Any feedback?
Please rate this page
(literature.php)
(0/150)

BRENDA support

Literature summary for 3.5.5.7 extracted from

  • Yeom, S.J.; Kim, H.J.; Lee, J.K.; Kim, D.E.; Oh, D.K.
    A determinant residue of substrate specificity in nitrilase from Rhodococcus rhodochrous ATCC 33278 for aliphatic and aromatic nitriles (2008), Biochem. J., 415, 401-407.
    View publication on PubMedView publication on EuropePMC
Search for more literature for 3.5.5.7

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli strain ER2566 Rhodococcus rhodochrous

Crystallization (Commentary)

Crystallization (Comment) Organism
molecular modeling of enzyme and docking studies using aromatic and aliphatic nitrile substrates Rhodococcus rhodochrous

Protein Variants

Protein Variants Comment Organism
K9A activity similar to wild-type Rhodococcus rhodochrous
K9A increased specific activities Rhodococcus rhodochrous
L79A increased specific activities Rhodococcus rhodochrous
L79A slight decrease in activity towards aromatic nitriles Rhodococcus rhodochrous
additional information mutants containing charged amino acids such as aspartate, glutamate, arginine and asparagine at position 142 display no activity towards any nitrile, possibly owing to the disruption of hydrophobic interactions with substrates Rhodococcus rhodochrous
R129A inactive Rhodococcus rhodochrous
R129A loss of enzymatic activiy towards all nitriles Rhodococcus rhodochrous
Y142A mutant exhibits slightly higher kcat/Km values for aromatic nitriles and shows no activity toward aliphatic nitriles Rhodococcus rhodochrous
Y142A loss of activity towards aliphatic nitriles Rhodococcus rhodochrous
Y142A mutant specific for aromatic nitrile substrates Rhodococcus rhodochrous
Y142D complete loss of activity Rhodococcus rhodochrous
Y142E complete loss of activity Rhodococcus rhodochrous
Y142F substrate specificity similar to wild-type Rhodococcus rhodochrous
Y142F mutant shows slightly lower kcat/Km values compared to the wild type enzyme Rhodococcus rhodochrous
Y142G complete loss of activity Rhodococcus rhodochrous
Y142I very low activity Rhodococcus rhodochrous
Y142I low specific activities Rhodococcus rhodochrous
Y142L low specific activities Rhodococcus rhodochrous
Y142L mutant specific for aromatic nitrile substrates Rhodococcus rhodochrous
Y142N complete loss of activity Rhodococcus rhodochrous
Y142R complete loss of activity Rhodococcus rhodochrous
Y142S mutant exhibits slightly higher kcat/Km values for aromatic nitriles and shows no activity toward aliphatic nitriles Rhodococcus rhodochrous
Y142S mutant specific for aromatic nitrile substrates Rhodococcus rhodochrous
Y142V mutant specific for aromatic nitrile substrates Rhodococcus rhodochrous
Y175A increased specific activities Rhodococcus rhodochrous
Y175A activity below 6 micromol/min mg Rhodococcus rhodochrous

