenes faecalis
44000
-
6 * 44000, Alcaligenes faecalis, SDS-PAGE, 6 * 47770, Alcaligenes faecalis, minimal value derived from amino acid composition
Alcaligenes faecalis
47770
-
6 * 44000, Alcaligenes faecalis, SDS-PAGE, 6 * 47770, Alcaligenes faecalis, minimal value derived from amino acid composition
Alcaligenes faecalis
260000
-
Alcaligenes faecalis, gel filtration on Sepharose 6B
Alcaligenes faecalis
275000
-
Alcaligenes faecalis, gel permeation HPLC
Alcaligenes faecalis
Natural Substrates/ Products (Substrates) (protein specific)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
Arylacetonitrile + H2O
Alcaligenes faecalis
pathway in nitrile catabolism, inducible enzyme
?
-
-
-
Organic Solvent Stability (protein specific)
Organic Solvent
Commentary
Organism
Methanol
stable in the presence of 10% w/v methanol
Alcaligenes faecalis
Purification (Commentary) (protein specific)
Commentary
Organism
-
Alcaligenes faecalis
Specific Activity [micromol/min/mg] (protein specific)
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
144
-
-
Alcaligenes faecalis
Storage Stability (protein specific)
Storage Stability
Organism
-20°C, in 10 mM potassium phosphate buffer, pH 7, 10 mM DTT and 50% glycerol, at least 5 months
Alcaligenes faecalis
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
Alcaligenes faecalis
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
Alcaligenes faecalis
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
Alcaligenes faecalis
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
1-cyclohexenylacetonitrile + H2O
at 3.3% the rate of 4-chlorobenzylcyanide hydrolysis
32503
Alcaligenes faecalis
1-cyclohexenylacetate + NH3
-
-
-
-
2,4-dichlorobenzylcyanide + H2O
at 6.5% the rate of 4-chlorobenzylcyanide hydrolysis
32503
Alcaligenes faecalis
2,4-dichlorophenylacetic acid + NH3
-
-
-
-
2-chlorobenzylcyanide + H2O
at 6% the rate of 4-chlorobenzylcyanide hydrolysis
32503
Alcaligenes faecalis
2-chlorophenylacetic acid + NH3
-
-
-
-
2-methoxybenzylcyanide + H2O
at 2% the rate of 4-chlorobenzylcyanide hydrolysis
32503
Alcaligenes faecalis
2-methoxyphenylacetic acid + NH3
-
-
-
-
2-thiopheneacetonitrile + H2O
ir, at 90% the rate of 4-chlorobenzylcyanide hydrolysis
32503
Alcaligenes faecalis
2-thiopheneacetic acid + NH3
not via thiopheneacetamide
32503
Alcaligenes faecalis
-
3,4-(methylenedioxy)phenylacetonitrile + H2O
at 65% the rate of 4-chlorobenzylcyanide hydrolysis
32503
Alcaligenes faecalis
3,4-(methylendioxy)phenylacetic acid + NH3
-
-
-
-
3,4-dichlorobenzylcyanide + H2O
at 14% the rate of 4-chlorobenzylcyanide hydrolysis
32503
Alcaligenes faecalis
3,4-dichlorophenylacetic acid + NH3
-
-
-
-
3-chlorobenzylcyanide + H2O
at 43% the rate of 4-chlorobenzylcyanide hydrolysis
32503
Alcaligenes faecalis
3-chlorophenylacetic acid + NH3
-
-
-
-
3-indoleacetonitrile + H2O
at 9% the rate of 4-chlorobenzylcyanide hydrolysis
32503
Alcaligenes faecalis
3-indoleacetic acid + NH3
-
32503
Alcaligenes faecalis
-
3-indoleacetonitrile + H2O
at 9% the rate of 4-chlorobenzylcyanide hydrolysis
32503
Alcaligenes faecalis JM3
3-indoleacetic acid + NH3
-
32503
Alcaligenes faecalis JM3
-
3-phenylpropionitrile + H2O
at 26% the rate of 4-chlorobenzylcyanide hydrolysis
32503
Alcaligenes faecalis
3-phenylpropionic acid + NH3
-
-
-
-
3-pyridineacetonitrile + H2O
at 77% the rate of 4-chlorobenzylcyanide hydrolysis
32503
Alcaligenes faecalis
?
