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 (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 | 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 | 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 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 | UniProt | Comment | Textmining |
---|---|---|---|
Brevundimonas diminuta | - |
- |
- |
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 | 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 | 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 | Comment | Organism |
---|---|---|
OPH | - |
Brevundimonas diminuta |
organophosphate hydrolase | - |
Brevundimonas diminuta |
phosphotriesterase | - |
Brevundimonas diminuta |
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 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 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 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 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 |
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 |