Ene-reductase (gene oyeRo2a) from Rhodococcus opacus 1CP

Enzyme Description

Species
Rhodococcus opacus 1CP β†—
Extremophile
No
EC Number
1.6.99.1 β†—

Sequence

Length: 366 amino acids
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
21 measurements ▼
Database ID
UniParc: UPI0007209AF6 β†—
Sequence Annotation
Explicit - Provided
Protein Source
Recombinant, host bacterium Escherichia coli BL21 (DE3)

Experimental Data (21 measurements)

21 measurements β€” page 1 of 2
Property Assay Solvent Solvent Volume Aqueous Reference Measured Value Units Extraction method Data type 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 25 % v/v 100 27 % Digital Continuous 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 % v/v 100 0 % Digital Continuous 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 % v/v 100 50 % Digital Continuous 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 % v/v 100 99 % Digital Continuous 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 30 % v/v 100 7 % Digital Continuous 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 % v/v 100 106 % Digital Continuous 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 % v/v 100 54 % Digital Continuous 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 % v/v 100 83 % Digital Continuous 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 % v/v 100 100 % Digital Continuous 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 % v/v 100 101 % Digital Continuous 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 % v/v 100 12 % Digital Continuous 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 % v/v 100 101 % Digital Continuous 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 % v/v 100 36 % Digital Continuous 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 % v/v 100 85 % Digital Continuous 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 % v/v 100 85 % Digital Continuous 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 % v/v 100 97 % Digital Continuous 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 % v/v 100 86 % Digital Continuous 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 % v/v 100 61 % Digital Continuous 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 % v/v 100 15 % Digital Continuous 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 (15 minutes at 25Β°C), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in aqueous phase Ethanol 20 % v/v 100 6 % Digital Continuous 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
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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.

Structure

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