-
-
-
-
3-thiopheneacetonitrile + H2O
at 24% the rate of 4-chlorobenzylcyanide hydrolysis
32503
Alcaligenes faecalis
3-thiopheneacetic acid + NH3
-
32503
Alcaligenes faecalis
-
4-aminobenzylcyanide + H2O
at 44% the rate of 4-chlorobenzylcyanide hydrolysis
32503
Alcaligenes faecalis
4-aminophenylacetic acid + NH3
-
-
-
-
4-bromobenzylcyanide + H2O
at 26% the rate of 4-chlorobenzylcyanide hydrolysis
32503
Alcaligenes faecalis
?
-
-
-
-
4-bromobenzylcyanide + H2O
at 26% the rate of 4-chlorobenzylcyanide hydrolysis
32503
Alcaligenes faecalis JM3
?
-
-
-
-
4-chlorobenzylcyanide + H2O
i.e. p-chlorophenylacetonitrile, best substrate
32503
Alcaligenes faecalis
4-chlorophenylacetic acid + NH3
-
32503
Alcaligenes faecalis
-
4-chlorobenzylcyanide + H2O
i.e. p-chlorophenylacetonitrile, best substrate
32503
Alcaligenes faecalis JM3
4-chlorophenylacetic acid + NH3
-
32503
Alcaligenes faecalis JM3
-
4-fluorobenzylcyanide + H2O
i.e. p-fluorophenylacetonitrile, at 96% the rate of 4-chlorobenzylcyanide hydrolysis
32503
Alcaligenes faecalis
4-fluorophenylacetic acid + NH3
-
32503
Alcaligenes faecalis
-
4-fluorobenzylcyanide + H2O
i.e. p-fluorophenylacetonitrile, at 96% the rate of 4-chlorobenzylcyanide hydrolysis
32503
Alcaligenes faecalis JM3
4-fluorophenylacetic acid + NH3
-
32503
Alcaligenes faecalis JM3
-
4-hydroxybenzylcyanide + H2O
at 29% the rate of 4-chlorobenzylcyanide hydrolysis
32503
Alcaligenes faecalis
4-hydroxyphenylacetic acid + NH3
-
-
-
-
4-methoxybenzylcyanide + H2O
at 62% the rate of 4-chlorobenzylcyanide hydrolysis
32503
Alcaligenes faecalis
4-methoxyphenylacetic acid + NH3
-
-
-
-
4-nitrobenzylcyanide + H2O
at 45% the rate of 4-chlorobenzylcyanide hydrolysis
32503
Alcaligenes faecalis
4-nitrophenylacetate + NH3
-
-
-
-
acrylonitrile + H2O
at 7% the rate of 4-chlorobenzylcyanide hydrolysis
32503
Alcaligenes faecalis
propenoic acid + NH3
-
-
-
-
Arylacetonitrile + H2O
pathway in nitrile catabolism, inducible enzyme
32503
Alcaligenes faecalis
?
-
-
-
-
benzylcyanide + H2O
at 53% the rate of 4-chlorobenzylcyanide hydrolysis
32503
Alcaligenes faecalis
phenylacetic acid + NH3
-
32503
Alcaligenes faecalis
-
m-tolylacetonitrile + H2O
at 32% the rate of 4-chlorobenzylcyanide hydrolysis
32503
Alcaligenes faecalis
m-tolylacetic acid + NH3
-
-
-
-
m-xylenedicyanide + H2O
at 2.7% the rate of 4-chlorobenzylcyanide hydrolysis
32503
Alcaligenes faecalis
?
