Cloned (Comment) | Organism |
---|---|
recombinant expression of wild-type and mutant enzymes in Escherichia coli strain BL21(DE3) | Brevundimonas diminuta |
Crystallization (Comment) | Organism |
---|---|
purified recombinant enzyme mutants A80V/F132V/K185R/H254Q/H257Y/I274N/S308L and I106C/F132V/H254Q/H257Y/A270V/L272M/I274N/S308L, vapor diffusion method, mixing of 0.001 ml of 10 mg/ml protein in 50 mM HEPES, pH 8.5, and 1.0 mM CoCl2, with 0.001 ml of precipitant solution containing 100 mM sodium cacodylate, pH 5.5-7.0, 0.2 M magnesium acetate, and 15-30% PEG 8000, and equilbration against 0.5 ml of the precipitant solution, 7-21 days, 18°C, X-ray diffraction structure determination and analysis, molecular replacement using the coordinates of wild-type PTE, PDB ID 1DPM, as the search model | Brevundimonas diminuta |
Protein Variants | Comment | Organism |
---|---|---|
A80V/F132V/K185R/H254Q/H257Y/I274N | site-directed mutagenesis | Brevundimonas diminuta |
A80V/F132V/K185R/H254Q/H257Y/I274N/S308L | site-directed mutagenesis, the mutant combines expression-enhancing mutations (A80V/K185R/I274N) with additional changes in the active site to achieve a high kcat/Km for the hydrolysis of the SP-enantiomer of ethyl-S-(2-diisopropylaminoethyl)methylphosphonothiolate, i.e. VX. Three-dimensional structure, overview | Brevundimonas diminuta |
A80V/I106C/F132V/K185R/H254Q/H257Y/I274N | site-directed mutagenesis | Brevundimonas diminuta |
A80V/I106C/F132V/K185R/H254Q/H257Y/I274N/S308L | site-directed mutagenesis | Brevundimonas diminuta |
A80V/I106G/F132V/K185R/H254Q/H257Y/I274N | site-directed mutagenesis | Brevundimonas diminuta |
A80V/I106G/F132V/K185R/H254Q/H257Y/I274N/S308L | site-directed mutagenesis | Brevundimonas diminuta |
F132V/H254Q/H257F | site-directed mutagenesis | Brevundimonas diminuta |
F132V/H254Q/H257F/S308L | site-directed mutagenesis | Brevundimonas diminuta |
H254Q/H257F | site-directed mutagenesis, the mutant exhibits a 100fold improvement for the hydrolysis of O-ethyl-S-(2-diisopropylaminoethyl)methylphosphonothiolate, i.e. VX, relative to the wild-type enzyme | Brevundimonas diminuta |
I106C | site-directed mutagenesis | Brevundimonas diminuta |
I106C/F132V/H254Q/H257F | site-directed mutagenesis | Brevundimonas diminuta |
I106C/F132V/H254Q/H257F/S308L | site-directed mutagenesis, the mutant shows a similar catalytic efficiency for the hydrolysis of ethyl-S-(2-diisopropylaminoethyl)methylphosphonothiolate, i.e. VX, as mutant H254Q/H257F, but a 3fold improvement in kcat | Brevundimonas diminuta |
I106C/F132V/H254Q/H257Y/A270V/L272M/I274N/S308L | site-directed mutagenesis, the mutant has a kcat value for ethyl-S-(2-diisopropylaminoethyl)methylphosphonothiolate, i.e. VX, that is enhanced by 150fold relative to wild-type PTE. Three-dimensional structure, overview | Brevundimonas diminuta |
I106C/F132V/H254Q/H257Y/A270V/L272M/I274N/S308L/Y309F | site-directed mutagenesis | Brevundimonas diminuta |
I106G | site-directed mutagenesis | Brevundimonas diminuta |
I106G/F132V/H254Q/H257F/S308L | site-directed mutagenesis | Brevundimonas diminuta |
I106G/F132V/H254Q/H257Y/A270V/L272M/I274N/S308L | site-directed mutagenesis | Brevundimonas diminuta |
L308S | site-directed mutagenesis | Brevundimonas diminuta |
additional information | computational docking study of wild-type and mutant enzymes with the toxic substrates | Brevundimonas diminuta |
Y309F | site-directed mutagenesis | Brevundimonas diminuta |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0017 | - |
diethyl-paraoxon | pH 9.0, 30°C, recombinant mutant A80V/F132V/K185R/H254Q/H257Y/I274N | Brevundimonas diminuta | |
0.0036 | - |
diethyl-paraoxon | pH 9.0, 30°C, recombinant mutant A80V/I106C/F132V/K185R/H254Q/H257Y/I274N | Brevundimonas diminuta | |
0.0053 | - |
diethyl-paraoxon | pH 9.0, 30°C, recombinant mutant A80V/I106C/F132V/K185R/H254Q/H257Y/I274N/S308L | Brevundimonas diminuta | |
0.0053 | - |
diethyl-paraoxon | pH 9.0, 30°C, recombinant mutant H254Q/H257F | Brevundimonas diminuta | |
0.0054 | - |
diethyl-paraoxon | pH 9.0, 30°C, recombinant mutant F132V/H254Q/H257F/S308L | Brevundimonas diminuta | |
0.0056 | - |
diethyl-paraoxon | pH 9.0, 30°C, recombinant mutant I106C/F132V/H254Q/H257F/S308L | Brevundimonas diminuta | |
