Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli strain ER2566 | Rhodococcus rhodochrous |
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 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 Maximum [Da] | Comment | Organism |
---|---|---|---|
40000 | - |
SDS-PAGE | Rhodococcus rhodochrous |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Rhodococcus rhodochrous | - |
strain ATCC 33278 | - |
Purification (Comment) | Organism |
---|---|
HisTrap HP 9 column chromatography | Rhodococcus rhodochrous |
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 | 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 | Comment | Organism |
---|---|---|
monomer | 1 * 40000, SDS-PAGE | Rhodococcus rhodochrous |
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 |