Ene-reductase from Geobacillus sp. 30

Enzyme Description

Species
Geobacillus sp. 30 β†—
Extremophile
No but "thermotolerant" enzyme
EC Number
1.6.99.1 β†—

Sequence

Length: 340 amino acids
METMLFSPYTIRGVTLKNRIVMSPMCMYSCDTEDGKVRTWHKIHYPTRAVGQVGLIIVEATAVTAQGRISARDLGIWSDEHVDGLRELVSLVKEHGAKIGIQLAHAGRKAEVEGEIIAPSAIPFNEKTRTPKEMTKTDIEETIQAFQNGARRAKEAGFDVIEIHGAHGYLINEFLSPLANKRQDEYGGSSENRYRFLGEVIDAVRQVWDGPLFVRISASDYHPEGLTAKDYIPYAKRMKEQGVDLIDVSSGAVVPATINAYPGYQVPFAELIRREAEIATGAVGLITSGWQAEEILRNGRADLVFLARELLRNPYWPYAAAKELGTTIPAPVQYERGWRF
Naoto Tsuji et al. (2014) β€” Isolation and characterization of a thermotolerant ene reductase from Geobacillus sp. 30 and its heterologous expression in Rhodococcus opacus
Applied Microbiology and Biotechnology  Β· doi:10.1007/s00253-014-5668-9 β†—  Β· Stability - Incubation
16 measurements ▼
Database ID
UniProt: W0T2Q3 β†—
Sequence Annotation
Explicit - Provided GenBank Accession Number
Protein Source
Recombinant, host bacterium Rhodococcus opacus B-4

Experimental Data (16 measurements)

