Diaryl alcohol dehydrogenase (gene KpADH) from Kluyveromyces polyspora

Enzyme Description

Extremophile
No
EC Number

Sequence

Length: 342 amino acids
MSVLISGASGYIAKHIVRVLLEQNYKVIGTVRSQDKADKLLKQYNNPNLSYEIVPEIANLDAFDDIFKKHGKEIKYVIHAASPVNFGAKDLEKDLVIPAINGTKNMFEAIKKYAPDTVERVVMTASYASIMTPHRQNDPTLTLDEETWNPVTEENAYENVFTAYCASKTFAEKEAWKFVKENSDAVKFKLTTIHPSFVFGPQNFDEDVTKKLNETCEIINGLLHAPFDTKVEKTHFSQFIDVRDVAKTHVLGFQKDELINQRLLLCNGAFSQQDIVNVFNEDFPELKGQFPPEDKDTDLNKGVTGCKIDNEKTKKLLAFEFTPFHKTIHDTVYQILHKEGRV
Lu Zhang et al. (2024) β€” Engineering diaryl alcohol dehydrogenase Kp ADH reveals importance of retaining hydration shell in organic solvent tolerance
Protein Science  Β· doi:10.1002/pro.4933 β†—  Β· Activity - Classical Activity - Michaelis-Menten
139 measurements
Database ID
UniProt: A7TM80 β†—
Sequence Annotation
Explicit - Provided
Protein Source
Recombinant, host bacterium Escherichia coli BL21 (DE3)

Experimental Data (139 measurements)

