Short-chain dehydrogenase/reductase (gene Is2-SDR) from a hot spring metagenomic library

Enzyme Description

Species
Na (hot spring metagenomic library)
Extremophile
Yes organism from a hot spring, thermophilic enzyme
EC Number
1.1.1.184 β†—

Sequence

Length: 253 amino acids
MKAVKLLDGKIALITGASKGIGRAIALGMADVGADIIATARTELELRELCDEIRMKGRRAEYIVADLRSTEEAKEMARKALGIFGRVDILVNNAGVSYPQPALETTEDRWDETFQINLKSLFFLTQIIARAMVEQGGGKIINISSQAGLVGLEDHAAYCASKGALILLTKVLAIEWGKYKINVNAVAPTIIMTPMAERAWADAKKREEALKKIPLGRFGQPEDVVGAVVFLASPYADLITGAVIPIDGGYTAQ
Erica Elisa Ferrandi et al. (2022) β€” Functional Characterization and Synthetic Application of Is2-SDR, a Novel Thermostable and Promiscuous Ketoreductase from a Hot Spring Metagenome
International Journal of Molecular Sciences  Β· doi:10.3390/ijms232012153 β†—  Β· Stability - Incubation
68 measurements ▼
Database ID
UniParc: UPI001862B743 β†—
Sequence Annotation
Explicit - Provided
Protein Source
Recombinant, host bacterium Escherichia coli BL21 (DE3)

Experimental Data (68 measurements)

