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Literature summary for 3.1.8.1 extracted from
- Insertion of an unnatural amino acid into the protein structure: preparation and properties of 3-fluorotyrosine-containing organophosphate hydrolase (2006), Russ. Chem. Bull., 55, 369-374.
No PubMed abstract available
Activating Compound
| Activating Compound | Comment | Organism | Structure |
|---|---|---|---|
| additional information | efficient enzyme-catalyzed hydrolysis of OPC requires essential stabilization of the enzyme in the acidic pH region | Brevundimonas diminuta |
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| expression of N-terminally His6-tagged wild-type and 3-fluorotyrosine-containing OPH in Escherichia coli strain B-2935 devoid of the metabolic ways of tyrosine biosynthesis, the presence of 3-F-Tyr in the medium inhibits the cell growth | Brevundimonas diminuta |
Protein Variants
| Protein Variants | Comment | Organism |
|---|---|---|
| additional information | construction of a 3-fluorotyrosine-containing enzyme by replacement of Tyr residues, Tyr292, Tyr239, and Tyr248 residues are completely replaced by the fluorinated analogue, while the Tyr156 residue is replaced by 90%, overview, the substitution extends substantially the pH range of action of the fluorine-substituted enzyme compared with the unsubstituted one, the mutant shows increased pH and temperature stability, but reduced activity | Brevundimonas diminuta |
General Stability
| General Stability | Organism |
|---|---|
| efficient enzyme-catalyzed hydrolysis of OPC requires essential stabilization of the enzyme in the acidic pH region | Brevundimonas diminuta |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| additional information | - |
additional information | kinetics of recombinant His6-tagged wild-type OPH and 3-fluorotyrosine-containing recombinant His6-tagged OPH | Brevundimonas diminuta | |
| 0.01 | - |
paraoxon | pH 10.5, 25°C, recombinant His6-tagged wild-type OPH | Brevundimonas diminuta |  |
| 0.104 | - |
paraoxon | pH 10.5, 25°C, 3-fluorotyrosine-containing recombinant His6-tagged OPH | Brevundimonas diminuta | |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Brevundimonas diminuta | - |
- |
- |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| recombinant N-terminally His6-tagged OPH from Escherichia coli strain B-2935 by Co2+-iminodiacetic acid-polyacrylamide cryogel affinity chromatography | Brevundimonas diminuta |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| additional information | OPH catalyzes hydrolysis of organophosphorus compounds, OPC15-20, to give phosphoric or alkylphosphonic acid derivatives, which is accompanied by a decrease in the pH of the reaction medium through release of two protons | Brevundimonas diminuta | ? | - |
? | |
| paraoxon + H2O | i.e. diethyl-4-nitrophenyl phosphate | Brevundimonas diminuta | diethylphosphate + 4-nitrophenol | - |
? | |
Subunits
| Subunits | Comment | Organism |
|---|---|---|
| ? | x * 37691.1, recombinant N-terminally His6-tagged mutant 3-fluorotyrosine-containing OPH, mass spectrometry, x * 37602.9, recombinant N-terminally His6-tagged wild-type OPH, mass spectrometry | Brevundimonas diminuta |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| OPH | - |
Brevundimonas diminuta |
| organophosphate hydrolase | - |
Brevundimonas diminuta |
| phosphotriesterase | - |
Brevundimonas diminuta |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 45 | 50 | 3-fluorotyrosine-containing recombinant His6-tagged OPH | Brevundimonas diminuta |
| 57 | 62 | recombinant His6-tagged wild-type OPH | Brevundimonas diminuta |
Temperature Stability [°C]
| Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 50 | - |
15 min, purified recombinant His6-tagged wild-type OPH, 50% remaining activity | Brevundimonas diminuta |
| 60 | - |
15 min, purified 3-fluorotyrosine-containing recombinant His6-tagged OPH, stable up to | Brevundimonas diminuta |
| 75 | - |
15 min, purified 3-fluorotyrosine-containing recombinant His6-tagged OPH, loss of 20% activity | Brevundimonas diminuta |
| 85 | - |
15 min, purified 3-fluorotyrosine-containing recombinant His6-tagged OPH, complete inactivation | Brevundimonas diminuta |
Turnover Number [1/s]
| Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 125 | - |
paraoxon | pH 10.5, 25°C, 3-fluorotyrosine-containing recombinant His6-tagged OPH | Brevundimonas diminuta | |
| 5100 | - |
paraoxon | pH 10.5, 25°C, recombinant His6-tagged wild-type OPH | Brevundimonas diminuta | |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 7.5 | 10.5 | 3-fluorotyrosine-containing recombinant His6-tagged OPH | Brevundimonas diminuta |
| 9.5 | 10.5 | recombinant His6-tagged wild-type OPH | Brevundimonas diminuta |
pH Range
| pH Minimum | pH Maximum | Comment | Organism |
|---|---|---|---|
| 5.5 | 12 | the fluorinated enzyme exhibits stable catalytic activity in the pH range, 35-50% of maximal activity at pH 5.5 and pH 12.0 | Brevundimonas diminuta |
| 7 | 12 | wild-type enzyme | Brevundimonas diminuta |
pI Value
| Organism | Comment | pI Value Maximum | pI Value |
|---|---|---|---|
| Brevundimonas diminuta | the fluorinated enzyme exhibits stable catalytic activity in the pH range, 35-50% of maximal activity at pH 5.5 and pH 12.0 | - |
7.6 |
| Brevundimonas diminuta | wild-type enzyme | - |
8.5 |
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