16 measurements
Property Assay Solvent Solvent Volume Aqueous Reference Measured Value Units Solution pH Temperature Substrate(s) Product(s) Cofactor(s) Shaking Comments
Stability - Incubation Activity after incubation (12 hours at 30Β°C), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in aqueous phase Methanol 5% 100 88 % Incubation: ?, Assay: 20 mM bicine-NaOH Incubation: ?, Assay: 8 Incubation: 30Β°C, Assay: 50Β°C 2 mM Ketoisophorone Levodione 0.2 mM NADPH Incubation: 1400 rpm Non-incubated control (in %), assay measured in aqueous phase | Uncertainty about whether assay was in the presence of organic solvent or in aqueous phase
Stability - Incubation Activity after incubation (12 hours at 30Β°C), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in aqueous phase Methanol 10% 100 86 % Incubation: ?, Assay: 20 mM bicine-NaOH Incubation: ?, Assay: 8 Incubation: 30Β°C, Assay: 50Β°C 2 mM Ketoisophorone Levodione 0.2 mM NADPH Incubation: 1400 rpm Non-incubated control (in %), assay measured in aqueous phase | Uncertainty about whether assay was in the presence of organic solvent or in aqueous phase
Stability - Incubation Activity after incubation (12 hours at 30Β°C), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in aqueous phase Methanol 20% 100 83 % Incubation: ?, Assay: 20 mM bicine-NaOH Incubation: ?, Assay: 8 Incubation: 30Β°C, Assay: 50Β°C 2 mM Ketoisophorone Levodione 0.2 mM NADPH Incubation: 1400 rpm Non-incubated control (in %), assay measured in aqueous phase | Uncertainty about whether assay was in the presence of organic solvent or in aqueous phase
Stability - Incubation Activity after incubation (12 hours at 30Β°C), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in aqueous phase Dimethyl Sulfoxide (DMSO) 5% 100 90 % Incubation: ?, Assay: 20 mM bicine-NaOH Incubation: ?, Assay: 8 Incubation: 30Β°C, Assay: 50Β°C 2 mM Ketoisophorone Levodione 0.2 mM NADPH Incubation: 1400 rpm Non-incubated control (in %), assay measured in aqueous phase | Uncertainty about whether assay was in the presence of organic solvent or in aqueous phase
Stability - Incubation Activity after incubation (12 hours at 30Β°C), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in aqueous phase Dimethyl Sulfoxide (DMSO) 10% 100 89 % Incubation: ?, Assay: 20 mM bicine-NaOH Incubation: ?, Assay: 8 Incubation: 30Β°C, Assay: 50Β°C 2 mM Ketoisophorone Levodione 0.2 mM NADPH Incubation: 1400 rpm Non-incubated control (in %), assay measured in aqueous phase | Uncertainty about whether assay was in the presence of organic solvent or in aqueous phase
Stability - Incubation Activity after incubation (12 hours at 30Β°C), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in aqueous phase Dimethyl Sulfoxide (DMSO) 20% 100 69 % Incubation: ?, Assay: 20 mM bicine-NaOH Incubation: ?, Assay: 8 Incubation: 30Β°C, Assay: 50Β°C 2 mM Ketoisophorone Levodione 0.2 mM NADPH Incubation: 1400 rpm Non-incubated control (in %), assay measured in aqueous phase | Uncertainty about whether assay was in the presence of organic solvent or in aqueous phase
Stability - Incubation Activity after incubation (12 hours at 30Β°C), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in aqueous phase Cyclohexanol 5% 100 84 % Incubation: ?, Assay: 20 mM bicine-NaOH Incubation: ?, Assay: 8 Incubation: 30Β°C, Assay: 50Β°C 2 mM Ketoisophorone Levodione 0.2 mM NADPH Incubation: 1400 rpm Non-incubated control (in %), assay measured in aqueous phase | Uncertainty about whether assay was in the presence of organic solvent or in aqueous phase
Stability - Incubation Activity after incubation (12 hours at 30Β°C), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in aqueous phase Cyclohexanol 10% 100 72 % Incubation: ?, Assay: 20 mM bicine-NaOH Incubation: ?, Assay: 8 Incubation: 30Β°C, Assay: 50Β°C 2 mM Ketoisophorone Levodione 0.2 mM NADPH Incubation: 1400 rpm Non-incubated control (in %), assay measured in aqueous phase | Uncertainty about whether assay was in the presence of organic solvent or in aqueous phase
Stability - Incubation Activity after incubation (12 hours at 30Β°C), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in aqueous phase Cyclohexanol 20% 100 64 % Incubation: ?, Assay: 20 mM bicine-NaOH Incubation: ?, Assay: 8 Incubation: 30Β°C, Assay: 50Β°C 2 mM Ketoisophorone Levodione 0.2 mM NADPH Incubation: 1400 rpm Non-incubated control (in %), assay measured in aqueous phase | Uncertainty about whether assay was in the presence of organic solvent or in aqueous phase
Stability - Incubation Activity after incubation (12 hours at 30Β°C), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in aqueous phase Cyclohexane 5% 100 94 % Incubation: ?, Assay: 20 mM bicine-NaOH Incubation: ?, Assay: 8 Incubation: 30Β°C, Assay: 50Β°C 2 mM Ketoisophorone Levodione 0.2 mM NADPH Incubation: 1400 rpm Non-incubated control (in %), assay measured in aqueous phase | Uncertainty about whether assay was in the presence of organic solvent or in aqueous phase
Stability - Incubation Activity after incubation (12 hours at 30Β°C), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in aqueous phase Cyclohexane 10% 100 90 % Incubation: ?, Assay: 20 mM bicine-NaOH Incubation: ?, Assay: 8 Incubation: 30Β°C, Assay: 50Β°C 2 mM Ketoisophorone Levodione 0.2 mM NADPH Incubation: 1400 rpm Non-incubated control (in %), assay measured in aqueous phase | Uncertainty about whether assay was in the presence of organic solvent or in aqueous phase
Stability - Incubation Activity after incubation (12 hours at 30Β°C), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in aqueous phase Cyclohexane 20% 100 89 % Incubation: ?, Assay: 20 mM bicine-NaOH Incubation: ?, Assay: 8 Incubation: 30Β°C, Assay: 50Β°C 2 mM Ketoisophorone Levodione 0.2 mM NADPH Incubation: 1400 rpm Non-incubated control (in %), assay measured in aqueous phase | Uncertainty about whether assay was in the presence of organic solvent or in aqueous phase
Stability - Incubation Activity after incubation (6 hours at 50Β°C), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in aqueous phase Cyclohexanol 50% 100 86 % Incubation: ?, Assay: 20 mM bicine-NaOH Incubation: ?, Assay: 8 Incubation: 50Β°C, Assay: 50Β°C 2 mM Ketoisophorone Levodione 0.2 mM NADPH Incubation: 1400 rpm Non-incubated control (in %), assay measured in aqueous phase | Uncertainty about whether assay was in the presence of organic solvent or in aqueous phase
Stability - Incubation Activity after incubation (12 hours at 50Β°C), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in aqueous phase Cyclohexanol 50% 103 73 % Incubation: ?, Assay: 20 mM bicine-NaOH Incubation: ?, Assay: 8 Incubation: 50Β°C, Assay: 50Β°C 2 mM Ketoisophorone Levodione 0.2 mM NADPH Incubation: 1400 rpm Non-incubated control (in %), assay measured in aqueous phase | Uncertainty about whether assay was in the presence of organic solvent or in aqueous phase
Stability - Incubation Activity after incubation (18 hours at 50Β°C), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in aqueous phase Cyclohexanol 50% 82 70 % Incubation: ?, Assay: 20 mM bicine-NaOH Incubation: ?, Assay: 8 Incubation: 50Β°C, Assay: 50Β°C 2 mM Ketoisophorone Levodione 0.2 mM NADPH Incubation: 1400 rpm Non-incubated control (in %), assay measured in aqueous phase | Uncertainty about whether assay was in the presence of organic solvent or in aqueous phase
Stability - Incubation Activity after incubation (24 hours at 50Β°C), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in aqueous phase Cyclohexanol 50% 69 50 % Incubation: ?, Assay: 20 mM bicine-NaOH Incubation: ?, Assay: 8 Incubation: 50Β°C, Assay: 50Β°C 2 mM Ketoisophorone Levodione 0.2 mM NADPH Incubation: 1400 rpm Non-incubated control (in %), assay measured in aqueous phase | Uncertainty about whether assay was in the presence of organic solvent or in aqueous phase

Visualization : Stability β€” Incubation

One bar per measurement. Colour = solvent, shade = solvent volume.

Structure

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