0.0067 | - |
diethyl-paraoxon | pH 9.0, 30°C, recombinant mutant I106C/F132V/H254Q/H257F | Brevundimonas diminuta | |
0.0072 | - |
diethyl-paraoxon | pH 9.0, 30°C, recombinant mutant I106C/F132V/H254Q/H257Y/A270V/L272M/I274N/S308L | Brevundimonas diminuta | |
0.008 | - |
diethyl-paraoxon | pH 9.0, 30°C, recombinant mutant A80V/F132V/K185R/H254Q/H257Y/I274N/S308L | Brevundimonas diminuta | |
0.0092 | - |
diethyl-paraoxon | pH 9.0, 30°C, recombinant mutant A80V/I106G/F132V/K185R/H254Q/H257Y/I274N | Brevundimonas diminuta | |
0.0092 | - |
diethyl-paraoxon | pH 9.0, 30°C, recombinant mutant I106C/F132V/H254Q/H257Y/A270V/L272M/I274N/S308L/Y309F | Brevundimonas diminuta | |
0.011 | - |
diethyl-paraoxon | pH 9.0, 30°C, recombinant mutant F132V/H254Q/H257F | Brevundimonas diminuta | |
0.021 | - |
diethyl-paraoxon | pH 9.0, 30°C, recombinant mutant A80V/I106G/F132V/K185R/H254Q/H257Y/I274N/S308L | Brevundimonas diminuta | |
0.022 | - |
diethyl-paraoxon | pH 9.0, 30°C, recombinant mutant I106G/F132V/H254Q/H257F/S308L | Brevundimonas diminuta | |
0.025 | - |
diethyl-paraoxon | pH 9.0, 30°C, recombinant mutant I106G/F132V/H254Q/H257F | Brevundimonas diminuta | |
0.03 | - |
4-acetylphenyl (S)-2-methylpropyl methylphosphonate | pH 9.0, 30°C, recombinant mutant H254Q/H257F | Brevundimonas diminuta | |
0.033 | - |
diethyl-paraoxon | pH 9.0, 30°C, recombinant mutant I106G/F132V/H254Q/H257Y/A270V/L272M/I274N/S308L | Brevundimonas diminuta | |
0.081 | - |
diethyl-paraoxon | pH 9.0, 30°C, recombinant wild-type enzyme | Brevundimonas diminuta | |
0.13 | - |
4-acetylphenyl (S)-2-methylpropyl methylphosphonate | pH 9.0, 30°C, recombinant mutant A80V/I106C/F132V/K185R/H254Q/H257Y/I274N | Brevundimonas diminuta | |
0.15 | - |
4-acetylphenyl (R)-2-methylpropyl methylphosphonate | pH 9.0, 30°C, recombinant mutant F132V/H254Q/H257F/S308L | Brevundimonas diminuta | |
0.16 | - |
4-acetylphenyl (R)-2-methylpropyl methylphosphonate | pH 9.0, 30°C, recombinant mutant A80V/F132V/K185R/H254Q/H257Y/I274N/S308L | Brevundimonas diminuta | |
0.18 | - |
4-acetylphenyl (S)-2-methylpropyl methylphosphonate | pH 9.0, 30°C, recombinant mutant A80V/I106C/F132V/K185R/H254Q/H257Y/I274N | Brevundimonas diminuta | |
0.18 | - |
4-acetylphenyl (S)-2-methylpropyl methylphosphonate | pH 9.0, 30°C, recombinant mutant A80V/I106G/F132V/K185R/H254Q/H257Y/I274N/S308L | Brevundimonas diminuta | |
0.18 | - |
4-acetylphenyl (R)-2-methylpropyl methylphosphonate | pH 9.0, 30°C, recombinant mutant I106C/F132V/H254Q/H257F/S308L | Brevundimonas diminuta | |
0.19 | - |
4-acetylphenyl (S)-2-methylpropyl methylphosphonate | pH 9.0, 30°C, recombinant mutant I106C/F132V/H254Q/H257F | Brevundimonas diminuta | |
0.19 | - |
4-acetylphenyl (S)-2-methylpropyl methylphosphonate | pH 9.0, 30°C, recombinant mutant I106C/F132V/H254Q/H257F/S308L | Brevundimonas diminuta | |
0.2 | - |
4-acetylphenyl (S)-2-methylpropyl methylphosphonate | pH 9.0, 30°C, recombinant mutant I106G/F132V/H254Q/H257F/S308L | Brevundimonas diminuta | |
0.23 | - |
4-acetylphenyl (R)-2-methylpropyl methylphosphonate | pH 9.0, 30°C, recombinant mutant A80V/I106C/F132V/K185R/H254Q/H257Y/I274N/S308L | Brevundimonas diminuta | |
0.23 | - |
4-acetylphenyl (S)-2-methylpropyl methylphosphonate | pH 9.0, 30°C, recombinant mutant A80V/I106C/F132V/K185R/H254Q/H257Y/I274N/S308L | Brevundimonas diminuta | |
0.24 | - |
4-acetylphenyl (S)-2-methylpropyl methylphosphonate | pH 9.0, 30°C, recombinant mutant I106G/F132V/H254Q/H257F | Brevundimonas diminuta | |
0.31 | - |
4-acetylphenyl (S)-2-methylpropyl methylphosphonate | pH 9.0, 30°C, recombinant mutant I106G/F132V/H254Q/H257Y/A270V/L272M/I274N/S308L | Brevundimonas diminuta | |
0.35 | - |
(S)-[2-[di(propan-2-yl)amino]ethyl] O,O-diethyl phosphorothioate | pH 9.0, 30°C, recombinant mutant A80V/F132V/K185R/H254Q/H257Y/I274N | Brevundimonas diminuta | |
0.36 | - |
4-acetylphenyl (R)-2-methylpropyl methylphosphonate | pH 9.0, 30°C, recombinant mutant H254Q/H257F | Brevundimonas diminuta | |
0.41 | - |
4-acetylphenyl (R)-2-methylpropyl methylphosphonate | pH 9.0, 30°C, recombinant mutant A80V/I106G/F132V/K185R/H254Q/H257Y/I274N/S308L | Brevundimonas diminuta | |
0.5 | - |
