Aldo-keto reductase (gene KdAKR) from Kluyvermyces dobzhanskii

Enzyme Description

Species
Kluyvermyces dobzhanskii β†—
Extremophile
No
EC Number
1.1.1.2 β†—

Sequence

Length: 310 amino acids
MTTQQFFTLSNGNKIPAVAIVGTGTAWYKSEETDATFSQDLVNIVKETLDTVPGVVHLDAAEIYRTYPELGAALKDTKKPREEIFITDKYSTLLQLSENPTVALESSLKKLGVDYVDLYLIHSPIIDKNGTIDIETAWKHLEALYKSGKAKNIGVSNFTVEDLEKLLAVAEIKPQVNQIEFSPFLQNQTPGIVEFSQKNNILLEAYSPLGPLQKRPADSDKIGFYSYIAELTKKYSKSEAQVILLWVYKRGILPVTTSSKIERVQQAQDIFSFDLTDEEVKKITQLGLEQPALRLYWADVYTKYNSEAQK
Chen Dai et al. (2024) β€” Computer-aided design to enhance the stability of aldo-keto reductase Kd AKR
Biotechnology Journal  Β· doi:10.1002/biot.202300637 β†—  Β· Stability - Incubation
18 measurements ▼
Database ID
β€”
Sequence Annotation
Explicit - Provided
Protein Source
Recombinant, host bacterium Escherichia coli BL21 (DE3)

Experimental Data (18 measurements)

