Ene-reductase (gene oyeRo2a) from Rhodococcus opacus 1CP
Enzyme Description
Sequence
MSVLFEPITFRGVTVPNRVWMAPMCQYSADVTGRDVGVPGDWHRTHLVTRAIGGAGLILTEATAVSPEGRISPADLGIWNDTQTEAFAEINRRIREFGAVPGIQLGHAGRKGSAHVPWRGGGSLDGDDRLSWQTVAPSAIGFGDHTPPAAATTADIRKVVADFAAAAERASRAGFKVVEIHAAHGYLLHQFLSPVSNHRTDEYGGSFAGRIRLLLEVVDAVRGVWPAELPVFVRVSATDWLSEEPGLDADSWTPDQTVSLVQALADLGVDLVDVSSGGVASARIPIGPGYQVPFARRIQNETTVPAAAVGLITEPEQAERIVESGEAVAVFLGRELLRDPYWPRKAALVLNAQVTPQIPAQYARAY
Anika Riedel et al. (2015)
β Functional characterization and stability improvement of a 'thermophilic-like ' ene-reductase from Rhodococcus opacus 1CP
Frontiers in Microbiology
Β· doi:10.3389/fmicb.2015.01073 β
Β· Activity - Classical Stability - Incubation
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Database ID
UniParc: UPI0007209AF6 β
Sequence Annotation
Explicit - Provided
Protein Source
Recombinant, host bacterium Escherichia coli BL21 (DE3)
Experimental Data (22 measurements)
22 measurements β page 1 of 2
| Property | Assay | Solvent | Solvent Volume | Aqueous Reference | Measured Value | Units | Solution | pH | Temperature | Substrate(s) | Product(s) | Cofactor(s) | Shaking | Comments |
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| Activity - Classical | Activity measured by absorbance spectrophotometry (NADPH absorance measurement, 340 nm) in the presence of organic solvent | Ethanol | 30% | 100 | 7 | % | 25 mM phosphate buffer | 7.1 | 25Β°C | 1 mM Maleimide | Succinimide | 140 ΞΌM NADPH | β | Classical aqueous control (in %) |
| Activity - Classical | Activity measured by absorbance spectrophotometry (NADPH absorance measurement, 340 nm) in the presence of organic solvent | Dimethyl Sulfoxide (DMSO) | 40% | 100 | 0 | % | 25 mM phosphate buffer | 7.1 | 25Β°C | 1 mM Maleimide | Succinimide | 140 ΞΌM NADPH | β | Classical aqueous control (in %) |
| Activity - Classical | Activity measured by absorbance spectrophotometry (NADPH absorance measurement, 340 nm) in the presence of organic solvent | Dimethyl Sulfoxide (DMSO) | 30% | 100 | 50 | % | 25 mM phosphate buffer | 7.1 | 25Β°C | 1 mM Maleimide | Succinimide | 140 ΞΌM NADPH | β | Classical aqueous control (in %) |
| Activity - Classical | Activity measured by absorbance spectrophotometry (NADPH absorance measurement, 340 nm) in the presence of organic solvent | Dimethyl Sulfoxide (DMSO) | 20% | 100 | 99 | % | 25 mM phosphate buffer | 7.1 | 25Β°C | 1 mM Maleimide | Succinimide | 140 ΞΌM NADPH | β | Classical aqueous control (in %) |
| Activity - Classical | Activity measured by absorbance spectrophotometry (NADPH absorance measurement, 340 nm) in the presence of organic solvent | Dimethyl Sulfoxide (DMSO) | 10% | 100 | 101 | % | 25 mM phosphate buffer | 7.1 | 25Β°C | 1 mM Maleimide | Succinimide | 140 ΞΌM NADPH | β | Classical aqueous control (in %) |
| Activity - Classical | Activity measured by absorbance spectrophotometry (NADPH absorance measurement, 340 nm) in the presence of organic solvent | Ethanol | 10% | 100 | 106 | % | 25 mM phosphate buffer | 7.1 | 25Β°C | 1 mM Maleimide | Succinimide | 140 ΞΌM NADPH | β | Classical aqueous control (in %) |
| Activity - Classical | Activity measured by absorbance spectrophotometry (NADPH absorance measurement, 340 nm) in the presence of organic solvent | Ethanol | 25% | 100 | 27 | % | 25 mM phosphate buffer | 7.1 | 25Β°C | 1 mM Maleimide | Succinimide | 140 ΞΌM NADPH | β | Classical aqueous control (in %) |
| Activity - Classical | Activity measured by absorbance spectrophotometry (NADPH absorance measurement, 340 nm) in the presence of organic solvent | Ethanol | 20% | 100 | 54 | % | 25 mM phosphate buffer | 7.1 | 25Β°C | 1 mM Maleimide | Succinimide | 140 ΞΌM NADPH | β | Classical aqueous control (in %) |
| Activity - Classical | Activity measured by absorbance spectrophotometry (NADPH absorance measurement, 340 nm) in the presence of organic solvent | Ethanol | 15% | 100 | 83 | % | 25 mM phosphate buffer | 7.1 | 25Β°C | 1 mM Maleimide | Succinimide | 140 ΞΌM NADPH | β | Classical aqueous control (in %) |
| Activity - Classical | Activity measured by absorbance spectrophotometry (NADPH absorance measurement, 340 nm) in the presence of organic solvent | Ethanol | 10% | 100 | 100 | % | 25 mM phosphate buffer | 7.1 | 25Β°C | 1 mM Maleimide | Succinimide | 140 ΞΌM NADPH | β | Classical aqueous control (in %) |