4-acetylphenyl (S)-2-methylpropyl methylphosphonate | pH 9.0, 30°C, recombinant mutant I106C/F132V/H254Q/H257Y/A270V/L272M/I274N/S308L | Brevundimonas diminuta | |
0.6 | - |
(S)-[2-[di(propan-2-yl)amino]ethyl] O,O-diethyl phosphorothioate | pH 9.0, 30°C, recombinant mutant I106C/F132V/H254Q/H257Y/A270V/L272M/I274N/S308L | Brevundimonas diminuta | |
0.62 | - |
4-acetylphenyl (R)-2-methylpropyl methylphosphonate | pH 9.0, 30°C, recombinant mutant I106C/F132V/H254Q/H257Y/A270V/L272M/I274N/S308L | Brevundimonas diminuta | |
0.65 | - |
(S)-[2-[di(propan-2-yl)amino]ethyl] O,O-diethyl phosphorothioate | pH 9.0, 30°C, recombinant mutant F132V/H254Q/H257F/S308L | Brevundimonas diminuta | |
0.71 | - |
4-acetylphenyl (R)-2-methylpropyl methylphosphonate | pH 9.0, 30°C, recombinant mutant I106G/F132V/H254Q/H257Y/A270V/L272M/I274N/S308L | Brevundimonas diminuta | |
0.73 | - |
(S)-[2-[di(propan-2-yl)amino]ethyl] O,O-diethyl phosphorothioate | pH 9.0, 30°C, recombinant mutant A80V/F132V/K185R/H254Q/H257Y/I274N/S308L | Brevundimonas diminuta | |
0.76 | - |
(S)-[2-[di(propan-2-yl)amino]ethyl] O,O-diethyl phosphorothioate | pH 9.0, 30°C, recombinant mutant I106C/F132V/H254Q/H257F/S308L | Brevundimonas diminuta | |
0.8 | - |
(S)-[2-[di(propan-2-yl)amino]ethyl] O,O-diethyl phosphorothioate | pH 9.0, 30°C, recombinant mutant A80V/I106C/F132V/K185R/H254Q/H257Y/I274N/S308L | Brevundimonas diminuta | |
0.87 | - |
(S)-[2-[di(propan-2-yl)amino]ethyl] O,O-diethyl phosphorothioate | pH 9.0, 30°C, recombinant wild-type enzyme | Brevundimonas diminuta | |
1 | - |
(S)-[2-[di(propan-2-yl)amino]ethyl] O,O-diethyl phosphorothioate | pH 9.0, 30°C, recombinant mutant F132V/H254Q/H257F | Brevundimonas diminuta | |
1.2 | - |
(S)-[2-[di(propan-2-yl)amino]ethyl] O,O-diethyl phosphorothioate | pH 9.0, 30°C, recombinant mutant I106C/F132V/H254Q/H257Y/A270V/L272M/I274N/S308L/Y309F | Brevundimonas diminuta | |
1.4 | - |
(S)-[2-[di(propan-2-yl)amino]ethyl] O,O-diethyl phosphorothioate | pH 9.0, 30°C, recombinant mutant H254Q/H257F | Brevundimonas diminuta | |
1.4 | - |
4-acetylphenyl (R)-2-methylpropyl methylphosphonate | pH 9.0, 30°C, recombinant mutant I106G/F132V/H254Q/H257F/S308L | Brevundimonas diminuta | |
1.5 | - |
4-acetylphenyl (S)-2-methylpropyl methylphosphonate | pH 9.0, 30°C, recombinant mutant A80V/F132V/K185R/H254Q/H257Y/I274N | Brevundimonas diminuta | |
1.5 | - |
4-acetylphenyl (S)-2-methylpropyl methylphosphonate | pH 9.0, 30°C, recombinant mutant F132V/H254Q/H257F/S308L | Brevundimonas diminuta | |
1.6 | - |
(S)-[2-[di(propan-2-yl)amino]ethyl] O,O-diethyl phosphorothioate | pH 9.0, 30°C, recombinant mutant A80V/I106C/F132V/K185R/H254Q/H257Y/I274N | Brevundimonas diminuta | |
1.7 | - |
4-acetylphenyl (R)-2-methylpropyl methylphosphonate | pH 9.0, 30°C, recombinant wild-type enzyme | Brevundimonas diminuta | |
1.7 | - |
4-acetylphenyl (R)-2-methylpropyl methylphosphonate | pH 9.0, 30°C, recombinant mutant A80V/F132V/K185R/H254Q/H257Y/I274N | Brevundimonas diminuta | |
1.7 | - |
4-acetylphenyl (S)-2-methylpropyl methylphosphonate | pH 9.0, 30°C, recombinant mutant A80V/F132V/K185R/H254Q/H257Y/I274N/S308L | Brevundimonas diminuta | |
2 | - |
4-acetylphenyl (R)-2-methylpropyl methylphosphonate | pH 9.0, 30°C, recombinant mutant I106C/F132V/H254Q/H257F | Brevundimonas diminuta | |
2.1 | - |
4-acetylphenyl (R)-2-methylpropyl methylphosphonate | pH 9.0, 30°C, recombinant mutant A80V/I106C/F132V/K185R/H254Q/H257Y/I274N | Brevundimonas diminuta | |
2.8 | - |
(S)-[2-[di(propan-2-yl)amino]ethyl] O,O-diethyl phosphorothioate | pH 9.0, 30°C, recombinant mutant I106G/F132V/H254Q/H257F/S308L | Brevundimonas diminuta | |
3.2 | - |
(S)-[2-[di(propan-2-yl)amino]ethyl] O,O-diethyl phosphorothioate | pH 9.0, 30°C, recombinant mutant I106C/F132V/H254Q/H257F | Brevundimonas diminuta | |
4 | - |
4-acetylphenyl (R)-2-methylpropyl methylphosphonate | pH 9.0, 30°C, recombinant mutant I106G/F132V/H254Q/H257F | Brevundimonas diminuta | |
4.5 | - |
4-acetylphenyl (S)-2-methylpropyl methylphosphonate | pH 9.0, 30°C, recombinant wild-type enzyme | Brevundimonas diminuta |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Co2+ | activates, required | Brevundimonas diminuta |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
diethyl-paraoxon + H2O | Brevundimonas diminuta | - |
