Aldo-keto reductase from Kluyveromyces marxianus

Enzyme Description

Species
Kluyveromyces marxianus β†—
Extremophile
No
EC Number
1.1.1.21 β†—

Sequence

Length: 310 amino acids
MTNQKFFTLSNGNKIPAVAVVGTGTKWYKAEETDATFSQELTDIVKLSLDTVPGIVHIDAAETYKTYPELGAALKETKKPREEIFITDKFSSLHKISEDPKSALETALKKLGVDYVDLYLIHSPFFDKDLNIDLETAWKQLEELYKSGKAKNIGVSNFTVEDLKKVLAIAEIKPQVNQIEFSPFLQNQTPGIVEFSQKNDILLEAYSPLGPLQKKPADADQQPFYQYLKELSEKYKNTEAQVLLLWVYKRGILPVTTSAKIERIKQAQDIFSFDLTEEEVKKITDLGLQHEPVRLYWVDFYTKYNSEAQK
Shu-Fang Li et al. (2021) β€” Semirational engineering of an aldo-keto reductase KmAKR for overcoming trade-offs between catalytic activity and thermostability
Biotechnology and Bioengineering  Β· doi:10.1002/bit.27913 β†—  Β· Stability - Incubation
30 measurements ▼
Database ID
UniProt: A0ABX6EZM1 β†—
Sequence Annotation
Explicit - Provided
Protein Source
Recombinant, host bacterium Escherichia coli BL21 (DE3)

Experimental Data (30 measurements)