-
-
-
-
mandelonitrile + 2 H2O
at 8% the rate of 4-chlorobenzylcyanide hydrolysis
32503
Alcaligenes faecalis
2-hydroxyphenylacetic acid + NH3
-
-
-
-
additional information
very poor substrates (less than 1.5% the rate of 4-chlorobenzylcyanide hydrolysis) are 1,4-dichlorobenzylcyanide, 4-nitrobenzylcyanide, 2-pyridineacetonitrile, 2-phenylpropionitrile, 1-naphthylacetonitrile, diphenylacetonitrile, o-xylenedicyanide, acetonitrile (i.e. methylcyanide). No substrates are 2,6-dichlorobenzylcyanide, allylcyanide, propionitrile, aminoacetonitrile, iminodiacetonitrile, methacrylonitrile, crotonitrile, benzonitrile, 2- or 3-cyanopyridine, cyanopyrazine, 2-thiophenecarbonitrile or 2-furaneacrylonitrile
32503
Alcaligenes faecalis
?
-
-
-
-
additional information
very poor substrates (less than 1.5% the rate of 4-chlorobenzylcyanide hydrolysis) are 1,4-dichlorobenzylcyanide, 4-nitrobenzylcyanide, 2-pyridineacetonitrile, 2-phenylpropionitrile, 1-naphthylacetonitrile, diphenylacetonitrile, o-xylenedicyanide, acetonitrile (i.e. methylcyanide). No substrates are 2,6-dichlorobenzylcyanide, allylcyanide, propionitrile, aminoacetonitrile, iminodiacetonitrile, methacrylonitrile, crotonitrile, benzonitrile, 2- or 3-cyanopyridine, cyanopyrazine, 2-thiophenecarbonitrile or 2-furaneacrylonitrile
32503
Alcaligenes faecalis JM3
?
-
-
-
-
o-tolylacetonitrile + H2O
at 4% the rate of 4-chlorobenzylcyanide hydrolysis
32503
Alcaligenes faecalis
o-tolylacetic acid + NH3
-
-
-
-
p-tolylacetonitrile + H2O
at 72% the rate of 4-chlorobenzylcyanide hydrolysis
32503
Alcaligenes faecalis
p-tolylacetic acid + NH3
-
32503
Alcaligenes faecalis
-
Subunits (protein specific)
Subunits
Commentary
Organism
hexamer
6 * 44000, Alcaligenes faecalis, SDS-PAGE, 6 * 47770, Alcaligenes faecalis, minimal value derived from amino acid composition
Alcaligenes faecalis
Temperature Optimum [°C] (protein specific)
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
25
30
assay at
Alcaligenes faecalis
45
-
-
Alcaligenes faecalis
Temperature Range [°C] (protein specific)
Temperature Minimum [°C]
Temperature Maximum [°C]
Commentary
Organism
additional information
-
at 50°C and above, rapid loss of activity
Alcaligenes faecalis
Temperature Stability [°C] (protein specific)
Temperature Stability Minimum [°C]
Temperature Stability Maximum [°C]
Commentary
Organism
30
-
and below, 30 min, stable in 10 mM potassium phosphate buffer, pH 7.5
Alcaligenes faecalis
35
-
30 min, in 10 mM potassium phosphate buffer, pH 7.5, 25% loss of activity
Alcaligenes faecalis
40
-
30 min, in 10 mM potassium phosphate buffer, pH 7.5, inactivation
Alcaligenes faecalis
pH Optimum (protein specific)
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
7.5
-
2-thiopheneacetonitrile as substrate
Alcaligenes faecalis
pH Range (protein specific)
pH Minimum
pH Maximum
Commentary
Organism
6.4
9.4
about half-maximal activity at pH 6.4 and 9.4, 2-thiopheneacetonitrile as substrate
Alcaligenes faecalis
Other publictions for EC 3.5.5.5
No.