diethyl phosphate + 4-nitrophenol | - |
? | |
O-ethyl-S-(2-diisopropylaminoethyl)methylphosphonothiolate + H2O | Brevundimonas diminuta | i.e. VX | ? | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Brevundimonas diminuta | P0A434 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant wild-type and mutant enzymes from cell-free enzyme extract by precipitation of nucleic acids by protamine sulfate, ammonium sulfate fractionation, and gel filtration, followed by anion exchange chromatography | Brevundimonas diminuta |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
(S)-[2-(diethylamino)ethyl] O,O-diethyl phosphorothioate + H2O | - |
Brevundimonas diminuta | diethyl phosphate + 2-(diethylamino)ethane-1-thiol | - |
? | |
(S)-[2-(diethylamino)ethyl] O,O-dimethyl phosphorothioate + H2O | - |
Brevundimonas diminuta | dimethyl phosphate + 2-(diethylamino)ethane-1-thiol | - |
? | |
(S)-[2-(diethylamino)ethyl] O-(2-methylpropyl) methylphosphonothioate + H2O | - |
Brevundimonas diminuta | 2-methylpropyl methylphosphonate + 2-(diethylamino)ethane-1-thiol | - |
? | |
(S)-[2-[di(propan-2-yl)amino]ethyl] O,O-diethyl phosphorothioate + H2O | - |
Brevundimonas diminuta | diethyl phosphate + 2-[di(propan-2-yl)amino]ethane-1-thiol | - |
? | |
(S)-[2-[di(propan-2-yl)amino]ethyl] O,O-dimethyl phosphorothioate + H2O | - |
Brevundimonas diminuta | dimethyl phosphate + 2-[di(propan-2-yl)amino]ethane-1-thiol | - |
? | |
(S)-[2-[di(propan-2-yl)amino]ethyl] O-ethyl methylphosphonothioate + H2O | - |
Brevundimonas diminuta | ethyl methylphosphonate + [di(propan-2-yl)amino]ethane-1-thiol | - |
? | |
4-acetylphenyl (R)-2-methylpropyl methylphosphonate + H2O | - |
Brevundimonas diminuta | (R)-2-methylpropyl methylphosphonate + 4-acetylphenol | - |
? | |
4-acetylphenyl (S)-2-methylpropyl methylphosphonate + H2O | - |
Brevundimonas diminuta | (S)-2-methylpropyl methylphosphonate + 4-acetylphenol | - |
? | |
diethyl-paraoxon + H2O | - |
Brevundimonas diminuta | diethyl phosphate + 4-nitrophenol | - |
? | |
additional information | wild-type PTE exhibits little stereoselectivity against the relatively small phosphonate center of O-ethyl-S-(2-diisopropylaminoethyl)methylphosphonothiolate, i.e. VX, but the elevated pKa of the thiol-leaving group provides a significant challenge for enzyme-catalyzed hydrolysis. Wild-type PTE has enzymatic activity for the hydrolysis of racemic VR similar to VX, but the larger isobutyl group attached to the phosphorus center results in a 25fold preference for the RP-enantiomer. Computational docking study of wild-type and mutant enzymes with the substrates | Brevundimonas diminuta | ? | - |
? | |
N,N-diethyl-2-(methyl-(2-methylpropoxy)phosphoryl)sulfanylethanamine + H2O | i.e. VR | Brevundimonas diminuta | ? | - |
? | |
O-ethyl-S-(2-diisopropylaminoethyl)methylphosphonothiolate + H2O | i.e. VX | Brevundimonas diminuta | ? | - |
? |
Synonyms | Comment | Organism |
---|---|---|
phosphotriesterase | - |
Brevundimonas diminuta |
PTE | - |
Brevundimonas diminuta |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
30 | - |
assay at | Brevundimonas diminuta |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
1.1 | - |
(S)-[2-[di(propan-2-yl)amino]ethyl] O,O-diethyl phosphorothioate | pH 9.0, 30°C, recombinant wild-type enzyme | Brevundimonas diminuta | |
3 | 6 | (S)-[2-[di(propan-2-yl)amino]ethyl] O,O-diethyl phosphorothioate | pH 9.0, 30°C, recombinant mutant I106C/F132V/H254Q/H257Y/A270V/L272M/I274N/S308L/Y309F | Brevundimonas diminuta | |
3 | 6 | 4-acetylphenyl (S)-2-methylpropyl methylphosphonate | pH 9.0, 30°C, recombinant mutant I106G/F132V/H254Q/H257F/S308L | Brevundimonas diminuta | |
5 | - |
4-acetylphenyl (S)-2-methylpropyl methylphosphonate | pH 9.0, 30°C, recombinant mutant A80V/I106C/F132V/K185R/H254Q/H257Y/I274N | Brevundimonas diminuta | |
5.1 | - |
(S)-[2-[di(propan-2-yl)amino]ethyl] O,O-diethyl phosphorothioate | pH 9.0, 30°C, recombinant mutant A80V/F132V/K185R/H254Q/H257Y/I274N | Brevundimonas diminuta | |
6.1 | - |
(S)-[2-[di(propan-2-yl)amino]ethyl] O,O-diethyl phosphorothioate | pH 9.0, 30°C, recombinant mutant H254Q/H257F | Brevundimonas diminuta | |
8.1 | - |
4-acetylphenyl (S)-2-methylpropyl methylphosphonate | pH 9.0, 30°C, recombinant mutant I106C/F132V/H254Q/H257F | Brevundimonas diminuta | |