30 measurements β€” page 1 of 2
Mutation Property Assay Solvent Solvent Volume Aqueous ReferenceWT Reference Measured Value Units Solution pH Temperature Substrate(s) Product(s) Cofactor(s) Shaking Comments
W297H/Y296W/K29H/Y28A/T63M/A30P/T302S/S196C Stability - Incubation Activity after incubation (1 hour at 20Β°C), measured by HPLC in aquous conditions Ethanol 20% 100.0 β€” 43.2 % Incubation: 100 mM sodium phosphate buffer, Assay: 100 mM sodium phosphate buffer Incubation: 7, Assay: 7 Incubation: 20Β°C, Assay: 35Β°C 10 mM tert-butyl 6-chloro-(5S)-hydroxy-3-oxohexanoate tert-butyl 6-chloro-(3R,5S)-dihydroxyhexanoate 0.25 mM NADPH Assay: 800 rpm Classical aqueous control (in %)
W297H/Y296W/K29H/Y28A/T63M/A30P/T302S/S196C/K164E/S232A/S182H/Q266D Stability - Incubation Activity after incubation (1 hour at 20Β°C), measured by HPLC in aquous conditions Dimethylformamide (DMF) 20% 100.0 β€” 94.0 % Incubation: 100 mM sodium phosphate buffer, Assay: 100 mM sodium phosphate buffer Incubation: 7, Assay: 7 Incubation: 20Β°C, Assay: 35Β°C 10 mM tert-butyl 6-chloro-(5S)-hydroxy-3-oxohexanoate tert-butyl 6-chloro-(3R,5S)-dihydroxyhexanoate 0.25 mM NADPH Assay: 800 rpm Classical aqueous control (in %)
W297H/Y296W/K29H/Y28A/T63M/A30P/T302S/S196C/K164E/S232A/S182H/Q266D Stability - Incubation Activity after incubation (1 hour at 20Β°C), measured by HPLC in aquous conditions Isopropanol 20% 100.0 β€” 109.4 % Incubation: 100 mM sodium phosphate buffer, Assay: 100 mM sodium phosphate buffer Incubation: 7, Assay: 7 Incubation: 20Β°C, Assay: 35Β°C 10 mM tert-butyl 6-chloro-(5S)-hydroxy-3-oxohexanoate tert-butyl 6-chloro-(3R,5S)-dihydroxyhexanoate 0.25 mM NADPH Assay: 800 rpm Classical aqueous control (in %)
W297H/Y296W/K29H/Y28A/T63M/A30P/T302S/S196C/K164E/S232A/S182H/Q266D Stability - Incubation Activity after incubation (1 hour at 20Β°C), measured by HPLC in aquous conditions Methanol 20% 100.0 β€” 110.0 % Incubation: 100 mM sodium phosphate buffer, Assay: 100 mM sodium phosphate buffer Incubation: 7, Assay: 7 Incubation: 20Β°C, Assay: 35Β°C 10 mM tert-butyl 6-chloro-(5S)-hydroxy-3-oxohexanoate tert-butyl 6-chloro-(3R,5S)-dihydroxyhexanoate 0.25 mM NADPH Assay: 800 rpm Classical aqueous control (in %)
W297H/Y296W/K29H/Y28A/T63M/A30P/T302S/S196C/K164E/S232A/S182H/Q266D Stability - Incubation Activity after incubation (1 hour at 20Β°C), measured by HPLC in aquous conditions Dimethyl Sulfoxide (DMSO) 20% 100.0 β€” 108.8 % Incubation: 100 mM sodium phosphate buffer, Assay: 100 mM sodium phosphate buffer Incubation: 7, Assay: 7 Incubation: 20Β°C, Assay: 35Β°C 10 mM tert-butyl 6-chloro-(5S)-hydroxy-3-oxohexanoate tert-butyl 6-chloro-(3R,5S)-dihydroxyhexanoate 0.25 mM NADPH Assay: 800 rpm Classical aqueous control (in %)
W297H/Y296W/K29H/Y28A/T63M/A30P/T302S/S196C/K164E/S232A/S182H/Q266D Stability - Incubation Activity after incubation (1 hour at 20Β°C), measured by HPLC in aquous conditions Ethanol 20% 100.0 β€” 96.9 % Incubation: 100 mM sodium phosphate buffer, Assay: 100 mM sodium phosphate buffer Incubation: 7, Assay: 7 Incubation: 20Β°C, Assay: 35Β°C 10 mM tert-butyl 6-chloro-(5S)-hydroxy-3-oxohexanoate tert-butyl 6-chloro-(3R,5S)-dihydroxyhexanoate 0.25 mM NADPH Assay: 800 rpm Classical aqueous control (in %)
W297H/Y296W/K29H/Y28A/T63M/A30P/T302S/S196C/K164E/S232A/S182H/Q266D Stability - Incubation Activity after incubation (1 hour at 20Β°C), measured by HPLC in aquous conditions Acetonitrile 20% 100.0 β€” 96.2 % Incubation: 100 mM sodium phosphate buffer, Assay: 100 mM sodium phosphate buffer Incubation: 7, Assay: 7 Incubation: 20Β°C, Assay: 35Β°C 10 mM tert-butyl 6-chloro-(5S)-hydroxy-3-oxohexanoate tert-butyl 6-chloro-(3R,5S)-dihydroxyhexanoate 0.25 mM NADPH Assay: 800 rpm Classical aqueous control (in %)