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.47
-
 m-tolunitrile mutant Y142V, pH 7.5, 25°C Rhodococcus rhodochrous
0.49
-
 Benzonitrile mutant Y142V, pH 7.5, 25°C Rhodococcus rhodochrous
0.61
-
 m-tolunitrile mutant Y142A, pH 7.5, 25°C Rhodococcus rhodochrous
0.688
-
 adiponitrile mutant enzyme Y142F Rhodococcus rhodochrous
0.69
-
 adiponitrile mutant Y142F, pH 7.5, 25°C Rhodococcus rhodochrous
0.82
-
 2-Cyanopyridine mutant Y142A, pH 7.5, 25°C Rhodococcus rhodochrous
0.85
-
 adiponitrile mutant K9A, pH 7.5, 25°C Rhodococcus rhodochrous
0.91
-
 Benzonitrile mutant L79A, pH 7.5, 25°C Rhodococcus rhodochrous
0.92
-
 adiponitrile mutant L79A, pH 7.5, 25°C Rhodococcus rhodochrous
0.93
-
 adiponitrile wild-type, pH 7.5, 25°C Rhodococcus rhodochrous
0.933
-
 adiponitrile wild type enzyme Rhodococcus rhodochrous
1.02
-
 Benzonitrile mutant K9A, pH 7.5, 25°C Rhodococcus rhodochrous
1.07
-
 Benzonitrile wild-type, pH 7.5, 25°C Rhodococcus rhodochrous
1.15
-
 glutaronitrile mutant L79A, pH 7.5, 25°C Rhodococcus rhodochrous
1.19
-
 glutaronitrile mutant K9A, pH 7.5, 25°C Rhodococcus rhodochrous
1.19
-
 m-tolunitrile mutant Y142F, pH 7.5, 25°C Rhodococcus rhodochrous
1.24
-
 glutaronitrile wild-type, pH 7.5, 25°C Rhodococcus rhodochrous
1.241
-
 glutaronitrile wild type enzyme Rhodococcus rhodochrous
1.25
-
 2-Cyanopyridine mutant K9A, pH 7.5, 25°C Rhodococcus rhodochrous
1.49
-
 m-tolunitrile mutant Y142S, pH 7.5, 25°C Rhodococcus rhodochrous
1.5
-
 2-Cyanopyridine mutant L79A, pH 7.5, 25°C Rhodococcus rhodochrous
1.53
-
 2-Cyanopyridine wild-type, pH 7.5, 25°C Rhodococcus rhodochrous
1.56
-
 glutaronitrile mutant enzyme Y142F Rhodococcus rhodochrous
1.56
-
 2-Cyanopyridine mutant Y142F, pH 7.5, 25°C Rhodococcus rhodochrous
1.56
-
 glutaronitrile mutant Y142F, pH 7.5, 25°C Rhodococcus rhodochrous
1.58
-
 Benzonitrile mutant Y142A, pH 7.5, 25°C Rhodococcus rhodochrous
1.62
-
 m-tolunitrile mutant K9A, pH 7.5, 25°C Rhodococcus rhodochrous
1.79
-
 m-tolunitrile mutant L79A, pH 7.5, 25°C Rhodococcus rhodochrous
1.82
-
 Benzonitrile mutant Y142S, pH 7.5, 25°C Rhodococcus rhodochrous
1.85
-
 m-tolunitrile wild-type, pH 7.5, 25°C Rhodococcus rhodochrous
3.45
-
 Benzonitrile mutant Y142F, pH 7.5, 25°C Rhodococcus rhodochrous
3.7 5 2-Cyanopyridine mutant Y142S, pH 7.5, 25°C Rhodococcus rhodochrous

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
40000
-
 SDS-PAGE Rhodococcus rhodochrous

Organic Solvent Stability

Organic Solvent Comment Organism
Methanol activity assay in presence of 10% v/v Rhodococcus rhodochrous

Organism

Organism UniProt Comment Textmining
Rhodococcus rhodochrous
-
 strain ATCC 33278
-
Rhodococcus rhodochrous Q03217 strain ATCC 33278
-

Purification (Commentary)

Purification (Comment) Organism
HisTrap HP 9 column chromatography Rhodococcus rhodochrous

Reaction

Reaction Comment Organism Reaction ID
R-CN + 2 H2O = R-COOH + NH3 hydrolysis of substrates probably involves the conjugated pi-electron system of the aromatic ring of substrate or Tyr-142 as an electron acceptor Rhodococcus rhodochrous
a

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
1.4
-
 mutant enzyme Y175A, using succinonitrile as a substrate Rhodococcus rhodochrous
1.7
-
 wild type enzyme, using sebacconitrile as a substrate Rhodococcus rhodochrous
1.8
-
 mutant enzyme K9A, using sebacconitrile as a substrate Rhodococcus rhodochrous
3.9
-
 mutant enzyme Y175A, using glutaronitrile as a substrate Rhodococcus rhodochrous
4.3
-
 wild type enzyme, using succinonitrile as a substrate Rhodococcus rhodochrous
4.8
-
 mutant enzyme L79A, using sebacconitrile as a substrate Rhodococcus rhodochrous
5.7
-
 mutant enzyme Y175A, using adiponitrile as a substrate Rhodococcus rhodochrous
16.1
-
 wild type enzyme, using glutaronitrile as a substrate Rhodococcus rhodochrous
17.7
-
 mutant enzyme K9A, using glutaronitrile as a substrate Rhodococcus rhodochrous
22.7
-
 mutant enzyme L79A, using glutaronitrile as a substrate Rhodococcus rhodochrous
29.2
-
 mutant enzyme L79A, using adiponitrile as a substrate Rhodococcus rhodochrous
31.4
-
 wild type enzyme, using adiponitrile as a substrate Rhodococcus rhodochrous
40.8
-
 mutant enzyme K9A, using adiponitrile as a substrate Rhodococcus rhodochrous

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2-cyanopyridine + 2 H2O
-
 Rhodococcus rhodochrous 2-pyridine carboxylic acid + NH3
-
 ?
adiponitrile + H2O
-
 Rhodococcus rhodochrous adipic acid + NH3
-
 ?
adiponitrile + H2O
-
 Rhodococcus rhodochrous adipate + NH3
-
 ?
benzonitrile + 2 H2O
-
 Rhodococcus rhodochrous benzoic acid + NH3
-
 ?
glutaronitrile + H2O
-
 Rhodococcus rhodochrous glutarate + NH3
-
 ?
glutaronitrile + H2O
-
 Rhodococcus rhodochrous glutaric acid + NH3
-
 ?
m-tolunitrile + H2O
-
 Rhodococcus rhodochrous m-methylbenzoate + NH3
-
 ?
sebaconitrile + H2O
-
 Rhodococcus rhodochrous ? + NH3
-
 ?
succinonitrile + H2O
-
 Rhodococcus rhodochrous succinate + NH3
-
 ?