8.6 | - |
(S)-[2-[di(propan-2-yl)amino]ethyl] O,O-diethyl phosphorothioate | pH 9.0, 30°C, recombinant mutant A80V/I106C/F132V/K185R/H254Q/H257Y/I274N | Brevundimonas diminuta | |
8.7 | - |
4-acetylphenyl (S)-2-methylpropyl methylphosphonate | pH 9.0, 30°C, recombinant mutant H254Q/H257F | Brevundimonas diminuta | |
13 | - |
(S)-[2-[di(propan-2-yl)amino]ethyl] O,O-diethyl phosphorothioate | pH 9.0, 30°C, recombinant mutant I106C/F132V/H254Q/H257F | Brevundimonas diminuta | |
14 | - |
(S)-[2-[di(propan-2-yl)amino]ethyl] O,O-diethyl phosphorothioate | pH 9.0, 30°C, recombinant mutant F132V/H254Q/H257F/S308L | Brevundimonas diminuta | |
14 | - |
4-acetylphenyl (S)-2-methylpropyl methylphosphonate | pH 9.0, 30°C, recombinant mutant F132V/H254Q/H257F/S308L | Brevundimonas diminuta | |
15 | - |
diethyl-paraoxon | pH 9.0, 30°C, recombinant mutant A80V/F132V/K185R/H254Q/H257Y/I274N | Brevundimonas diminuta | |
16 | - |
(S)-[2-[di(propan-2-yl)amino]ethyl] O,O-diethyl phosphorothioate | pH 9.0, 30°C, recombinant mutant I106C/F132V/H254Q/H257F/S308L | Brevundimonas diminuta | |
17 | - |
4-acetylphenyl (R)-2-methylpropyl methylphosphonate | pH 9.0, 30°C, recombinant mutant A80V/I106G/F132V/K185R/H254Q/H257Y/I274N/S308L | Brevundimonas diminuta | |
19 | - |
(S)-[2-[di(propan-2-yl)amino]ethyl] O,O-diethyl phosphorothioate | pH 9.0, 30°C, recombinant mutant F132V/H254Q/H257F | Brevundimonas diminuta | |
21 | - |
4-acetylphenyl (S)-2-methylpropyl methylphosphonate | pH 9.0, 30°C, recombinant mutant I106C/F132V/H254Q/H257F/S308L | Brevundimonas diminuta | |
22 | - |
(S)-[2-[di(propan-2-yl)amino]ethyl] O,O-diethyl phosphorothioate | pH 9.0, 30°C, recombinant mutant A80V/F132V/K185R/H254Q/H257Y/I274N/S308L | Brevundimonas diminuta | |
23 | - |
(S)-[2-[di(propan-2-yl)amino]ethyl] O,O-diethyl phosphorothioate | pH 9.0, 30°C, recombinant mutant A80V/I106C/F132V/K185R/H254Q/H257Y/I274N/S308L | Brevundimonas diminuta | |
25 | - |
4-acetylphenyl (S)-2-methylpropyl methylphosphonate | pH 9.0, 30°C, recombinant wild-type enzyme | Brevundimonas diminuta | |
29 | - |
4-acetylphenyl (S)-2-methylpropyl methylphosphonate | pH 9.0, 30°C, recombinant mutant A80V/F132V/K185R/H254Q/H257Y/I274N/S308L | Brevundimonas diminuta | |
31 | - |
(S)-[2-[di(propan-2-yl)amino]ethyl] O,O-diethyl phosphorothioate | pH 9.0, 30°C, recombinant mutant I106G/F132V/H254Q/H257F/S308L | Brevundimonas diminuta | |
33 | - |
4-acetylphenyl (S)-2-methylpropyl methylphosphonate | pH 9.0, 30°C, recombinant mutant A80V/I106C/F132V/K185R/H254Q/H257Y/I274N/S308L | Brevundimonas diminuta | |
33 | - |
diethyl-paraoxon | pH 9.0, 30°C, recombinant mutant I106C/F132V/H254Q/H257F | Brevundimonas diminuta | |
35 | - |
diethyl-paraoxon | pH 9.0, 30°C, recombinant mutant A80V/I106C/F132V/K185R/H254Q/H257Y/I274N | Brevundimonas diminuta | |
38 | - |
diethyl-paraoxon | pH 9.0, 30°C, recombinant mutant I106C/F132V/H254Q/H257F/S308L | Brevundimonas diminuta | |
40 | - |
4-acetylphenyl (S)-2-methylpropyl methylphosphonate | pH 9.0, 30°C, recombinant mutant I106G/F132V/H254Q/H257F | Brevundimonas diminuta | |
41 | - |
diethyl-paraoxon | pH 9.0, 30°C, recombinant mutant H254Q/H257F | Brevundimonas diminuta | |
45 | - |
4-acetylphenyl (R)-2-methylpropyl methylphosphonate | pH 9.0, 30°C, recombinant mutant A80V/I106C/F132V/K185R/H254Q/H257Y/I274N | Brevundimonas diminuta | |
46 | - |
4-acetylphenyl (R)-2-methylpropyl methylphosphonate | pH 9.0, 30°C, recombinant mutant I106C/F132V/H254Q/H257F/S308L | Brevundimonas diminuta | |
48 | - |
diethyl-paraoxon | pH 9.0, 30°C, recombinant mutant I106G/F132V/H254Q/H257F | Brevundimonas diminuta | |
50 | - |
4-acetylphenyl (R)-2-methylpropyl methylphosphonate | pH 9.0, 30°C, recombinant mutant F132V/H254Q/H257F/S308L | Brevundimonas diminuta | |
51 | - |
4-acetylphenyl (S)-2-methylpropyl methylphosphonate | pH 9.0, 30°C, recombinant mutant A80V/F132V/K185R/H254Q/H257Y/I274N | Brevundimonas diminuta | |
51 | - |
(S)-[2-[di(propan-2-yl)amino]ethyl] O,O-diethyl phosphorothioate | pH 9.0, 30°C, recombinant mutant I106C/F132V/H254Q/H257Y/A270V/L272M/I274N/S308L | Brevundimonas diminuta | |
53 | - |
4-acetylphenyl (S)-2-methylpropyl methylphosphonate | pH 9.0, 30°C, recombinant mutant A80V/I106C/F132V/K185R/H254Q/H257Y/I274N | Brevundimonas diminuta | |