18 measurements
Mutation Property Assay Solvent Solvent Volume Aqueous ReferenceWT Reference Measured Value Units Solution pH Temperature Substrate(s) Product(s) Cofactor(s) Shaking Comments
Y28A/V45L/L58I/I63L/S196C/G223P/T232A/V264I/Y296W/W297H Stability - Incubation Activity after incubation (1 hour at 20Β°C), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in aqueous phase Ethanol 25% 100.0 β€” 93.0 % Incubation: ?, Assay: 100 mM potassium phosphate buffer Incubation: ?, Assay: 7 Incubation: 20Β°C, Assay: 35Β°c 10 mM tert-Butyl (5S)-6-chloro-5-hydroxy-3-oxohexanoate t-butyl 6-chloro- (3R,5S)-dihydroxyhexanoate 1 mM NADPH β€” Non-incubated control (in %), assay in aqueous phase | Uncertainty about whether assay really was in aqueous phase
Y28A/V45L/L58I/I63L/S196C/G223P/T232A/V264I/Y296W/W297H Stability - Incubation Activity after incubation (1 hour at 20Β°C), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in aqueous phase p-Xylene 25% 100.0 β€” 102.3 % Incubation: ?, Assay: 100 mM potassium phosphate buffer Incubation: ?, Assay: 7 Incubation: 20Β°C, Assay: 35Β°c 10 mM tert-Butyl (5S)-6-chloro-5-hydroxy-3-oxohexanoate t-butyl 6-chloro- (3R,5S)-dihydroxyhexanoate 1 mM NADPH β€” Non-incubated control (in %), assay in aqueous phase | Uncertainty about whether assay really was in aqueous phase
Y28A/L58I/I63L/G223P/Y296W/W297H Stability - Incubation Activity after incubation (1 hour at 20Β°C), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in aqueous phase p-Xylene 25% 100.0 β€” 101.2 % Incubation: ?, Assay: 100 mM potassium phosphate buffer Incubation: ?, Assay: 7 Incubation: 20Β°C, Assay: 35Β°c 10 mM tert-Butyl (5S)-6-chloro-5-hydroxy-3-oxohexanoate t-butyl 6-chloro- (3R,5S)-dihydroxyhexanoate 1 mM NADPH β€” Non-incubated control (in %), assay in aqueous phase | Uncertainty about whether assay really was in aqueous phase
Y28A/V45L/L58I/I63L/S196C/G223P/T232A/V264I/Y296W/W297H Stability - Incubation Activity after incubation (1 hour at 20Β°C), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in aqueous phase 1-Octanol 25% 100.0 β€” 96.1 % Incubation: ?, Assay: 100 mM potassium phosphate buffer Incubation: ?, Assay: 7 Incubation: 20Β°C, Assay: 35Β°c 10 mM tert-Butyl (5S)-6-chloro-5-hydroxy-3-oxohexanoate t-butyl 6-chloro- (3R,5S)-dihydroxyhexanoate 1 mM NADPH β€” Non-incubated control (in %), assay in aqueous phase | Uncertainty about whether assay really was in aqueous phase
Y28A/L58I/I63L/G223P/Y296W/W297H Stability - Incubation Activity after incubation (1 hour at 20Β°C), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in aqueous phase 1-Octanol 25% 100.0 β€” 27.5 % Incubation: ?, Assay: 100 mM potassium phosphate buffer Incubation: ?, Assay: 7 Incubation: 20Β°C, Assay: 35Β°c 10 mM tert-Butyl (5S)-6-chloro-5-hydroxy-3-oxohexanoate t-butyl 6-chloro- (3R,5S)-dihydroxyhexanoate 1 mM NADPH β€” Non-incubated control (in %), assay in aqueous phase | Uncertainty about whether assay really was in aqueous phase
Y28A/V45L/L58I/I63L/S196C/G223P/T232A/V264I/Y296W/W297H Stability - Incubation Activity after incubation (1 hour at 20Β°C), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in aqueous phase tert-Butyl Methyl Ether (MTBE) 25% 100.0 β€” 103.8 % Incubation: ?, Assay: 100 mM potassium phosphate buffer Incubation: ?, Assay: 7 Incubation: 20Β°C, Assay: 35Β°c 10 mM tert-Butyl (5S)-6-chloro-5-hydroxy-3-oxohexanoate t-butyl 6-chloro- (3R,5S)-dihydroxyhexanoate 1 mM NADPH β€” Non-incubated control (in %), assay in aqueous phase | Uncertainty about whether assay really was in aqueous phase
Y28A/L58I/I63L/G223P/Y296W/W297H Stability - Incubation Activity after incubation (1 hour at 20Β°C), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in aqueous phase tert-Butyl Methyl Ether (MTBE) 25% 100.0 β€” 42.1 % Incubation: ?, Assay: 100 mM potassium phosphate buffer Incubation: ?, Assay: 7 Incubation: 20Β°C, Assay: 35Β°c 10 mM tert-Butyl (5S)-6-chloro-5-hydroxy-3-oxohexanoate t-butyl 6-chloro- (3R,5S)-dihydroxyhexanoate 1 mM NADPH β€” Non-incubated control (in %), assay in aqueous phase | Uncertainty about whether assay really was in aqueous phase
Y28A/V45L/L58I/I63L/S196C/G223P/T232A/V264I/Y296W/W297H Stability - Incubation Activity after incubation (1 hour at 20Β°C), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in aqueous phase Isopropanol 25% 100.0 β€” 56.0 % Incubation: ?, Assay: 100 mM potassium phosphate buffer Incubation: ?, Assay: 7 Incubation: 20Β°C, Assay: 35Β°c 10 mM tert-Butyl (5S)-6-chloro-5-hydroxy-3-oxohexanoate t-butyl 6-chloro- (3R,5S)-dihydroxyhexanoate 1 mM NADPH β€” Non-incubated control (in %), assay in aqueous phase | Uncertainty about whether assay really was in aqueous phase
Y28A/L58I/I63L/G223P/Y296W/W297H Stability - Incubation Activity after incubation (1 hour at 20Β°C), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in aqueous phase Isopropanol 25% 100.0 β€” 24.7 % Incubation: ?, Assay: 100 mM potassium phosphate buffer Incubation: ?, Assay: 7 Incubation: 20Β°C, Assay: 35Β°c 10 mM tert-Butyl (5S)-6-chloro-5-hydroxy-3-oxohexanoate t-butyl 6-chloro- (3R,5S)-dihydroxyhexanoate 1 mM NADPH β€” Non-incubated control (in %), assay in aqueous phase | Uncertainty about whether assay really was in aqueous phase