139 measurements β€” page 3 of 7
Mutation Property Assay Solvent Solvent Volume Aqueous ReferenceWT Reference Measured Value Units Solution pH Temperature Substrate(s) Product(s) Cofactor(s) Shaking Comments
WT Activity - Classical Activity measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm), with the assay with 5% Ethanol as control Ethanol 15% 100.0 β€” 7.4 % 100 mM sodium phosphate buffer 6 30Β°C ? mM Methyl 1-Boc-4-hydroxypiperidine-2-carboxylate Methyl N-Boc-4-oxopiperidine-2-carboxylate ? mM NADPH β€” Classical aqueous control (in %) | Reference measured with 5% ethanol
V231D Activity - Classical Activity measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm), with the assay with 5% Ethanol as control Methanol 2.5% 100.0 82.3 88.1 % 100 mM sodium phosphate buffer 6 30Β°C ? mM 2-(4-Chlorobenzoyl)pyridine CPMK-alcohol ? mM NADPH β€” Classical aqueous control (in %) | Reference measured with 5% ethanol
V231D Activity - Classical Activity measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm), with the assay with 5% Ethanol as control Methanol 5% 100.0 63.1 80.8 % 100 mM sodium phosphate buffer 6 30Β°C ? mM 2-(4-Chlorobenzoyl)pyridine CPMK-alcohol ? mM NADPH β€” Classical aqueous control (in %) | Reference measured with 5% ethanol
V231D Activity - Classical Activity measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm), with the assay with 5% Ethanol as control Methanol 7.5% 100.0 38.2 62.1 % 100 mM sodium phosphate buffer 6 30Β°C ? mM 2-(4-Chlorobenzoyl)pyridine CPMK-alcohol ? mM NADPH β€” Classical aqueous control (in %) | Reference measured with 5% ethanol
V231D Activity - Classical Activity measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm), with the assay with 5% Ethanol as control Methanol 10% 100.0 32.6 41.2 % 100 mM sodium phosphate buffer 6 30Β°C ? mM 2-(4-Chlorobenzoyl)pyridine CPMK-alcohol ? mM NADPH β€” Classical aqueous control (in %) | Reference measured with 5% ethanol
V231D Activity - Classical Activity measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm), with the assay with 5% Ethanol as control Methanol 12.5% 100.0 26.4 33.9 % 100 mM sodium phosphate buffer 6 30Β°C ? mM 2-(4-Chlorobenzoyl)pyridine CPMK-alcohol ? mM NADPH β€” Classical aqueous control (in %) | Reference measured with 5% ethanol
V231D Activity - Classical Activity measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm), with the assay with 5% Ethanol as control Methanol 15% 100.0 18.7 22.0 % 100 mM sodium phosphate buffer 6 30Β°C ? mM 2-(4-Chlorobenzoyl)pyridine CPMK-alcohol ? mM NADPH β€” Classical aqueous control (in %) | Reference measured with 5% ethanol
S237G Activity - Classical Activity measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm), with the assay with 5% Ethanol as control Methanol 2.5% 100.0 82.3 75.1 % 100 mM sodium phosphate buffer 6 30Β°C ? mM 2-(4-Chlorobenzoyl)pyridine CPMK-alcohol ? mM NADPH β€” Classical aqueous control (in %) | Reference measured with 5% ethanol
S237G Activity - Classical Activity measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm), with the assay with 5% Ethanol as control Methanol 5% 100.0 63.1 70.6 % 100 mM sodium phosphate buffer 6 30Β°C ? mM 2-(4-Chlorobenzoyl)pyridine CPMK-alcohol ? mM NADPH β€” Classical aqueous control (in %) | Reference measured with 5% ethanol
S237G Activity - Classical Activity measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm), with the assay with 5% Ethanol as control Methanol 7.5% 100.0 38.2 43.5 % 100 mM sodium phosphate buffer 6 30Β°C ? mM 2-(4-Chlorobenzoyl)pyridine CPMK-alcohol ? mM NADPH β€” Classical aqueous control (in %) | Reference measured with 5% ethanol
S237G Activity - Classical Activity measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm), with the assay with 5% Ethanol as control Methanol 10% 100.0 32.6 32.8 % 100 mM sodium phosphate buffer 6 30Β°C ? mM 2-(4-Chlorobenzoyl)pyridine CPMK-alcohol ? mM NADPH β€” Classical aqueous control (in %) | Reference measured with 5% ethanol
S237G Activity - Classical Activity measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm), with the assay with 5% Ethanol as control Methanol 12.5% 100.0 26.4 27.1 % 100 mM sodium phosphate buffer 6 30Β°C ? mM 2-(4-Chlorobenzoyl)pyridine CPMK-alcohol ? mM NADPH β€” Classical aqueous control (in %) | Reference measured with 5% ethanol
S237G Activity - Classical Activity measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm), with the assay with 5% Ethanol as control Methanol 15% 100.0 18.7 18.1 % 100 mM sodium phosphate buffer 6 30Β°C ? mM 2-(4-Chlorobenzoyl)pyridine CPMK-alcohol ? mM NADPH β€” Classical aqueous control (in %) | Reference measured with 5% ethanol
V231D Activity - Classical Activity measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm), with the assay with 5% Ethanol as control Ethanol 7.5% 100.0 69.1 91.7 % 100 mM sodium phosphate buffer 6 30Β°C ? mM 2-(4-Chlorobenzoyl)pyridine CPMK-alcohol ? mM NADPH β€” Classical aqueous control (in %) | Reference measured with 5% ethanol
V231D Activity - Classical Activity measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm), with the assay with 5% Ethanol as control Ethanol 10% 100.0 49.4 69.8 % 100 mM sodium phosphate buffer 6 30Β°C ? mM 2-(4-Chlorobenzoyl)pyridine CPMK-alcohol ? mM NADPH β€” Classical aqueous control (in %) | Reference measured with 5% ethanol
V231D Activity - Classical Activity measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm), with the assay with 5% Ethanol as control Ethanol 12.5% 100.0 33.1 54.0 % 100 mM sodium phosphate buffer 6 30Β°C ? mM 2-(4-Chlorobenzoyl)pyridine CPMK-alcohol ? mM NADPH β€” Classical aqueous control (in %) | Reference measured with 5% ethanol
V231D Activity - Classical Activity measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm), with the assay with 5% Ethanol as control Ethanol 15% 100.0 23.0 37.7 % 100 mM sodium phosphate buffer 6 30Β°C ? mM 2-(4-Chlorobenzoyl)pyridine CPMK-alcohol ? mM NADPH β€” Classical aqueous control (in %) | Reference measured with 5% ethanol
V231D Activity - Classical Activity measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm), with the assay with 5% Ethanol as control Ethanol 17.5% 100.0 19.7 19.7 % 100 mM sodium phosphate buffer 6 30Β°C ? mM 2-(4-Chlorobenzoyl)pyridine CPMK-alcohol ? mM NADPH β€” Classical aqueous control (in %) | Reference measured with 5% ethanol
V231D Activity - Classical Activity measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm), with the assay with 5% Ethanol as control Ethanol 20% 100.0 13.0 11.9 % 100 mM sodium phosphate buffer 6 30Β°C ? mM 2-(4-Chlorobenzoyl)pyridine CPMK-alcohol ? mM NADPH β€” Classical aqueous control (in %) | Reference measured with 5% ethanol
S237G Activity - Classical Activity measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm), with the assay with 5% Ethanol as control Ethanol 7.5% 100.0 69.1 86.1 % 100 mM sodium phosphate buffer 6 30Β°C ? mM 2-(4-Chlorobenzoyl)pyridine CPMK-alcohol ? mM NADPH β€” Classical aqueous control (in %) | Reference measured with 5% ethanol
1 2 3 4 … 7

Mutations in this dataset (3)

WT V231D S237G

Visualization : Activity β€” Classical

One bar per measurement. Colour = solvent, shade = solvent volume. β€” β€” β€” Reference value. Hover for details.

Mutation Effect

Mutation impact on enzyme stability and function in the presence of organic solvent: comparison of wild-type and mutant values in identical conditions.

Visualization : Activity β€” Michaelis-Menten

One bar per measurement. Colour = solvent, shade = solvent volume. β€” β€” β€” Reference value. Hover for details.

Mutation Effect β€” Michaelis-Menten

Mutation impact on kinetic parameters in the presence of organic solvent: comparison of wild-type and mutant values in identical conditions.

Structure

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