55 | - |
4-acetylphenyl (R)-2-methylpropyl methylphosphonate | pH 9.0, 30°C, recombinant mutant A80V/F132V/K185R/H254Q/H257Y/I274N/S308L | Brevundimonas diminuta | |
56 | - |
4-acetylphenyl (S)-2-methylpropyl methylphosphonate | pH 9.0, 30°C, recombinant mutant I106C/F132V/H254Q/H257Y/A270V/L272M/I274N/S308L | Brevundimonas diminuta | |
57 | - |
4-acetylphenyl (R)-2-methylpropyl methylphosphonate | pH 9.0, 30°C, recombinant mutant H254Q/H257F | Brevundimonas diminuta | |
58 | - |
diethyl-paraoxon | pH 9.0, 30°C, recombinant mutant A80V/I106G/F132V/K185R/H254Q/H257Y/I274N | Brevundimonas diminuta | |
58 | - |
4-acetylphenyl (R)-2-methylpropyl methylphosphonate | pH 9.0, 30°C, recombinant mutant I106G/F132V/H254Q/H257Y/A270V/L272M/I274N/S308L | Brevundimonas diminuta | |
66 | - |
diethyl-paraoxon | pH 9.0, 30°C, recombinant mutant F132V/H254Q/H257F/S308L | Brevundimonas diminuta | |
72 | - |
4-acetylphenyl (R)-2-methylpropyl methylphosphonate | pH 9.0, 30°C, recombinant mutant A80V/I106C/F132V/K185R/H254Q/H257Y/I274N/S308L | Brevundimonas diminuta | |
84 | - |
4-acetylphenyl (R)-2-methylpropyl methylphosphonate | pH 9.0, 30°C, recombinant wild-type enzyme | Brevundimonas diminuta | |
85 | - |
diethyl-paraoxon | pH 9.0, 30°C, recombinant mutant I106C/F132V/H254Q/H257Y/A270V/L272M/I274N/S308L | Brevundimonas diminuta | |
100 | - |
4-acetylphenyl (R)-2-methylpropyl methylphosphonate | pH 9.0, 30°C, recombinant mutant I106G/F132V/H254Q/H257F | Brevundimonas diminuta | |
101 | - |
4-acetylphenyl (R)-2-methylpropyl methylphosphonate | pH 9.0, 30°C, recombinant mutant I106C/F132V/H254Q/H257Y/A270V/L272M/I274N/S308L | Brevundimonas diminuta | |
103 | - |
diethyl-paraoxon | pH 9.0, 30°C, recombinant mutant A80V/I106C/F132V/K185R/H254Q/H257Y/I274N/S308L | Brevundimonas diminuta | |
108 | - |
diethyl-paraoxon | pH 9.0, 30°C, recombinant mutant F132V/H254Q/H257F | Brevundimonas diminuta | |
116 | - |
diethyl-paraoxon | pH 9.0, 30°C, recombinant mutant A80V/F132V/K185R/H254Q/H257Y/I274N/S308L | Brevundimonas diminuta | |
122 | - |
4-acetylphenyl (R)-2-methylpropyl methylphosphonate | pH 9.0, 30°C, recombinant mutant I106C/F132V/H254Q/H257F | Brevundimonas diminuta | |
131 | - |
diethyl-paraoxon | pH 9.0, 30°C, recombinant mutant I106C/F132V/H254Q/H257Y/A270V/L272M/I274N/S308L/Y309F | Brevundimonas diminuta | |
159 | - |
4-acetylphenyl (S)-2-methylpropyl methylphosphonate | pH 9.0, 30°C, recombinant mutant A80V/I106G/F132V/K185R/H254Q/H257Y/I274N/S308L | Brevundimonas diminuta | |
160 | - |
4-acetylphenyl (R)-2-methylpropyl methylphosphonate | pH 9.0, 30°C, recombinant mutant A80V/F132V/K185R/H254Q/H257Y/I274N | Brevundimonas diminuta | |
166 | - |
4-acetylphenyl (S)-2-methylpropyl methylphosphonate | pH 9.0, 30°C, recombinant mutant I106G/F132V/H254Q/H257Y/A270V/L272M/I274N/S308L | Brevundimonas diminuta | |
174 | - |
4-acetylphenyl (R)-2-methylpropyl methylphosphonate | pH 9.0, 30°C, recombinant mutant I106G/F132V/H254Q/H257F/S308L | Brevundimonas diminuta | |
446 | - |
diethyl-paraoxon | pH 9.0, 30°C, recombinant mutant A80V/I106G/F132V/K185R/H254Q/H257Y/I274N/S308L | Brevundimonas diminuta | |
456 | - |
diethyl-paraoxon | pH 9.0, 30°C, recombinant mutant I106G/F132V/H254Q/H257F/S308L | Brevundimonas diminuta | |
545 | - |
diethyl-paraoxon | pH 9.0, 30°C, recombinant mutant I106G/F132V/H254Q/H257Y/A270V/L272M/I274N/S308L | Brevundimonas diminuta | |
2230 | - |
diethyl-paraoxon | pH 9.0, 30°C, recombinant wild-type enzyme | Brevundimonas diminuta |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
8 | 9 | assay at | Brevundimonas diminuta |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
1.264 | - |
(S)-[2-[di(propan-2-yl)amino]ethyl] O,O-diethyl phosphorothioate | pH 9.0, 30°C, recombinant wild-type enzyme | Brevundimonas diminuta | |
4.0625 | - |
(S)-[2-[di(propan-2-yl)amino]ethyl] O,O-diethyl phosphorothioate | pH 9.0, 30°C, recombinant mutant I106C/F132V/H254Q/H257F | Brevundimonas diminuta | |
4.357 | - |
(S)-[2-[di(propan-2-yl)amino]ethyl] O,O-diethyl phosphorothioate | pH 9.0, 30°C, recombinant mutant H254Q/H257F | Brevundimonas diminuta | |
5.375 | - |