1st author
Pub Med
title
organims
journal
volume
pages
year
Activating Compound
Application
Cloned(Commentary)
Crystallization (Commentary)
Engineering
General Stability
Inhibitors
KM Value [mM]
Localization
Metals/Ions
Molecular Weight [Da]
Natural Substrates/ Products (Substrates)
Organic Solvent Stability
Organism
Oxidation Stability
Posttranslational Modification
Purification (Commentary)
Reaction
Renatured (Commentary)
Source Tissue
Specific Activity [micromol/min/mg]
Storage Stability
Substrates and Products (Substrate)
Subunits
Temperature Optimum [°C]
Temperature Range [°C]
Temperature Stability [°C]
Turnover Number [1/s]
pH Optimum
pH Range
pH Stability
Cofactor
Ki Value [mM]
pI Value
IC50 Value
Activating Compound (protein specific)
Application (protein specific)
Cloned(Commentary) (protein specific)
Cofactor (protein specific)
Crystallization (Commentary) (protein specific)
Engineering (protein specific)
General Stability (protein specific)
IC50 Value (protein specific)
Inhibitors (protein specific)
Ki Value [mM] (protein specific)
KM Value [mM] (protein specific)
Localization (protein specific)
Metals/Ions (protein specific)
Molecular Weight [Da] (protein specific)
Natural Substrates/ Products (Substrates) (protein specific)
Organic Solvent Stability (protein specific)
Oxidation Stability (protein specific)
Posttranslational Modification (protein specific)
Purification (Commentary) (protein specific)
Renatured (Commentary) (protein specific)
Source Tissue (protein specific)
Specific Activity [micromol/min/mg] (protein specific)
Storage Stability (protein specific)
Substrates and Products (Substrate) (protein specific)
Subunits (protein specific)
Temperature Optimum [°C] (protein specific)
Temperature Range [°C] (protein specific)
Temperature Stability [°C] (protein specific)
Turnover Number [1/s] (protein specific)
pH Optimum (protein specific)
pH Range (protein specific)
pH Stability (protein specific)
pI Value (protein specific)
Expression
General Information
General Information (protein specific)
Expression (protein specific)
KCat/KM [mM/s]
KCat/KM [mM/s] (protein specific)
733193
Sosedov
Improvement of the amides form ...
Pseudomonas fluorescens, Pseudomonas fluorescens EBC191
Appl. Microbiol. Biotechnol.
99
2623-2635
2015
-
-
1
-
1
-
-
-
-
-
-
-
-
11
-
-
-
-
-
-
-
-
4
-
-
-
-
-
-
-
-
-
-
-
-
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1
-
-
1
-
-
-
-
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-
-
-
-
-
-
-
-
-
-
-
-
4
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
733558
Sun
Cloning, purification and eval ...
Luminiphilus syltensis, Luminiphilus syltensis NOR5-1B
Biotechnol. Lett.
37
1655-1661
2015
-
-
1
-
-
-
-
1
-
-
1
-
-
4
-
-
1
-
-
-
-
-
41
1
1
1
1
-
1
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
1
-
-
1
-
-
-
-
1
-
-
-
-
41
1
1
1
1
-
1
-
-
-
-
-
-
-
-
-
733546
Bhatia
Purification and characterizat ...
Alcaligenes sp., Alcaligenes sp. MTCC 10675
Biotechnol. Appl. Biochem.
61
459-465
2014
-
-
-
-
-
1
10
1
-
1
2
-
-
3
-
-
1
-
-
-
-
-
8
1
1
-
1
1
1
-
-
-
-
-
-
-
-
-
-
-
-
1
-
10
-
1
-
1
2
-
-
-
-
1
-
-
-
-
8
1
1
-
1
1
1
-
-
-
-
-
-
-
-
-
718648
Baum
Conversion of sterically deman ...