W297H/Y296W/K29H/Y28A/T63M/A30P/T302S/S196C/K164E/S232A/S182H/Q266D Stability - Incubation Activity after incubation (1 hour at 20Β°C), measured by HPLC in aquous conditions p-Xylene 20% 100.0 β€” 118.9 % Incubation: 100 mM sodium phosphate buffer, Assay: 100 mM sodium phosphate buffer Incubation: 7, Assay: 7 Incubation: 20Β°C, Assay: 35Β°C 10 mM tert-butyl 6-chloro-(5S)-hydroxy-3-oxohexanoate tert-butyl 6-chloro-(3R,5S)-dihydroxyhexanoate 0.25 mM NADPH Assay: 800 rpm Classical aqueous control (in %)
W297H/Y296W/K29H/Y28A/T63M/A30P/T302S/S196C/K164E/S232A/S182H/Q266D Stability - Incubation Activity after incubation (1 hour at 20Β°C), measured by HPLC in aquous conditions Toluene 20% 100.0 β€” 111.3 % Incubation: 100 mM sodium phosphate buffer, Assay: 100 mM sodium phosphate buffer Incubation: 7, Assay: 7 Incubation: 20Β°C, Assay: 35Β°C 10 mM tert-butyl 6-chloro-(5S)-hydroxy-3-oxohexanoate tert-butyl 6-chloro-(3R,5S)-dihydroxyhexanoate 0.25 mM NADPH Assay: 800 rpm Classical aqueous control (in %)
W297H/Y296W/K29H/Y28A/T63M/A30P/T302S/S196C/K164E/S232A/S182H/Q266D Stability - Incubation Activity after incubation (1 hour at 20Β°C), measured by HPLC in aquous conditions 1-Octanol 20% 100.0 β€” 97.2 % Incubation: 100 mM sodium phosphate buffer, Assay: 100 mM sodium phosphate buffer Incubation: 7, Assay: 7 Incubation: 20Β°C, Assay: 35Β°C 10 mM tert-butyl 6-chloro-(5S)-hydroxy-3-oxohexanoate tert-butyl 6-chloro-(3R,5S)-dihydroxyhexanoate 0.25 mM NADPH Assay: 800 rpm Classical aqueous control (in %)
W297H/Y296W/K29H/Y28A/T63M/A30P/T302S/S196C/K164E/S232A/S182H/Q266D Stability - Incubation Activity after incubation (1 hour at 20Β°C), measured by HPLC in aquous conditions Isooctane 20% 100.0 β€” 108.5 % Incubation: 100 mM sodium phosphate buffer, Assay: 100 mM sodium phosphate buffer Incubation: 7, Assay: 7 Incubation: 20Β°C, Assay: 35Β°C 10 mM tert-butyl 6-chloro-(5S)-hydroxy-3-oxohexanoate tert-butyl 6-chloro-(3R,5S)-dihydroxyhexanoate 0.25 mM NADPH Assay: 800 rpm Classical aqueous control (in %)
W297H/Y296W/K29H/Y28A/T63M/A30P/T302S/S196C Stability - Incubation Activity after incubation (1 hour at 20Β°C), measured by HPLC in aquous conditions Dimethylformamide (DMF) 20% 100.0 β€” 94.3 % Incubation: 100 mM sodium phosphate buffer, Assay: 100 mM sodium phosphate buffer Incubation: 7, Assay: 7 Incubation: 20Β°C, Assay: 35Β°C 10 mM tert-butyl 6-chloro-(5S)-hydroxy-3-oxohexanoate tert-butyl 6-chloro-(3R,5S)-dihydroxyhexanoate 0.25 mM NADPH Assay: 800 rpm Classical aqueous control (in %)
W297H/Y296W/K29H/Y28A/T63M/A30P/T302S/S196C Stability - Incubation Activity after incubation (1 hour at 20Β°C), measured by HPLC in aquous conditions Isopropanol 20% 100.0 β€” 109.5 % Incubation: 100 mM sodium phosphate buffer, Assay: 100 mM sodium phosphate buffer Incubation: 7, Assay: 7 Incubation: 20Β°C, Assay: 35Β°C 10 mM tert-butyl 6-chloro-(5S)-hydroxy-3-oxohexanoate tert-butyl 6-chloro-(3R,5S)-dihydroxyhexanoate 0.25 mM NADPH Assay: 800 rpm Classical aqueous control (in %)
W297H/Y296W/K29H/Y28A/T63M/A30P/T302S/S196C Stability - Incubation Activity after incubation (1 hour at 20Β°C), measured by HPLC in aquous conditions Methanol 20% 100.0 β€” 103.7 % Incubation: 100 mM sodium phosphate buffer, Assay: 100 mM sodium phosphate buffer Incubation: 7, Assay: 7 Incubation: 20Β°C, Assay: 35Β°C 10 mM tert-butyl 6-chloro-(5S)-hydroxy-3-oxohexanoate tert-butyl 6-chloro-(3R,5S)-dihydroxyhexanoate 0.25 mM NADPH Assay: 800 rpm Classical aqueous control (in %)