(S)-[2-[di(propan-2-yl)amino]ethyl] O,O-diethyl phosphorothioate | pH 9.0, 30°C, recombinant mutant A80V/I106C/F132V/K185R/H254Q/H257Y/I274N | Brevundimonas diminuta | |
5.55 | - |
4-acetylphenyl (S)-2-methylpropyl methylphosphonate | pH 9.0, 30°C, recombinant wild-type enzyme | Brevundimonas diminuta | |
9.3 | - |
4-acetylphenyl (S)-2-methylpropyl methylphosphonate | pH 9.0, 30°C, recombinant mutant F132V/H254Q/H257F/S308L | Brevundimonas diminuta | |
11.071 | - |
(S)-[2-[di(propan-2-yl)amino]ethyl] O,O-diethyl phosphorothioate | pH 9.0, 30°C, recombinant mutant I106G/F132V/H254Q/H257F/S308L | Brevundimonas diminuta | |
14.571 | - |
(S)-[2-[di(propan-2-yl)amino]ethyl] O,O-diethyl phosphorothioate | pH 9.0, 30°C, recombinant mutant A80V/F132V/K185R/H254Q/H257Y/I274N | Brevundimonas diminuta | |
17.1 | - |
4-acetylphenyl (S)-2-methylpropyl methylphosphonate | pH 9.0, 30°C, recombinant mutant A80V/F132V/K185R/H254Q/H257Y/I274N/S308L | Brevundimonas diminuta | |
19 | - |
(S)-[2-[di(propan-2-yl)amino]ethyl] O,O-diethyl phosphorothioate | pH 9.0, 30°C, recombinant mutant F132V/H254Q/H257F | Brevundimonas diminuta | |
21.053 | - |
(S)-[2-[di(propan-2-yl)amino]ethyl] O,O-diethyl phosphorothioate | pH 9.0, 30°C, recombinant mutant I106C/F132V/H254Q/H257F/S308L | Brevundimonas diminuta | |
21.4 | - |
4-acetylphenyl (R)-2-methylpropyl methylphosphonate | pH 9.0, 30°C, recombinant mutant A80V/I106C/F132V/K185R/H254Q/H257Y/I274N | Brevundimonas diminuta | |
21.539 | - |
(S)-[2-[di(propan-2-yl)amino]ethyl] O,O-diethyl phosphorothioate | pH 9.0, 30°C, recombinant mutant F132V/H254Q/H257F/S308L | Brevundimonas diminuta | |
25 | - |
4-acetylphenyl (R)-2-methylpropyl methylphosphonate | pH 9.0, 30°C, recombinant mutant I106G/F132V/H254Q/H257F | Brevundimonas diminuta | |
28.75 | - |
(S)-[2-[di(propan-2-yl)amino]ethyl] O,O-diethyl phosphorothioate | pH 9.0, 30°C, recombinant mutant A80V/I106C/F132V/K185R/H254Q/H257Y/I274N/S308L | Brevundimonas diminuta | |
30 | - |
(S)-[2-[di(propan-2-yl)amino]ethyl] O,O-diethyl phosphorothioate | pH 9.0, 30°C, recombinant mutant I106C/F132V/H254Q/H257Y/A270V/L272M/I274N/S308L/Y309F | Brevundimonas diminuta | |
30.137 | - |
(S)-[2-[di(propan-2-yl)amino]ethyl] O,O-diethyl phosphorothioate | pH 9.0, 30°C, recombinant mutant A80V/F132V/K185R/H254Q/H257Y/I274N/S308L | Brevundimonas diminuta | |
32.7 | - |
4-acetylphenyl (R)-2-methylpropyl methylphosphonate | pH 9.0, 30°C, recombinant mutant A80V/I106G/F132V/K185R/H254Q/H257Y/I274N/S308L | Brevundimonas diminuta | |
34 | - |
4-acetylphenyl (S)-2-methylpropyl methylphosphonate | pH 9.0, 30°C, recombinant mutant A80V/F132V/K185R/H254Q/H257Y/I274N | Brevundimonas diminuta | |
34.375 | - |
4-acetylphenyl (R)-2-methylpropyl methylphosphonate | pH 9.0, 30°C, recombinant mutant A80V/F132V/K185R/H254Q/H257Y/I274N/S308L | Brevundimonas diminuta | |
38.5 | - |
4-acetylphenyl (S)-2-methylpropyl methylphosphonate | pH 9.0, 30°C, recombinant mutant A80V/I106C/F132V/K185R/H254Q/H257Y/I274N | Brevundimonas diminuta | |
42.6 | - |
4-acetylphenyl (S)-2-methylpropyl methylphosphonate | pH 9.0, 30°C, recombinant mutant I106C/F132V/H254Q/H257F | Brevundimonas diminuta | |
49.4 | - |
4-acetylphenyl (R)-2-methylpropyl methylphosphonate | pH 9.0, 30°C, recombinant wild-type enzyme | Brevundimonas diminuta | |
61 | - |
4-acetylphenyl (R)-2-methylpropyl methylphosphonate | pH 9.0, 30°C, recombinant mutant I106C/F132V/H254Q/H257F | Brevundimonas diminuta | |
81.7 | - |
4-acetylphenyl (R)-2-methylpropyl methylphosphonate | pH 9.0, 30°C, recombinant mutant I106G/F132V/H254Q/H257Y/A270V/L272M/I274N/S308L | Brevundimonas diminuta | |
85 | - |
(S)-[2-[di(propan-2-yl)amino]ethyl] O,O-diethyl phosphorothioate | pH 9.0, 30°C, recombinant mutant I106C/F132V/H254Q/H257Y/A270V/L272M/I274N/S308L | Brevundimonas diminuta | |
94.1 | - |
4-acetylphenyl (R)-2-methylpropyl methylphosphonate | pH 9.0, 30°C, recombinant mutant A80V/F132V/K185R/H254Q/H257Y/I274N | Brevundimonas diminuta | |
110.5 | - |
4-acetylphenyl (S)-2-methylpropyl methylphosphonate | pH 9.0, 30°C, recombinant mutant I106C/F132V/H254Q/H257F/S308L | Brevundimonas diminuta | |
112 | - |
4-acetylphenyl (S)-2-methylpropyl methylphosphonate | pH 9.0, 30°C, recombinant mutant I106C/F132V/H254Q/H257Y/A270V/L272M/I274N/S308L | Brevundimonas diminuta | |
