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Literature summary for 3.1.8.1 extracted from
- Functional analysis of organophosphorus hydrolase variants with high degradation activity towards organophosphate pesticides (2006), Protein Eng. Des. Sel., 19, 99-105.
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| expression of His-tagged wild-type and mutants | Escherichia coli K-12 |
| gene opd, expression of His-tagged wild-type and mutant enzymes | Escherichia coli |
Protein Variants
| Protein Variants | Comment | Organism |
|---|---|---|
| 185X | construction of diverse mutants with polymorphism at residue 185 showing altered activities with substrate methyl parathion and paraoxon, overview | Escherichia coli K-12 |
| A14T/A80V/K185R/H257Y/I274N | OPH genetic variant 22A11, constructed by site directed mutagenesis, the variant shows altered kinetics compared to the wild-type enzyme | Escherichia coli K-12 |
| A14T/A80V/K185R/H257Y/I274N | random saturation mutagenesis, variant 2H2, increased activity and altered kinetics compared to the wild-type enzyme | Escherichia coli |
| A80V | OPH genetic variant 2H2, site directed mutagenesis, the variant shows altered kinetics compared to the wild-type enzyme | Escherichia coli K-12 |
| A80V/I274N/K185R | site-directed mutagenesis, increased activity and altered kinetics compared to the wild-type enzyme | Escherichia coli |
| A80V/S365P | random saturation mutagenesis, variant 2H2, increased activity and altered kinetics compared to the wild-type enzyme | Escherichia coli |
| H257Y | OPH genetic variant 6D4, site directed mutagenesis, the variant shows altered kinetics compared to the wild-type enzyme | Escherichia coli K-12 |
| H257Y | random saturation mutagenesis, variant 6D4, increased activity and altered kinetics compared to the wild-type enzyme | Escherichia coli |
| I274N | OPH genetic variant 5A6, site directed mutagenesis, the variant shows altered kinetics compared to the wild-type enzyme | Escherichia coli K-12 |
| I274N | random saturation mutagenesis, variant 5A6, increased activity and altered kinetics compared to the wild-type enzyme | Escherichia coli |
| I274N/H257Y | OPH genetic variant 21E1, site directed mutagenesis, the variant shows altered kinetics compared to the wild-type enzyme | Escherichia coli K-12 |
| I274N/K185R | site-directed mutagenesis, increased activity and altered kinetics compared to the wild-type enzyme | Escherichia coli |
| K185E | site-directed mutagenesis, increased activity and altered kinetics compared to the wild-type enzyme | Escherichia coli |
| K185R | site-directed mutagenesis, increased activity and altered kinetics compared to the wild-type enzyme | Escherichia coli |
| L182S/V310A | random saturation mutagenesis, variant 2F6, increased activity and altered kinetics compared to the wild-type enzyme | Escherichia coli |
| additional information | construction of genetic variants of OPH wild-type enzyme with altered sbstrate specificities, overview | Escherichia coli K-12 |
| additional information | construction of diverse mutant variants by saturation mutagenesis, e.g. mutant 22A11, sequencing of randomly chosen mutants, the mutants show increased activity and altered kinetics compared to the wild-type enzyme, overview | Escherichia coli |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| additional information | - |
additional information | kinetics for diverse random mutant variants, overview | Escherichia coli | |
| 0.12 | - |
paraoxon | pH 8.0, 37°C, recombinant His6-tagged wild-type OPH | Escherichia coli |  |
| 1.22 | - |
methylparathion | pH 8.0, 37°C, recombinant His6-tagged wild-type OPH | Escherichia coli | |
Metals/Ions
| Metals/Ions | Comment | Organism | Structure |
|---|---|---|---|
| Co2+ | activates | Escherichia coli | |
| Co2+ | activates | Escherichia coli K-12 | |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Escherichia coli | - |
gene opd | - |
| Escherichia coli K-12 | - |
gene opd, wild-type and variant 22A11 | - |
| Escherichia coli K-12 XL1-Blue | - |
gene opd, wild-type and variant 22A11 | - |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| recombinant His-tagged wild-type and mutant enzymes by nickel affinity chromatography | Escherichia coli |
Specific Activity [micromol/min/mg]
| Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
|---|---|---|---|
| additional information | - |
- |
Escherichia coli |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| chlorpyrifos + H2O | the wild-type enzyme shows lower activity compared to the activity of genetic variant 22A11, overview | Escherichia coli K-12 | 3,5,6-trichloro-pyridin-2-ol + diethyl thiophosphate | - |
? | |
| chlorpyrifos + H2O | the wild-type enzyme shows lower activity compared to the activity of genetic variant 22A11, overview | Escherichia coli K-12 XL1-Blue | 3,5,6-trichloro-pyridin-2-ol + diethyl thiophosphate | - |
? | |
| coumaphos + H2O | - |
Escherichia coli | ? | - |
? | |
| coumaphos + H2O | - |
Escherichia coli K-12 | 3-chloro-4-methylumbelliferone + diethyl thiophosphate | - |
? | |
| methyl parathion + H2O | - |
Escherichia coli K-12 | 4-nitrophenol + dimethyl thiophosphate | - |
? | |
| methyl parathion + H2O | - |
Escherichia coli K-12 XL1-Blue | 4-nitrophenol + dimethyl thiophosphate | - |
? | |
| methyl parathion + H2O | - |
Escherichia coli | dimethyl thiophosphate + 4-nitrophenol | - |
? | |
| additional information | substrate specificities of wild-type enzyme and genetic variants, overview | Escherichia coli K-12 | ? | - |
? | |
| additional information | substrate specificities of wild-type enzyme and genetic variants, overview | Escherichia coli K-12 XL1-Blue | ? | - |
? | |
| paraoxon + H2O | - |
Escherichia coli | diethylphosphate + 4-nitrophenol | - |
? | |
| paraoxon + H2O | - |
Escherichia coli K-12 | 4-nitrophenol + diethyl phosphate | - |
? | |
| paraoxon + H2O | - |
Escherichia coli K-12 XL1-Blue | 4-nitrophenol + diethyl phosphate | - |
? | |
| parathion + H2O | - |
Escherichia coli K-12 | 4-nitrophenol + diethyl thiophosphate | - |
? | |
| parathion + H2O | - |
Escherichia coli K-12 XL1-Blue | 4-nitrophenol + diethyl thiophosphate | - |
? | |
| parathion + H2O | - |
Escherichia coli | diethylthiophosphate + 4-nitrophenol | - |
? | |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| OPH | - |
Escherichia coli |
| OPH | - |
Escherichia coli K-12 |
| organophosphorus hydrolase | - |
Escherichia coli |
| organophosphorus hydrolase | - |
Escherichia coli K-12 |
| phosphotriesterase | - |
Escherichia coli |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 37 | - |
assay at | Escherichia coli |
| 37 | - |
assay at | Escherichia coli K-12 |
Turnover Number [1/s]
| Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 2300 | - |
methylparathion | pH 8.0, 37°C, recombinant His6-tagged wild-type OPH | Escherichia coli | |
| 11700 | - |
paraoxon | pH 8.0, 37°C, recombinant His6-tagged wild-type OPH | Escherichia coli | |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 8 | - |
- |
Escherichia coli K-12 |
| 8 | - |
assay at | Escherichia coli |
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