W297H/Y296W/K29H/Y28A/T63M/A30P/T302S/S196C Stability - Incubation Activity after incubation (1 hour at 20Β°C), measured by HPLC in aquous conditions Dimethyl Sulfoxide (DMSO) 20% 100.0 β€” 105.2 % Incubation: 100 mM sodium phosphate buffer, Assay: 100 mM sodium phosphate buffer Incubation: 7, Assay: 7 Incubation: 20Β°C, Assay: 35Β°C 10 mM tert-butyl 6-chloro-(5S)-hydroxy-3-oxohexanoate tert-butyl 6-chloro-(3R,5S)-dihydroxyhexanoate 0.25 mM NADPH Assay: 800 rpm Classical aqueous control (in %)
W297H/Y296W/K29H/Y28A/T63M Stability - Incubation Activity after incubation (1 hour at 20Β°C), measured by HPLC in aquous conditions Isooctane 20% 100.0 β€” 56.2 % Incubation: 100 mM sodium phosphate buffer, Assay: 100 mM sodium phosphate buffer Incubation: 7, Assay: 7 Incubation: 20Β°C, Assay: 35Β°C 10 mM tert-butyl 6-chloro-(5S)-hydroxy-3-oxohexanoate tert-butyl 6-chloro-(3R,5S)-dihydroxyhexanoate 0.25 mM NADPH Assay: 800 rpm Classical aqueous control (in %)
W297H/Y296W/K29H/Y28A/T63M/A30P/T302S/S196C Stability - Incubation Activity after incubation (1 hour at 20Β°C), measured by HPLC in aquous conditions Acetonitrile 20% 100.0 β€” 84.2 % Incubation: 100 mM sodium phosphate buffer, Assay: 100 mM sodium phosphate buffer Incubation: 7, Assay: 7 Incubation: 20Β°C, Assay: 35Β°C 10 mM tert-butyl 6-chloro-(5S)-hydroxy-3-oxohexanoate tert-butyl 6-chloro-(3R,5S)-dihydroxyhexanoate 0.25 mM NADPH Assay: 800 rpm Classical aqueous control (in %)
W297H/Y296W/K29H/Y28A/T63M/A30P/T302S/S196C Stability - Incubation Activity after incubation (1 hour at 20Β°C), measured by HPLC in aquous conditions p-Xylene 20% 100.0 β€” 104.9 % Incubation: 100 mM sodium phosphate buffer, Assay: 100 mM sodium phosphate buffer Incubation: 7, Assay: 7 Incubation: 20Β°C, Assay: 35Β°C 10 mM tert-butyl 6-chloro-(5S)-hydroxy-3-oxohexanoate tert-butyl 6-chloro-(3R,5S)-dihydroxyhexanoate 0.25 mM NADPH Assay: 800 rpm Classical aqueous control (in %)
W297H/Y296W/K29H/Y28A/T63M/A30P/T302S/S196C Stability - Incubation Activity after incubation (1 hour at 20Β°C), measured by HPLC in aquous conditions Toluene 20% 100.0 β€” 98.9 % Incubation: 100 mM sodium phosphate buffer, Assay: 100 mM sodium phosphate buffer Incubation: 7, Assay: 7 Incubation: 20Β°C, Assay: 35Β°C 10 mM tert-butyl 6-chloro-(5S)-hydroxy-3-oxohexanoate tert-butyl 6-chloro-(3R,5S)-dihydroxyhexanoate 0.25 mM NADPH Assay: 800 rpm Classical aqueous control (in %)
W297H/Y296W/K29H/Y28A/T63M/A30P/T302S/S196C Stability - Incubation Activity after incubation (1 hour at 20Β°C), measured by HPLC in aquous conditions 1-Octanol 20% 100.0 β€” 64.7 % Incubation: 100 mM sodium phosphate buffer, Assay: 100 mM sodium phosphate buffer Incubation: 7, Assay: 7 Incubation: 20Β°C, Assay: 35Β°C 10 mM tert-butyl 6-chloro-(5S)-hydroxy-3-oxohexanoate tert-butyl 6-chloro-(3R,5S)-dihydroxyhexanoate 0.25 mM NADPH Assay: 800 rpm Classical aqueous control (in %)
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Mutations in this dataset (3)

W297H/Y296W/K29H/Y28A/T63M W297H/Y296W/K29H/Y28A/T63M/A30P/T302S/S196C W297H/Y296W/K29H/Y28A/T63M/A30P/T302S/S196C/K164E/S232A/S182H/Q266D

Visualization : Stability β€” Incubation

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

Structure

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