Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli strain ER2566 | Rhodococcus rhodochrous |
Protein Variants | Comment | Organism |
---|---|---|
G103A | the mutant exhibits decreased specific activity with 3-methylbenzonitrile and increased specific activity with benzonitrile compared to the wild type enzyme | Rhodococcus rhodochrous |
I104A | the mutant exhibits decreased specific activity with 3-methylbenzonitrile and increased specific activity with benzonitrile compared to the wild type enzyme | Rhodococcus rhodochrous |
M114A | the mutant exhibits decreased specific activity with 3-methylbenzonitrile and increased specific activity with benzonitrile compared to the wild type enzyme | Rhodococcus rhodochrous |
N164A | the mutant exhibits decreased specific activity with 3-methylbenzonitrile and increased specific activity with benzonitrile compared to the wild type enzyme | Rhodococcus rhodochrous |
Q116A | the mutant exhibits decreased specific activity with 3-methylbenzonitrile and increased specific activity with benzonitrile compared to the wild type enzyme | Rhodococcus rhodochrous |
R129A | the mutant exhibits no activity with 3-methylbenzonitrile and benzonitrile | Rhodococcus rhodochrous |
R129H | the mutant enzyme is active only for meta-substituted benzonitriles | Rhodococcus rhodochrous |
R129K | the mutant enzyme is active only for meta-substituted benzonitriles | Rhodococcus rhodochrous |
R130A | the mutant exhibits increased specific activity with 3-methylbenzonitrile and benzonitrile compared to the wild type enzyme | Rhodococcus rhodochrous |
S188A | the mutant exhibits decreased specific activity with 3-methylbenzonitrile and increased specific activity with benzonitrile compared to the wild type enzyme | Rhodococcus rhodochrous |
T115A | the mutant exhibits decreased specific activity with 3-methylbenzonitrile and increased specific activity with benzonitrile compared to the wild type enzyme | Rhodococcus rhodochrous |
V49A | the mutant exhibits decreased specific activity with 3-methylbenzonitrile and increased specific activity with benzonitrile compared to the wild type enzyme | Rhodococcus rhodochrous |
Y142A | the mutant exhibits decreased specific activity with 3-methylbenzonitrile and increased specific activity with benzonitrile compared to the wild type enzyme | Rhodococcus rhodochrous |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.456 | - |
3-aminobenzonitrile | mutant enzyme R129K, in 50 mM potassium phosphate (pH 7.5), at 25°C | Rhodococcus rhodochrous | |
0.483 | - |
3-aminobenzonitrile | mutant enzyme R129H, in 50 mM potassium phosphate (pH 7.5), at 25°C | Rhodococcus rhodochrous | |
0.55 | - |
3-methylbenzonitrile | mutant enzyme R129K, in 50 mM potassium phosphate (pH 7.5), at 25°C | Rhodococcus rhodochrous | |
0.592 | - |
3-methylbenzonitrile | mutant enzyme R129H, in 50 mM potassium phosphate (pH 7.5), at 25°C | Rhodococcus rhodochrous | |
1.068 | - |
Benzonitrile | wild type enzyme, in 50 mM potassium phosphate (pH 7.5), at 25°C | Rhodococcus rhodochrous | |
1.468 | - |
Benzonitrile | mutant enzyme R129K, in 50 mM potassium phosphate (pH 7.5), at 25°C | Rhodococcus rhodochrous | |
1.699 | - |
4-aminobenzonitrile | wild type enzyme, in 50 mM potassium phosphate (pH 7.5), at 25°C | Rhodococcus rhodochrous | |
1.851 | - |
3-methylbenzonitrile | wild type enzyme, in 50 mM potassium phosphate (pH 7.5), at 25°C | Rhodococcus rhodochrous | |
1.863 | - |
3-aminobenzonitrile | wild type enzyme, in 50 mM potassium phosphate (pH 7.5), at 25°C | Rhodococcus rhodochrous | |
2.426 | - |
4-methylbenzonitrile | wild type enzyme, in 50 mM potassium phosphate (pH 7.5), at 25°C | Rhodococcus rhodochrous | |
3.13 | - |
Benzonitrile | mutant enzyme R129H, in 50 mM potassium phosphate (pH 7.5), at 25°C | Rhodococcus rhodochrous |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Rhodococcus rhodochrous | - |