Pseudomonas fluorescens, Pseudomonas fluorescens EBC191
Appl. Environ. Microbiol.
78
48-57
2012
-
1
1
-
11
-
-
1
-
-
-
2
-
10
-
-
-
-
-
-
-
-
14
-
1
-
-
-
2
-
-
-
-
-
-
-
1
1
-
-
11
-
-
-
-
1
-
-
-
2
-
-
-
-
-
-
-
-
14
-
1
-
-
-
2
-
-
-
-
-
-
-
-
-
718670
Petrickova
Purification and characterizat ...
Aspergillus niger, Aspergillus niger CBS 513.88, Neurospora crassa, Neurospora crassa OR74A
Appl. Microbiol. Biotechnol.
93
1553-1561
2012
-
-
2
-
-
-
-
6
-
-
-
6
-
7
-
-
2
-
-
-
-
-
24
-
2
-
8
-
2
-
9
-
-
-
-
-
-
3
-
-
-
-
-
-
-
9
-
-
-
6
-
-
-
3
-
-
-
-
24
-
3
-
12
-
3
-
14
-
-
1
1
-
-
-
734496
Petrickova
-
Influence of point mutations n ...
Aspergillus niger, Aspergillus niger CBS 513.88, Neurospora crassa, Neurospora crassa DSM 1257
J. Mol. Catal. B
77
74-80
2012
-
-
2
-
4
-
-
-
-
-
-
-
-
5
-
-
-
-
-
-
-
-
8
-
-
-
-
-
2
4
-
-
-
-
-
-
-
2
-
-
4
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
8
-
-
-
-
-
2
4
-
-
-
-
-
-
-
-
719297
Detzel
-
Autodisplay of nitrilase from ...
Alcaligenes faecalis, Alcaligenes faecalis ATCC 8750
ChemCatChem
3
719-725
2011
-
-
1
-
-
-
-
-
-
-
-
-
-
4
-
-
-
-
-
-
-
-
2
-
1
-
-
-
1
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
2
-
1
-
-
-
1
-
-
-
-
-
-
-
-
-
719629
Liu
Gene cloning, expression, and ...
Alcaligenes faecalis, Alcaligenes faecalis ZJUTB10
J. Agric. Food Chem.
59
11560-11570
2011
-
1
1
-
3
-
11
1
-
2
1
2
-
2
-
-
1
1
-
-
-
-
12
2
1
1
1
-
1
-
-
-
-
-
-
-
1
1
-
-
3
-
-
11
-
1
-
2
1
2
-
-
-
1
-
-
-
-
12
2
1
1
1
-
1
-
-
-
-
1
1
-
-
-
710940
Sosedov
Construction and application o ...
Pseudomonas fluorescens
Appl. Environ. Microbiol.
76
3668-3674
2010
-
-
-
-
14
-
-
-
-
-
-
-
-
5
-
-
-
-
-
-
-
-
2
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
14
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
2
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
712643
He
Biocatalytic synthesis of (R)- ...
Alcaligenes faecalis
J. Ind. Microbiol. Biotechnol.
37
741-750
2010
-
-
-
-
-
-
-
-
-
-
-
4
-
2
-
-
-
-
-
-
-
-
5
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
4
-
-
-
-
-
-
-
-
5
-
-
-
-
-
-
-
-
-
1
-
-
1
-
-
710930
Kiziak
Identification of amino acid r ...
Pseudomonas fluorescens
Appl. Environ. Microbiol.
75
5592-5599
2009
-
-
1
-
8
-
-
-
-
-
-
-
-
5
-
-
-
-
-
-
-
-
2
-
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
8
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
2
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
712260
Thuku
Microbial nitrilases: Versatil ...