68 measurements β€” page 2 of 4
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 (3 hours at 25Β°C), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in aqueous phase Cyclopentyl Methyl Ether (CPME) 50% 86 94 % Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer Incubation: 7, Assay: 7 Incubation: 25Β°C, Assay: Room temperature 2 mM Ethyl 3-methyl-2-oxobutyrate Ethyl 3-methyl-2-hydroxybutyrate 0.2 mM NADPH Incubation: 100 rpm Water-incubated control (in %), assay in aqueous phase | Uncertainty about whether assay really was in aqueous phase
Stability - Incubation Activity after incubation (1 hour at 25Β°C), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in aqueous phase Cyclopentyl Methyl Ether (CPME) 50% 104 106 % Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer Incubation: 7, Assay: 7 Incubation: 25Β°C, Assay: Room temperature 2 mM Ethyl 3-methyl-2-oxobutyrate Ethyl 3-methyl-2-hydroxybutyrate 0.2 mM NADPH Incubation: 100 rpm Water-incubated control (in %), assay in aqueous phase | Uncertainty about whether assay really was in aqueous phase
Stability - Incubation Activity after incubation (24 hours at 25Β°C), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in aqueous phase tert-Butyl Methyl Ether (MTBE) 50% 63 60 % Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer Incubation: 7, Assay: 7 Incubation: 25Β°C, Assay: Room temperature 2 mM Ethyl 3-methyl-2-oxobutyrate Ethyl 3-methyl-2-hydroxybutyrate 0.2 mM NADPH Incubation: 100 rpm Water-incubated control (in %), assay in aqueous phase | Uncertainty about whether assay really was in aqueous phase
Stability - Incubation Activity after incubation (5 hours at 25Β°C), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in aqueous phase tert-Butyl Methyl Ether (MTBE) 50% 83 82 % Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer Incubation: 7, Assay: 7 Incubation: 25Β°C, Assay: Room temperature 2 mM Ethyl 3-methyl-2-oxobutyrate Ethyl 3-methyl-2-hydroxybutyrate 0.2 mM NADPH Incubation: 100 rpm Water-incubated control (in %), assay in aqueous phase | Uncertainty about whether assay really was in aqueous phase
Stability - Incubation Activity after incubation (3 hours at 25Β°C), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in aqueous phase tert-Butyl Methyl Ether (MTBE) 50% 86 96 % Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer Incubation: 7, Assay: 7 Incubation: 25Β°C, Assay: Room temperature 2 mM Ethyl 3-methyl-2-oxobutyrate Ethyl 3-methyl-2-hydroxybutyrate 0.2 mM NADPH Incubation: 100 rpm Water-incubated control (in %), assay in aqueous phase | Uncertainty about whether assay really was in aqueous phase
Stability - Incubation Activity after incubation (1 hour at 25Β°C), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in aqueous phase tert-Butyl Methyl Ether (MTBE) 50% 104 9 % Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer Incubation: 7, Assay: 7 Incubation: 25Β°C, Assay: Room temperature 2 mM Ethyl 3-methyl-2-oxobutyrate Ethyl 3-methyl-2-hydroxybutyrate 0.2 mM NADPH Incubation: 100 rpm Water-incubated control (in %), assay in aqueous phase | Uncertainty about whether assay really was in aqueous phase
Stability - Incubation Activity after incubation (5 hours at room temperature), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in aqueous phase Dimethyl Sulfoxide (DMSO) 20% 86 83 % Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer Incubation: 7, Assay: 7 Incubation: Room temperature, Assay: Room temperature 2 mM Ethyl 3-methyl-2-oxobutyrate Ethyl 3-methyl-2-hydroxybutyrate 0.2 mM NADPH β€” Water-incubated control (in %), assay in aqueous phase | Uncertainty about whether assay really was in aqueous phase
Stability - Incubation Activity after incubation (5 hours at 25Β°C), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in aqueous phase Petroleum Ether 50% 83 70 % Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer Incubation: 7, Assay: 7 Incubation: 25Β°C, Assay: Room temperature 2 mM Ethyl 3-methyl-2-oxobutyrate Ethyl 3-methyl-2-hydroxybutyrate 0.2 mM NADPH Incubation: 100 rpm Water-incubated control (in %), assay in aqueous phase | Uncertainty about whether assay really was in aqueous phase
Stability - Incubation Activity after incubation (3 hours at 25Β°C), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in aqueous phase Petroleum Ether 50% 86 85 % Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer Incubation: 7, Assay: 7 Incubation: 25Β°C, Assay: Room temperature 2 mM Ethyl 3-methyl-2-oxobutyrate Ethyl 3-methyl-2-hydroxybutyrate 0.2 mM NADPH Incubation: 100 rpm Water-incubated control (in %), assay in aqueous phase | Uncertainty about whether assay really was in aqueous phase
Stability - Incubation Activity after incubation (1 hour at 25Β°C), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in aqueous phase Petroleum Ether 50% 104 93 % Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer Incubation: 7, Assay: 7 Incubation: 25Β°C, Assay: Room temperature 2 mM Ethyl 3-methyl-2-oxobutyrate Ethyl 3-methyl-2-hydroxybutyrate 0.2 mM NADPH Incubation: 100 rpm Water-incubated control (in %), assay in aqueous phase | Uncertainty about whether assay really was in aqueous phase