- |
- |
Rhodococcus rhodochrous ATCC 33278 | - |
- |
- |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
0.6 | - |
mutant enzyme N164A, using 3-methylbenzonitrile as substrate, in 50 mM potassium phosphate (pH 7.5), at 25°C | Rhodococcus rhodochrous |
5.1 | - |
mutant enzyme V49A, using 3-methylbenzonitrile as substrate, in 50 mM potassium phosphate (pH 7.5), at 25°C | Rhodococcus rhodochrous |
5.2 | - |
mutant enzyme M114A, using 3-methylbenzonitrile as substrate, in 50 mM potassium phosphate (pH 7.5), at 25°C | Rhodococcus rhodochrous |
7 | - |
wild type enzyme, using benzonitrile as substrate, in 50 mM potassium phosphate (pH 7.5), at 25°C | Rhodococcus rhodochrous |
7.5 | - |
mutant enzyme T115A, using 3-methylbenzonitrile as substrate, in 50 mM potassium phosphate (pH 7.5), at 25°C | Rhodococcus rhodochrous |
8 | - |
mutant enzyme S188A, using 3-methylbenzonitrile as substrate, in 50 mM potassium phosphate (pH 7.5), at 25°C | Rhodococcus rhodochrous |
8.2 | - |
mutant enzyme T115A, using benzonitrile as substrate, in 50 mM potassium phosphate (pH 7.5), at 25°C | Rhodococcus rhodochrous |
8.8 | - |
mutant enzyme G103A, using benzonitrile as substrate, in 50 mM potassium phosphate (pH 7.5), at 25°C | Rhodococcus rhodochrous |
8.8 | - |
mutant enzyme I104A, using 3-methylbenzonitrile as substrate, in 50 mM potassium phosphate (pH 7.5), at 25°C | Rhodococcus rhodochrous |
9 | - |
mutant enzyme Q116A, using benzonitrile as substrate, in 50 mM potassium phosphate (pH 7.5), at 25°C | Rhodococcus rhodochrous |
9.6 | - |
mutant enzyme G103A, using 3-methylbenzonitrile as substrate, in 50 mM potassium phosphate (pH 7.5), at 25°C | Rhodococcus rhodochrous |
9.7 | - |
mutant enzyme N164A, using benzonitrile as substrate, in 50 mM potassium phosphate (pH 7.5), at 25°C | Rhodococcus rhodochrous |
9.9 | - |
mutant enzyme M114A, using benzonitrile as substrate, in 50 mM potassium phosphate (pH 7.5), at 25°C | Rhodococcus rhodochrous |
9.9 | - |
mutant enzyme Q116A, using 3-methylbenzonitrile as substrate, in 50 mM potassium phosphate (pH 7.5), at 25°C | Rhodococcus rhodochrous |
10 | - |
mutant enzyme S188A, using benzonitrile as substrate, in 50 mM potassium phosphate (pH 7.5), at 25°C | Rhodococcus rhodochrous |
10.4 | - |
mutant enzyme I104A, using benzonitrile as substrate, in 50 mM potassium phosphate (pH 7.5), at 25°C | Rhodococcus rhodochrous |
10.7 | - |
wild type enzyme, using 3-methylbenzonitrile as substrate, in 50 mM potassium phosphate (pH 7.5), at 25°C | Rhodococcus rhodochrous |
11.6 | - |
mutant enzymeV49A, using benzonitrile as substrate, in 50 mM potassium phosphate (pH 7.5), at 25°C | Rhodococcus rhodochrous |
14.5 | - |
mutant enzyme R130A, using 3-methylbenzonitrile as substrate, in 50 mM potassium phosphate (pH 7.5), at 25°C | Rhodococcus rhodochrous |
17.7 | - |
mutant enzyme R130A, using benzonitrile as substrate, in 50 mM potassium phosphate (pH 7.5), at 25°C | Rhodococcus rhodochrous |
22.4 | - |
mutant enzyme Y142A, using benzonitrile as substrate, in 50 mM potassium phosphate (pH 7.5), at 25°C | Rhodococcus rhodochrous |
26 | - |
mutant enzyme Y142A, using 3-methylbenzonitrile as substrate, in 50 mM potassium phosphate (pH 7.5), at 25°C | Rhodococcus rhodochrous |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
3-aminobenzonitrile + H2O | - |
Rhodococcus rhodochrous | 3-aminobenzoic acid + NH3 | - |
? | |
3-aminobenzonitrile + H2O | - |
Rhodococcus rhodochrous ATCC 33278 | 3-aminobenzoic acid + NH3 | - |
? | |
3-methylbenzonitrile + H2O | - |
Rhodococcus rhodochrous | 3-methylbenzoic acid + NH3 | - |
? | |
3-methylbenzonitrile + H2O | - |
Rhodococcus rhodochrous ATCC 33278 | 3-methylbenzoic acid + NH3 | - |
? | |
4-aminobenzonitrile + H2O | - |
Rhodococcus rhodochrous | 4-aminobenzoic acid + NH3 | - |
? | |
4-aminobenzonitrile + H2O | - |