Alcaligenes faecalis, Alcaligenes faecalis ATCC 8750
J. Appl. Microbiol.
106
703-727
2009
-
-
-
-
-
-
-
-
-
-
2
-
-
4
-
-
-
-
-
-
-
-
6
1
1
-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
2
-
-
-
-
-
-
-
-
-
6
1
1
-
-
-
1
-
-
-
-
-
-
-
-
-
713073
Howden
Pseudomonas syringae pv. syrin ...
no activity in no activity in Pseudomonas syringae pv. tomato, Pseudomonas syringae pv. syringae, Pseudomonas syringae pv. syringae B728a
Mol. Plant Pathol.
10
857-865
2009
-
-
-
-
-
-
-
-
-
-
-
-
-
10
-
-
-
-
-
-
-
-
4
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-
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-
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-
-
-
4
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
717051
Sosedov
-
Synthesis of enantiomerically ...
Pseudomonas fluorescens, Pseudomonas fluorescens EBC191
Adv. Synth. Catal.
351
1531 - 1538
2009
-
-
1
-
-
-
-
-
-
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5
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2
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1
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-
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-
-
2
-
-
-
-
-
-
-
-
-
-
1
1
-
-
-
684655
Rustler
Simultaneous expression of an ...
Pseudomonas fluorescens, Pseudomonas fluorescens EBC191
Appl. Microbiol. Biotechnol.
80
87-97
2008
-
-
1
-
-
-
-
-
-
-
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-
-
13
-
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-
-
-
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2
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1
1
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1
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-
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-
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-
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-
-
-
2
-
-
-
-
-
-
1
1
-
-
-
-
-
-
-
687882
Zhu
A new nitrilase from Bradyrhiz ...
Bradyrhizobium japonicum, Bradyrhizobium japonicum USDA 110
J. Biotechnol.
133
327-333
2008
-
-
1
-
-
-
-
1
-
-
3
-
-
24
-
-
1
-
-
-
-
-
26
1
1
-
1
1
1
1
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
1
-
-
3
-
-
-
-
1
-
-
-
-
26
1
1
-
1
1
1
1
-
-
-
-
-
-
-
-
694107
Banerjee
enantioselective nitrilase fro ...
Pseudomonas putida, Pseudomonas putida MTCC 5110
Mol. Biotechnol.
41
35-41
2008
-
1
1
-
-
-
-
-
-
-
1
2
-
2
-
-
1
-
-
-
10
-
2
-
1
-
-
-
1
-
-
-
-
-
-
-
1
1
-
-
-
-
-
-
-
-
-
-
1
2
-
-
-
1
-
-
10
-
2
-
1
-
-
-
1
-
-
-
-
-
-
-
-
-
700670
Agerbirk
Sinapis phylogeny and evolutio ...
Sinapis alba
Phytochemistry
69
2937-2949
2008
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
-
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-
-
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3
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-
-
-
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-
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-
-
-
-
-
3
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
701254
Rustler
Characterisation of the substr ...
Exophiala oligosperma, Exophiala oligosperma R1
Stud. Mycol.
61
165-174
2008
1
-
-
-
-
-
-
-
-
-
-
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2
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-
-
-
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12
-
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-
-
-
-
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1
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-
-
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-
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-
-
-
-
-
-
12
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
689852
Kiziak
Influence of different carboxy ...
Pseudomonas fluorescens, Pseudomonas fluorescens EBC191
Protein Eng. Des. Sel.
20
385-396
2007
-
-
1
-
1
-
-
-
-
-
-
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10
-
-
-
-
-
-
1
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4
-
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1
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1
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-
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-
-
1
-
4
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
667345
Banerjee
Purification and characterizat ...
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18
2
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3
4
1
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4
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7
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3
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3
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1
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1
2
18
2
1
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3
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1
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2
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2
15
1
1
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1
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1
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1
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1
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1
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2
15
1
1
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5
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1
4
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1
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20
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33
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1
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1
1
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3
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27
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13
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5
1
1
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1
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1
4
33
1
2
1
3
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1
1
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