| Activity - Classical | Activity measured by absorbance spectrophotometry (NADPH absorance measurement, 340 nm) in the presence of organic solvent | Ethanol | 5% | 100 | 101 | % | 25 mM phosphate buffer | 7.1 | 25Β°C | 1 mM Maleimide | Succinimide | 140 ΞΌM NADPH | β | Classical aqueous control (in %) |
| Activity - Classical | Activity measured by absorbance spectrophotometry (NADPH absorance measurement, 340 nm) in the presence of organic solvent | Dimethylformamide (DMF) | 10% | 100 | 12 | % | 25 mM phosphate buffer | 7.1 | 25Β°C | 1 mM Maleimide | Succinimide | 140 ΞΌM NADPH | β | Classical aqueous control (in %) |
| Activity - Classical | Activity measured by absorbance spectrophotometry (NADPH absorance measurement, 340 nm) in the presence of organic solvent | Dimethyl Sulfoxide (DMSO) | 10% | 100 | 101 | % | 25 mM phosphate buffer | 7.1 | 25Β°C | 1 mM Maleimide | Succinimide | 140 ΞΌM NADPH | β | Classical aqueous control (in %) |
| Activity - Classical | Activity measured by absorbance spectrophotometry (NADPH absorance measurement, 340 nm) in the presence of organic solvent | Acetonitrile | 10% | 100 | 36 | % | 25 mM phosphate buffer | 7.1 | 25Β°C | 1 mM Maleimide | Succinimide | 140 ΞΌM NADPH | β | Classical aqueous control (in %) |
| Activity - Classical | Activity measured by absorbance spectrophotometry (NADPH absorance measurement, 340 nm) in the presence of organic solvent | Acetone | 10% | 100 | 85 | % | 25 mM phosphate buffer | 7.1 | 25Β°C | 1 mM Maleimide | Succinimide | 140 ΞΌM NADPH | β | Classical aqueous control (in %) |
| Activity - Classical | Activity measured by absorbance spectrophotometry (NADPH absorance measurement, 340 nm) in the presence of organic solvent | Methanol | 10% | 100 | 85 | % | 25 mM phosphate buffer | 7.1 | 25Β°C | 1 mM Maleimide | Succinimide | 140 ΞΌM NADPH | β | Classical aqueous control (in %) |
| Stability - Incubation | Activity after incubation (1 minute at 25Β°C), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in aqueous phase | Ethanol | 20% | 100 | 97 | % | Incubation: 25 mM phosphate buffer, Assay: 25 mM phosphate buffer | Incubation: 7.1, Assay: 7.1 | Incubation: 25Β°C, Assay: 25Β°C | 1 mM Maleimide | Succinimide | Assay: 140 ΞΌM NADPH | β | Non-incubated control (in %), assay in aqueous phase | Uncertainty about whether the assay was in aqueous phase or in the presence of organic solvent |
| Stability - Incubation | Activity after incubation (3 minutes at 25Β°C), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in aqueous phase | Ethanol | 20% | 100 | 86 | % | Incubation: 25 mM phosphate buffer, Assay: 25 mM phosphate buffer | Incubation: 7.1, Assay: 7.1 | Incubation: 25Β°C, Assay: 25Β°C | 1 mM Maleimide | Succinimide | Assay: 140 ΞΌM NADPH | β | Non-incubated control (in %), assay in aqueous phase | Uncertainty about whether the assay was in aqueous phase or in the presence of organic solvent |
| Stability - Incubation | Activity after incubation (5 minutes at 25Β°C), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in aqueous phase | Ethanol | 20% | 100 | 61 | % | Incubation: 25 mM phosphate buffer, Assay: 25 mM phosphate buffer | Incubation: 7.1, Assay: 7.1 | Incubation: 25Β°C, Assay: 25Β°C | 1 mM Maleimide | Succinimide | Assay: 140 ΞΌM NADPH | β | Non-incubated control (in %), assay in aqueous phase | Uncertainty about whether the assay was in aqueous phase or in the presence of organic solvent |
| Stability - Incubation | Activity after incubation (10 minutes at 25Β°C), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in aqueous phase | Ethanol | 20% | 100 | 15 | % | Incubation: 25 mM phosphate buffer, Assay: 25 mM phosphate buffer | Incubation: 7.1, Assay: 7.1 | Incubation: 25Β°C, Assay: 25Β°C | 1 mM Maleimide | Succinimide | Assay: 140 ΞΌM NADPH | β | Non-incubated control (in %), assay in aqueous phase | Uncertainty about whether the assay was in aqueous phase or in the presence of organic solvent |
Visualization : Activity β Classical
One bar per measurement. Colour = solvent, shade = solvent volume.
Visualization : Stability β Incubation
One bar per measurement. Colour = solvent, shade = solvent volume.