Y28A/L58I/I63L/G223P/Y296W/W297H Stability - Incubation Activity after incubation (1 hour at 20Β°C), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in aqueous phase Dimethyl Sulfoxide (DMSO) 25% 100.0 β€” 43.2 % Incubation: ?, Assay: 100 mM potassium phosphate buffer Incubation: ?, Assay: 7 Incubation: 20Β°C, Assay: 35Β°c 10 mM tert-Butyl (5S)-6-chloro-5-hydroxy-3-oxohexanoate t-butyl 6-chloro- (3R,5S)-dihydroxyhexanoate 1 mM NADPH β€” Non-incubated control (in %), assay in aqueous phase | Uncertainty about whether assay really was in aqueous phase
Y28A/L58I/I63L/G223P/Y296W/W297H Stability - Incubation Activity after incubation (1 hour at 20Β°C), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in aqueous phase Ethanol 25% 100.0 β€” 24.9 % Incubation: ?, Assay: 100 mM potassium phosphate buffer Incubation: ?, Assay: 7 Incubation: 20Β°C, Assay: 35Β°c 10 mM tert-Butyl (5S)-6-chloro-5-hydroxy-3-oxohexanoate t-butyl 6-chloro- (3R,5S)-dihydroxyhexanoate 1 mM NADPH β€” Non-incubated control (in %), assay in aqueous phase | Uncertainty about whether assay really was in aqueous phase
Y28A/V45L/L58I/I63L/S196C/G223P/T232A/V264I/Y296W/W297H Stability - Incubation Activity after incubation (1 hour at 20Β°C), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in aqueous phase Acetonitrile 25% 100.0 β€” 78.6 % Incubation: ?, Assay: 100 mM potassium phosphate buffer Incubation: ?, Assay: 7 Incubation: 20Β°C, Assay: 35Β°c 10 mM tert-Butyl (5S)-6-chloro-5-hydroxy-3-oxohexanoate t-butyl 6-chloro- (3R,5S)-dihydroxyhexanoate 1 mM NADPH β€” Non-incubated control (in %), assay in aqueous phase | Uncertainty about whether assay really was in aqueous phase
Y28A/L58I/I63L/G223P/Y296W/W297H Stability - Incubation Activity after incubation (1 hour at 20Β°C), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in aqueous phase Acetonitrile 25% 100.0 β€” 28.8 % Incubation: ?, Assay: 100 mM potassium phosphate buffer Incubation: ?, Assay: 7 Incubation: 20Β°C, Assay: 35Β°c 10 mM tert-Butyl (5S)-6-chloro-5-hydroxy-3-oxohexanoate t-butyl 6-chloro- (3R,5S)-dihydroxyhexanoate 1 mM NADPH β€” Non-incubated control (in %), assay in aqueous phase | Uncertainty about whether assay really was in aqueous phase
Y28A/V45L/L58I/I63L/S196C/G223P/T232A/V264I/Y296W/W297H Stability - Incubation Activity after incubation (1 hour at 20Β°C), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in aqueous phase Methanol 25% 100.0 β€” 91.5 % Incubation: ?, Assay: 100 mM potassium phosphate buffer Incubation: ?, Assay: 7 Incubation: 20Β°C, Assay: 35Β°c 10 mM tert-Butyl (5S)-6-chloro-5-hydroxy-3-oxohexanoate t-butyl 6-chloro- (3R,5S)-dihydroxyhexanoate 1 mM NADPH β€” Non-incubated control (in %), assay in aqueous phase | Uncertainty about whether assay really was in aqueous phase
Y28A/L58I/I63L/G223P/Y296W/W297H Stability - Incubation Activity after incubation (1 hour at 20Β°C), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in aqueous phase Methanol 25% 100.0 β€” 85.8 % Incubation: ?, Assay: 100 mM potassium phosphate buffer Incubation: ?, Assay: 7 Incubation: 20Β°C, Assay: 35Β°c 10 mM tert-Butyl (5S)-6-chloro-5-hydroxy-3-oxohexanoate t-butyl 6-chloro- (3R,5S)-dihydroxyhexanoate 1 mM NADPH β€” Non-incubated control (in %), assay in aqueous phase | Uncertainty about whether assay really was in aqueous phase
Y28A/V45L/L58I/I63L/S196C/G223P/T232A/V264I/Y296W/W297H Stability - Incubation Activity after incubation (1 hour at 20Β°C), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in aqueous phase Dimethylformamide (DMF) 25% 100.0 β€” 103.3 % Incubation: ?, Assay: 100 mM potassium phosphate buffer Incubation: ?, Assay: 7 Incubation: 20Β°C, Assay: 35Β°c 10 mM tert-Butyl (5S)-6-chloro-5-hydroxy-3-oxohexanoate t-butyl 6-chloro- (3R,5S)-dihydroxyhexanoate 1 mM NADPH β€” Non-incubated control (in %), assay in aqueous phase | Uncertainty about whether assay really was in aqueous phase
Y28A/L58I/I63L/G223P/Y296W/W297H Stability - Incubation Activity after incubation (1 hour at 20Β°C), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in aqueous phase Dimethylformamide (DMF) 25% 100.0 β€” 39.0 % Incubation: ?, Assay: 100 mM potassium phosphate buffer Incubation: ?, Assay: 7 Incubation: 20Β°C, Assay: 35Β°c 10 mM tert-Butyl (5S)-6-chloro-5-hydroxy-3-oxohexanoate t-butyl 6-chloro- (3R,5S)-dihydroxyhexanoate 1 mM NADPH β€” Non-incubated control (in %), assay in aqueous phase | Uncertainty about whether assay really was in aqueous phase
Y28A/V45L/L58I/I63L/S196C/G223P/T232A/V264I/Y296W/W297H Stability - Incubation Activity after incubation (1 hour at 20Β°C), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in aqueous phase Dimethyl Sulfoxide (DMSO) 25% 100.0 β€” 45.5 % Incubation: ?, Assay: 100 mM potassium phosphate buffer Incubation: ?, Assay: 7 Incubation: 20Β°C, Assay: 35Β°c 10 mM tert-Butyl (5S)-6-chloro-5-hydroxy-3-oxohexanoate t-butyl 6-chloro- (3R,5S)-dihydroxyhexanoate 1 mM NADPH β€” Non-incubated control (in %), assay in aqueous phase | Uncertainty about whether assay really was in aqueous phase

Mutations in this dataset (2)

Y28A/L58I/I63L/G223P/Y296W/W297H Y28A/V45L/L58I/I63L/S196C/G223P/T232A/V264I/Y296W/W297H

Visualization : Stability β€” Incubation

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

Structure

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