Rhodococcus rhodochrous ATCC 33278 | 4-aminobenzoic acid + NH3 | - |
? | |
4-methylbenzonitrile + H2O | - |
Rhodococcus rhodochrous | 4-methylbenzoic acid + NH3 | - |
? | |
benzonitrile + 2 H2O | - |
Rhodococcus rhodochrous | benzoic acid + NH3 | - |
? | |
benzonitrile + 2 H2O | - |
Rhodococcus rhodochrous ATCC 33278 | benzoic acid + NH3 | - |
? | |
additional information | the wild type enzyme containing Arg129 is active only for meta- and para-substituted benzonitriles with a methyl or amino group, but shows no activity for ortho-substituted benzonitriles | Rhodococcus rhodochrous | ? | - |
? | |
additional information | the wild type enzyme containing Arg129 is active only for meta- and para-substituted benzonitriles with a methyl or amino group, but shows no activity for ortho-substituted benzonitriles | Rhodococcus rhodochrous ATCC 33278 | ? | - |
? |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.000129 | - |
Benzonitrile | wild type enzyme, in 50 mM potassium phosphate (pH 7.5), at 25°C | Rhodococcus rhodochrous | |
0.00014 | - |
3-aminobenzonitrile | mutant enzyme R129K, in 50 mM potassium phosphate (pH 7.5), at 25°C | Rhodococcus rhodochrous | |
0.00015 | - |
3-aminobenzonitrile | mutant enzyme R129H, in 50 mM potassium phosphate (pH 7.5), at 25°C | Rhodococcus rhodochrous | |
0.000158 | - |
3-methylbenzonitrile | mutant enzyme R129K, in 50 mM potassium phosphate (pH 7.5), at 25°C | Rhodococcus rhodochrous | |
0.000162 | - |
Benzonitrile | mutant enzyme R129K, in 50 mM potassium phosphate (pH 7.5), at 25°C | Rhodococcus rhodochrous | |
0.00018 | - |
3-methylbenzonitrile | mutant enzyme R129H, in 50 mM potassium phosphate (pH 7.5), at 25°C | Rhodococcus rhodochrous | |
0.000198 | - |
3-methylbenzonitrile | wild type enzyme, in 50 mM potassium phosphate (pH 7.5), at 25°C | Rhodococcus rhodochrous | |
0.000283 | - |
4-aminobenzonitrile | wild type enzyme, in 50 mM potassium phosphate (pH 7.5), at 25°C | Rhodococcus rhodochrous | |
0.000288 | - |
3-aminobenzonitrile | wild type enzyme, in 50 mM potassium phosphate (pH 7.5), at 25°C | Rhodococcus rhodochrous | |
0.00047 | - |
Benzonitrile | mutant enzyme R129H, in 50 mM potassium phosphate (pH 7.5), at 25°C | Rhodococcus rhodochrous | |
0.000695 | - |
4-methylbenzonitrile | wild type enzyme, in 50 mM potassium phosphate (pH 7.5), at 25°C | Rhodococcus rhodochrous |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.001068 | - |
3-methylbenzonitrile | wild type enzyme, in 50 mM potassium phosphate (pH 7.5), at 25°C | Rhodococcus rhodochrous | |
0.0011 | - |
Benzonitrile | mutant enzyme R129K, in 50 mM potassium phosphate (pH 7.5), at 25°C | Rhodococcus rhodochrous | |
0.001223 | - |
Benzonitrile | wild type enzyme, in 50 mM potassium phosphate (pH 7.5), at 25°C | Rhodococcus rhodochrous | |
0.0015 | - |
Benzonitrile | mutant enzyme R129H, in 50 mM potassium phosphate (pH 7.5), at 25°C | Rhodococcus rhodochrous | |
0.001548 | - |
3-aminobenzonitrile | wild type enzyme, in 50 mM potassium phosphate (pH 7.5), at 25°C | Rhodococcus rhodochrous | |
0.001667 | - |
4-aminobenzonitrile | wild type enzyme, in 50 mM potassium phosphate (pH 7.5), at 25°C | Rhodococcus rhodochrous | |
0.002867 | - |
4-methylbenzonitrile | wild type enzyme, in 50 mM potassium phosphate (pH 7.5), at 25°C | Rhodococcus rhodochrous | |
0.0029 | - |
3-methylbenzonitrile | mutant enzyme R129K, in 50 mM potassium phosphate (pH 7.5), at 25°C | Rhodococcus rhodochrous | |
0.003 | - |
3-methylbenzonitrile | mutant enzyme R129H, in 50 mM potassium phosphate (pH 7.5), at 25°C | Rhodococcus rhodochrous | |
0.00308 | - |
3-aminobenzonitrile | mutant enzyme R129K, in 50 mM potassium phosphate (pH 7.5), at 25°C | Rhodococcus rhodochrous | |
0.0032 | - |
3-aminobenzonitrile | mutant enzyme R129H, in 50 mM potassium phosphate (pH 7.5), at 25°C | Rhodococcus rhodochrous |