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Literature summary for 3.5.5.1 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

Cloned(Commentary)

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

Protein Variants

Protein Variants Comment Organism
K9A increased specific activities Rhodococcus rhodochrous
L79A increased specific activities Rhodococcus rhodochrous
R129A inactive Rhodococcus rhodochrous
Y142A mutant exhibits slightly higher kcat/Km values for aromatic nitriles and shows no activity toward aliphatic nitriles Rhodococcus rhodochrous
Y142F mutant shows slightly lower kcat/Km values compared to the wild type enzyme Rhodococcus rhodochrous
Y142I low specific activities Rhodococcus rhodochrous
Y142L low specific activities Rhodococcus rhodochrous
Y142S mutant exhibits slightly higher kcat/Km values for aromatic nitriles and shows no activity toward aliphatic nitriles Rhodococcus rhodochrous
Y142V decreased activity Rhodococcus rhodochrous
Y175A increased specific activities Rhodococcus rhodochrous

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.467
-
m-tolunitrile mutant enzyme Y142V Rhodococcus rhodochrous
0.493
-
Benzonitrile mutant enzyme Y142V Rhodococcus rhodochrous
0.612
-
m-tolunitrile mutant enzyme Y142A Rhodococcus rhodochrous
0.819
-
2-Cyanopyridine mutant enzyme Y142A Rhodococcus rhodochrous
1.068
-
Benzonitrile wild type enzyme Rhodococcus rhodochrous
1.187
-
m-tolunitrile mutant enzyme Y142F Rhodococcus rhodochrous
1.485
-
m-tolunitrile mutant enzyme Y142S Rhodococcus rhodochrous
1.53
-
2-Cyanopyridine wild type enzyme Rhodococcus rhodochrous
1.56
-
2-Cyanopyridine mutant enzyme Y142F Rhodococcus rhodochrous
1.581
-
Benzonitrile mutant enzyme Y142A Rhodococcus rhodochrous
1.815
-
Benzonitrile mutant enzyme Y142S Rhodococcus rhodochrous
1.851
-
m-tolunitrile wild type enzyme Rhodococcus rhodochrous
3.449
-
Benzonitrile mutant enzyme Y142F Rhodococcus rhodochrous
3.752
-
2-Cyanopyridine mutant enzyme Y142S Rhodococcus rhodochrous

Molecular Weight [Da]

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

Organism

Organism UniProt Comment Textmining
Rhodococcus rhodochrous
-
strain ATCC 33278
-

Purification (Commentary)

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

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
3.6
-
mutant enzyme Y175A, using benzonitrile as a substrate Rhodococcus rhodochrous
5
-
mutant enzyme Y175A, using m-tolunitrile as a substrate Rhodococcus rhodochrous
7
-
wild type enzyme, using benzonitrile as a substrate Rhodococcus rhodochrous
7.4
-
mutant enzyme K9A, using benzonitrile as a substrate Rhodococcus rhodochrous
9.2
-
wild type enzyme, using 2-cyanopyridine as a substrate Rhodococcus rhodochrous
10.7
-
wild type enzyme, using m-tolunitrile as a substrate Rhodococcus rhodochrous
11.6
-
mutant enzyme K9A, using 2-cyanopyridine as a substrate Rhodococcus rhodochrous
13.5
-
mutant enzyme Y142A, using 2-cyanopyridine as a substrate Rhodococcus rhodochrous
18.2
-
mutant enzyme K9A, using m-tolunitrile as a substrate Rhodococcus rhodochrous
19.8
-
mutant enzyme L79A, using benzonitrile as a substrate Rhodococcus rhodochrous
22.4
-
mutant enzyme Y142A, using benzonitrile as a substrate Rhodococcus rhodochrous
25
-
mutant enzyme L79A, using m-tolunitrile as a substrate Rhodococcus rhodochrous
26
-
mutant enzyme Y142A, using m-tolunitrile as a substrate Rhodococcus rhodochrous
29
-
mutant enzyme L79A, using 2-cyanopyridine 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
-
?
benzonitrile + 2 H2O
-
Rhodococcus rhodochrous benzoic acid + NH3
-
?
m-tolunitrile + H2O
-
Rhodococcus rhodochrous m-methylbenzoate + 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
0.00085
-
m-tolunitrile mutant enzyme Y142V Rhodococcus rhodochrous
0.00098
-
2-Cyanopyridine mutant enzyme Y142F Rhodococcus rhodochrous
0.00115
-
Benzonitrile mutant enzyme Y142V Rhodococcus rhodochrous
0.00119
-
m-tolunitrile mutant enzyme Y142F Rhodococcus rhodochrous
0.00131
-
Benzonitrile wild type enzyme Rhodococcus rhodochrous
0.00146
-
2-Cyanopyridine wild type enzyme Rhodococcus rhodochrous
0.00157
-
m-tolunitrile mutant enzyme Y142A Rhodococcus rhodochrous
0.00193
-
2-Cyanopyridine mutant enzyme Y142A Rhodococcus rhodochrous
0.00198
-
m-tolunitrile wild type enzyme Rhodococcus rhodochrous
0.0021
-
Benzonitrile mutant enzyme Y142F Rhodococcus rhodochrous
0.0024
-
Benzonitrile mutant enzyme Y142A Rhodococcus rhodochrous
0.00313
-
m-tolunitrile mutant enzyme Y142S Rhodococcus rhodochrous
0.00358
-
Benzonitrile mutant enzyme Y142S Rhodococcus rhodochrous
0.00612
-
2-Cyanopyridine mutant enzyme Y142S Rhodococcus rhodochrous