Literature summary for 3.5.5.5 extracted from

  • Nagasawa, T.; Mauger, J.; Yamada, H.
    A novel nitrilase, arylacetonitrilase, of Alcaligenes faecalis JM3. Purification and characterization (1990), Eur. J. Biochem., 194, 765-772.
    View publication on PubMed

Activating Compound

Activating Compound Comment Organism Structure
dithiothreitol activation Alcaligenes faecalis
2-mercaptoethanol activation Alcaligenes faecalis
Sodium hydrosulfide activation Alcaligenes faecalis

Inhibitors

Inhibitors Comment 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 Comment Organism Structure
0.015
-
2-Chlorobenzylcyanide
-
Alcaligenes faecalis
0.022
-
4-Aminobenzylcyanide
-
Alcaligenes faecalis
0.065
-
3-Chlorobenzylcyanide
-
Alcaligenes faecalis
0.009
-
3-Indoleacetonitrile
-
Alcaligenes faecalis
0.023
-
3-tolylacetonitrile
-
Alcaligenes faecalis
0.01
-
benzylcyanide
-
Alcaligenes faecalis
0.13
-
4-Fluorobenzylcyanide
-
Alcaligenes faecalis
0.66
-
acrylonitrile
-
Alcaligenes faecalis
0.091
-
2-Thiopheneacetonitrile
-
Alcaligenes faecalis
0.016
-
3-Thiopheneacetonitrile
-
Alcaligenes faecalis
0.105
-
4-Nitrobenzylcyanide
-
Alcaligenes faecalis
0.333
-
4-Chlorobenzylcyanide
-
Alcaligenes faecalis
0.123
-
4-tolylacetonitrile
-
Alcaligenes faecalis

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
additional information
-
amino acid composition Alcaligenes faecalis
260000
-
Alcaligenes faecalis, gel filtration on Sepharose 6B Alcaligenes faecalis
44000
-
6 * 44000, Alcaligenes faecalis, SDS-PAGE, 6 * 47770, Alcaligenes faecalis, minimal value derived from amino acid composition Alcaligenes faecalis
275000
-
Alcaligenes faecalis, gel permeation HPLC Alcaligenes faecalis
47770
-
6 * 44000, Alcaligenes faecalis, SDS-PAGE, 6 * 47770, Alcaligenes faecalis, minimal value derived from amino acid composition Alcaligenes faecalis

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
Arylacetonitrile + H2O Alcaligenes faecalis pathway in nitrile catabolism, inducible enzyme ?
-
?

Organic Solvent Stability

Organic Solvent Comment Organism
Methanol stable in the presence of 10% w/v methanol Alcaligenes faecalis

Organism

Organism UniProt Comment Textmining
Alcaligenes faecalis
-
JM3
-
Alcaligenes faecalis JM3
-
JM3
-

Purification (Commentary)