Subunits

Subunits Comment Organism
monomer 1 * 40000, SDS-PAGE Rhodococcus rhodochrous

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.0001
-
 m-tolunitrile mutant Y142V, pH 7.5, 25°C Rhodococcus rhodochrous
0.00012
-
 2-Cyanopyridine mutant Y142F, pH 7.5, 25°C Rhodococcus rhodochrous
0.00014
-
 m-tolunitrile mutant Y142F, pH 7.5, 25°C Rhodococcus rhodochrous
0.00014
-
 Benzonitrile mutant Y142V, pH 7.5, 25°C Rhodococcus rhodochrous
0.00015
-
 Benzonitrile wild-type, pH 7.5, 25°C Rhodococcus rhodochrous
0.00015
-
 Benzonitrile mutant K9A, pH 7.5, 25°C Rhodococcus rhodochrous
0.00017
-
 m-tolunitrile mutant K9A, pH 7.5, 25°C Rhodococcus rhodochrous
0.000175
-
 2-Cyanopyridine wild-type, pH 7.5, 25°C Rhodococcus rhodochrous
0.00018
-
 adiponitrile mutant Y142F, pH 7.5, 25°C Rhodococcus rhodochrous
0.00018
-
 glutaronitrile mutant Y142F, pH 7.5, 25°C Rhodococcus rhodochrous
0.00019
-
 2-Cyanopyridine mutant K9A, pH 7.5, 25°C Rhodococcus rhodochrous
0.00019
-
 m-tolunitrile mutant Y142A, pH 7.5, 25°C Rhodococcus rhodochrous
0.0002
-
 Benzonitrile mutant L79A, pH 7.5, 25°C Rhodococcus rhodochrous
0.00024
-
 m-tolunitrile wild-type, pH 7.5, 25°C Rhodococcus rhodochrous
0.00025
-
 2-Cyanopyridine mutant L79A, pH 7.5, 25°C Rhodococcus rhodochrous
0.00025
-
 Benzonitrile mutant Y142F, pH 7.5, 25°C Rhodococcus rhodochrous
0.00028
-
 m-tolunitrile mutant L79A, pH 7.5, 25°C Rhodococcus rhodochrous
0.00029
-
 glutaronitrile wild-type, pH 7.5, 25°C Rhodococcus rhodochrous
0.00029
-
 Benzonitrile mutant Y142A, pH 7.5, 25°C Rhodococcus rhodochrous
0.0003
-
 2-Cyanopyridine mutant Y142A, pH 7.5, 25°C Rhodococcus rhodochrous
0.00031
-
 2-Cyanopyridine mutant Y142A, pH 7.5, 25°C Rhodococcus rhodochrous
0.000315
-
 glutaronitrile mutant L79A, pH 7.5, 25°C Rhodococcus rhodochrous
0.00033
-
 glutaronitrile mutant K9A, pH 7.5, 25°C Rhodococcus rhodochrous
0.00038
-
 m-tolunitrile mutant Y142S, pH 7.5, 25°C Rhodococcus rhodochrous
0.00043
-
 Benzonitrile mutant Y142S, pH 7.5, 25°C Rhodococcus rhodochrous
0.00046
-
 adiponitrile wild-type, pH 7.5, 25°C Rhodococcus rhodochrous
0.0005
-
 adiponitrile mutant L79A, pH 7.5, 25°C Rhodococcus rhodochrous
0.00057
-
 adiponitrile mutant K9A, pH 7.5, 25°C Rhodococcus rhodochrous
0.00073
-
 2-Cyanopyridine mutant Y142S, pH 7.5, 25°C Rhodococcus rhodochrous
0.00147
-
 adiponitrile mutant enzyme Y142F Rhodococcus rhodochrous
0.00147
-
 glutaronitrile mutant enzyme Y142F Rhodococcus rhodochrous
0.0038
-
 adiponitrile wild type enzyme Rhodococcus rhodochrous
0.024
-
 glutaronitrile wild type enzyme Rhodococcus rhodochrous