124.3 | - |
4-acetylphenyl (R)-2-methylpropyl methylphosphonate | pH 9.0, 30°C, recombinant mutant I106G/F132V/H254Q/H257F/S308L | Brevundimonas diminuta | |
143.5 | - |
4-acetylphenyl (S)-2-methylpropyl methylphosphonate | pH 9.0, 30°C, recombinant mutant A80V/I106C/F132V/K185R/H254Q/H257Y/I274N/S308L | Brevundimonas diminuta | |
158.3 | - |
4-acetylphenyl (R)-2-methylpropyl methylphosphonate | pH 9.0, 30°C, recombinant mutant H254Q/H257F | Brevundimonas diminuta | |
162.9 | - |
4-acetylphenyl (R)-2-methylpropyl methylphosphonate | pH 9.0, 30°C, recombinant mutant I106C/F132V/H254Q/H257Y/A270V/L272M/I274N/S308L | Brevundimonas diminuta | |
166.67 | - |
4-acetylphenyl (S)-2-methylpropyl methylphosphonate | pH 9.0, 30°C, recombinant mutant I106G/F132V/H254Q/H257F | Brevundimonas diminuta | |
180 | - |
4-acetylphenyl (S)-2-methylpropyl methylphosphonate | pH 9.0, 30°C, recombinant mutant I106G/F132V/H254Q/H257F/S308L | Brevundimonas diminuta | |
255.6 | - |
4-acetylphenyl (R)-2-methylpropyl methylphosphonate | pH 9.0, 30°C, recombinant mutant I106C/F132V/H254Q/H257F/S308L | Brevundimonas diminuta | |
290 | - |
4-acetylphenyl (S)-2-methylpropyl methylphosphonate | pH 9.0, 30°C, recombinant mutant H254Q/H257F | Brevundimonas diminuta | |
294.4 | - |
4-acetylphenyl (S)-2-methylpropyl methylphosphonate | pH 9.0, 30°C, recombinant mutant A80V/I106C/F132V/K185R/H254Q/H257Y/I274N | Brevundimonas diminuta | |
313 | - |
4-acetylphenyl (R)-2-methylpropyl methylphosphonate | pH 9.0, 30°C, recombinant mutant A80V/I106C/F132V/K185R/H254Q/H257Y/I274N/S308L | Brevundimonas diminuta | |
333.3 | - |
4-acetylphenyl (R)-2-methylpropyl methylphosphonate | pH 9.0, 30°C, recombinant mutant F132V/H254Q/H257F/S308L | Brevundimonas diminuta | |
535.5 | - |
4-acetylphenyl (S)-2-methylpropyl methylphosphonate | pH 9.0, 30°C, recombinant mutant I106G/F132V/H254Q/H257Y/A270V/L272M/I274N/S308L | Brevundimonas diminuta | |
883.3 | - |
4-acetylphenyl (S)-2-methylpropyl methylphosphonate | pH 9.0, 30°C, recombinant mutant A80V/I106G/F132V/K185R/H254Q/H257Y/I274N/S308L | Brevundimonas diminuta | |
1920 | - |
diethyl-paraoxon | pH 9.0, 30°C, recombinant mutant I106G/F132V/H254Q/H257F | Brevundimonas diminuta | |
4925 | - |
diethyl-paraoxon | pH 9.0, 30°C, recombinant mutant I106C/F132V/H254Q/H257F | Brevundimonas diminuta | |
6304 | - |
diethyl-paraoxon | pH 9.0, 30°C, recombinant mutant A80V/I106G/F132V/K185R/H254Q/H257Y/I274N | Brevundimonas diminuta | |
6786 | - |
diethyl-paraoxon | pH 9.0, 30°C, recombinant mutant I106C/F132V/H254Q/H257F/S308L | Brevundimonas diminuta | |
7736 | - |
diethyl-paraoxon | pH 9.0, 30°C, recombinant mutant H254Q/H257F | Brevundimonas diminuta | |
8824 | - |
diethyl-paraoxon | pH 9.0, 30°C, recombinant mutant A80V/F132V/K185R/H254Q/H257Y/I274N | Brevundimonas diminuta | |
9722 | - |
diethyl-paraoxon | pH 9.0, 30°C, recombinant mutant A80V/I106C/F132V/K185R/H254Q/H257Y/I274N | Brevundimonas diminuta | |
9818 | - |
diethyl-paraoxon | pH 9.0, 30°C, recombinant mutant F132V/H254Q/H257F | Brevundimonas diminuta | |
11806 | - |
diethyl-paraoxon | pH 9.0, 30°C, recombinant mutant I106C/F132V/H254Q/H257Y/A270V/L272M/I274N/S308L | Brevundimonas diminuta | |
12222 | - |
diethyl-paraoxon | pH 9.0, 30°C, recombinant mutant F132V/H254Q/H257F/S308L | Brevundimonas diminuta | |
14239 | - |
diethyl-paraoxon | pH 9.0, 30°C, recombinant mutant I106C/F132V/H254Q/H257Y/A270V/L272M/I274N/S308L/Y309F | Brevundimonas diminuta | |
14500 | - |
diethyl-paraoxon | pH 9.0, 30°C, recombinant mutant A80V/F132V/K185R/H254Q/H257Y/I274N/S308L | Brevundimonas diminuta | |
16515 | - |
diethyl-paraoxon | pH 9.0, 30°C, recombinant mutant I106G/F132V/H254Q/H257Y/A270V/L272M/I274N/S308L | Brevundimonas diminuta | |
19434 | - |
diethyl-paraoxon | pH 9.0, 30°C, recombinant mutant A80V/I106C/F132V/K185R/H254Q/H257Y/I274N/S308L | Brevundimonas diminuta | |
20727 | - |
diethyl-paraoxon | pH 9.0, 30°C, recombinant mutant I106G/F132V/H254Q/H257F/S308L | Brevundimonas diminuta | |
21239 | - |
diethyl-paraoxon | pH 9.0, 30°C, recombinant mutant A80V/I106G/F132V/K185R/H254Q/H257Y/I274N/S308L | Brevundimonas diminuta | |
27531 | - |
diethyl-paraoxon | pH 9.0, 30°C, recombinant wild-type enzyme | Brevundimonas diminuta |