Purification (Comment) Organism
-
Alcaligenes faecalis

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment 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 Comment Substrates Organism Products Comment (Products) Rev. Reac.
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 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 Alcaligenes faecalis JM3 ?
-
?
4-chlorobenzylcyanide + H2O i.e. p-chlorophenylacetonitrile, best substrate Alcaligenes faecalis 4-chlorophenylacetic acid + NH3
-
?
4-chlorobenzylcyanide + H2O i.e. p-chlorophenylacetonitrile, best substrate Alcaligenes faecalis JM3 4-chlorophenylacetic acid + NH3
-
?
4-fluorobenzylcyanide + H2O i.e. p-fluorophenylacetonitrile, at 96% the rate of 4-chlorobenzylcyanide hydrolysis Alcaligenes faecalis 4-fluorophenylacetic acid + NH3
-
?
4-fluorobenzylcyanide + H2O i.e. p-fluorophenylacetonitrile, at 96% the rate of 4-chlorobenzylcyanide hydrolysis Alcaligenes faecalis JM3 4-fluorophenylacetic acid + NH3
-
?
4-bromobenzylcyanide + H2O at 26% the rate of 4-chlorobenzylcyanide hydrolysis Alcaligenes faecalis ?
-
?
4-bromobenzylcyanide + H2O at 26% the rate of 4-chlorobenzylcyanide hydrolysis Alcaligenes faecalis JM3 ?
-
?
3-indoleacetonitrile + H2O at 9% the rate of 4-chlorobenzylcyanide hydrolysis Alcaligenes faecalis 3-indoleacetic acid + NH3
-
?
3-indoleacetonitrile + H2O at 9% the rate of 4-chlorobenzylcyanide hydrolysis Alcaligenes faecalis JM3 3-indoleacetic acid + NH3
-
?
2-thiopheneacetonitrile + H2O ir, at 90% the rate of 4-chlorobenzylcyanide hydrolysis Alcaligenes faecalis 2-thiopheneacetic acid + NH3 not via thiopheneacetamide ?
3-thiopheneacetonitrile + H2O at 24% the rate of 4-chlorobenzylcyanide hydrolysis Alcaligenes faecalis 3-thiopheneacetic acid + NH3
-
?
benzylcyanide + H2O at 53% the rate of 4-chlorobenzylcyanide hydrolysis Alcaligenes faecalis phenylacetic acid + NH3
-
?
p-tolylacetonitrile + H2O at 72% the rate of 4-chlorobenzylcyanide hydrolysis Alcaligenes faecalis p-tolylacetic acid + NH3
-
?
m-tolylacetonitrile + H2O at 32% the rate of 4-chlorobenzylcyanide hydrolysis Alcaligenes faecalis m-tolylacetic acid + NH3
-
?
o-tolylacetonitrile + H2O at 4% the rate of 4-chlorobenzylcyanide hydrolysis Alcaligenes faecalis o-tolylacetic acid + NH3
-
?
3-chlorobenzylcyanide + H2O at 43% the rate of 4-chlorobenzylcyanide hydrolysis Alcaligenes faecalis 3-chlorophenylacetic acid + NH3
-
?
2-chlorobenzylcyanide + H2O at 6% the rate of 4-chlorobenzylcyanide hydrolysis Alcaligenes faecalis 2-chlorophenylacetic acid + NH3
-
?
4-aminobenzylcyanide + H2O at 44% the rate of 4-chlorobenzylcyanide hydrolysis Alcaligenes faecalis 4-aminophenylacetic acid + NH3
-
?
4-nitrobenzylcyanide + H2O at 45% the rate of 4-chlorobenzylcyanide hydrolysis Alcaligenes faecalis 4-nitrophenylacetate + NH3
-
?
4-hydroxybenzylcyanide + H2O at 29% the rate of 4-chlorobenzylcyanide hydrolysis Alcaligenes faecalis 4-hydroxyphenylacetic acid + NH3
-
?
4-methoxybenzylcyanide + H2O at 62% the rate of 4-chlorobenzylcyanide hydrolysis Alcaligenes faecalis 4-methoxyphenylacetic acid + NH3
-
?
2-methoxybenzylcyanide + H2O at 2% the rate of 4-chlorobenzylcyanide hydrolysis Alcaligenes faecalis 2-methoxyphenylacetic acid + NH3
-
?
3,4-dichlorobenzylcyanide + H2O at 14% the rate of 4-chlorobenzylcyanide hydrolysis Alcaligenes faecalis 3,4-dichlorophenylacetic acid + NH3
-
?
2,4-dichlorobenzylcyanide + H2O at 6.5% the rate of 4-chlorobenzylcyanide hydrolysis Alcaligenes faecalis 2,4-dichlorophenylacetic acid + NH3
-
?
3-pyridineacetonitrile + H2O at 77% the rate of 4-chlorobenzylcyanide hydrolysis Alcaligenes faecalis ?
-
?
mandelonitrile + 2 H2O at 8% the rate of 4-chlorobenzylcyanide hydrolysis Alcaligenes faecalis mandelic acid + NH3
-
?
3,4-(methylenedioxy)phenylacetonitrile + H2O at 65% the rate of 4-chlorobenzylcyanide hydrolysis Alcaligenes faecalis 3,4-(methylendioxy)phenylacetic acid + NH3
-
?
3-phenylpropionitrile + H2O at 26% the rate of 4-chlorobenzylcyanide hydrolysis Alcaligenes faecalis 3-phenylpropionic acid + NH3
-
?
m-xylenedicyanide + H2O at 2.7% the rate of 4-chlorobenzylcyanide hydrolysis Alcaligenes faecalis ?
-
?
acrylonitrile + H2O at 7% the rate of 4-chlorobenzylcyanide hydrolysis Alcaligenes faecalis propenoic acid + NH3
-
?
1-cyclohexenylacetonitrile + H2O at 3.3% the rate of 4-chlorobenzylcyanide hydrolysis Alcaligenes faecalis 1-cyclohexenylacetate + NH3
-
?
Arylacetonitrile + H2O pathway in nitrile catabolism, inducible enzyme Alcaligenes faecalis ?
-
?

Subunits

Subunits Comment 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] Comment Organism
45
-
-
Alcaligenes faecalis
25 30 assay at Alcaligenes faecalis

Temperature Range [°C]

Temperature Minimum [°C] Temperature Maximum [°C] Comment 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] Comment Organism
40
-
30 min, in 10 mM potassium phosphate buffer, pH 7.5, inactivation Alcaligenes faecalis
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

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.5
-
2-thiopheneacetonitrile as substrate Alcaligenes faecalis

pH Range

pH Minimum pH Maximum Comment Organism
6.4 9.4 about half-maximal activity at pH 6.4 and 9.4, 2-thiopheneacetonitrile as substrate Alcaligenes faecalis