Stability - Incubation Activity after incubation (24 hours at 25Β°C), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in aqueous phase Ethyl Acetate 50% 63 62 % Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer Incubation: 7, Assay: 7 Incubation: 25Β°C, Assay: Room temperature 2 mM Ethyl 3-methyl-2-oxobutyrate Ethyl 3-methyl-2-hydroxybutyrate 0.2 mM NADPH Incubation: 100 rpm Water-incubated control (in %), assay in aqueous phase | Uncertainty about whether assay really was in aqueous phase
Stability - Incubation Activity after incubation (5 hours at 25Β°C), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in aqueous phase Ethyl Acetate 50% 83 78 % Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer Incubation: 7, Assay: 7 Incubation: 25Β°C, Assay: Room temperature 2 mM Ethyl 3-methyl-2-oxobutyrate Ethyl 3-methyl-2-hydroxybutyrate 0.2 mM NADPH Incubation: 100 rpm Water-incubated control (in %), assay in aqueous phase | Uncertainty about whether assay really was in aqueous phase
Stability - Incubation Activity after incubation (3 hours at 25Β°C), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in aqueous phase Ethyl Acetate 50% 86 89 % Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer Incubation: 7, Assay: 7 Incubation: 25Β°C, Assay: Room temperature 2 mM Ethyl 3-methyl-2-oxobutyrate Ethyl 3-methyl-2-hydroxybutyrate 0.2 mM NADPH Incubation: 100 rpm Water-incubated control (in %), assay in aqueous phase | Uncertainty about whether assay really was in aqueous phase
Stability - Incubation Activity after incubation (1 hour at 25Β°C), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in aqueous phase Ethyl Acetate 50% 104 94 % Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer Incubation: 7, Assay: 7 Incubation: 25Β°C, Assay: Room temperature 2 mM Ethyl 3-methyl-2-oxobutyrate Ethyl 3-methyl-2-hydroxybutyrate 0.2 mM NADPH Incubation: 100 rpm Water-incubated control (in %), assay in aqueous phase | Uncertainty about whether assay really was in aqueous phase
Stability - Incubation Activity after incubation (1 hour at room temperature), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in aqueous phase Methanol 20% 100 97 % Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer Incubation: 7, Assay: 7 Incubation: Room temperature, Assay: Room temperature 2 mM Ethyl 3-methyl-2-oxobutyrate Ethyl 3-methyl-2-hydroxybutyrate 0.2 mM NADPH β€” Water-incubated control (in %), assay in aqueous phase | Uncertainty about whether assay really was in aqueous phase
Stability - Incubation Activity after incubation (1 hour at room temperature), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in aqueous phase Ethanol 10% 100 113 % Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer Incubation: 7, Assay: 7 Incubation: Room temperature, Assay: Room temperature 2 mM Ethyl 3-methyl-2-oxobutyrate Ethyl 3-methyl-2-hydroxybutyrate 0.2 mM NADPH β€” Water-incubated control (in %), assay in aqueous phase | Uncertainty about whether assay really was in aqueous phase
Stability - Incubation Activity after incubation (24 hours at room temperature), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in aqueous phase Ethanol 5% 51 51 % Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer Incubation: 7, Assay: 7 Incubation: Room temperature, Assay: Room temperature 2 mM Ethyl 3-methyl-2-oxobutyrate Ethyl 3-methyl-2-hydroxybutyrate 0.2 mM NADPH β€” Water-incubated control (in %), assay in aqueous phase | Uncertainty about whether assay really was in aqueous phase
Stability - Incubation Activity after incubation (5 hours at room temperature), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in aqueous phase Ethanol 5% 86 91 % Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer Incubation: 7, Assay: 7 Incubation: Room temperature, Assay: Room temperature 2 mM Ethyl 3-methyl-2-oxobutyrate Ethyl 3-methyl-2-hydroxybutyrate 0.2 mM NADPH β€” Water-incubated control (in %), assay in aqueous phase | Uncertainty about whether assay really was in aqueous phase
Stability - Incubation Activity after incubation (3 hours at room temperature), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in aqueous phase Ethanol 5% 88 93 % Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer Incubation: 7, Assay: 7 Incubation: Room temperature, Assay: Room temperature 2 mM Ethyl 3-methyl-2-oxobutyrate Ethyl 3-methyl-2-hydroxybutyrate 0.2 mM NADPH β€” Water-incubated control (in %), assay in aqueous phase | Uncertainty about whether assay really was in aqueous phase
Stability - Incubation Activity after incubation (1 hour at room temperature), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in aqueous phase Ethanol 5% 100 59 % Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer Incubation: 7, Assay: 7 Incubation: Room temperature, Assay: Room temperature 2 mM Ethyl 3-methyl-2-oxobutyrate Ethyl 3-methyl-2-hydroxybutyrate 0.2 mM NADPH β€” Water-incubated control (in %), assay in aqueous phase | Uncertainty about whether assay really was in aqueous phase
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Visualization : Stability β€” Incubation

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Structure

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