Diaryl alcohol dehydrogenase (gene KpADH) from Kluyveromyces polyspora

Enzyme Description

Species

Extremophile

EC Number

1.1.1.1 ↗

Sequence

Length: 342 amino acids

MSVLISGASGYIAKHIVRVLLEQNYKVIGTVRSQDKADKLLKQYNNPNLSYEIVPEIANLDAFDDIFKKHGKEIKYVIHAASPVNFGAKDLEKDLVIPAINGTKNMFEAIKKYAPDTVERVVMTASYASIMTPHRQNDPTLTLDEETWNPVTEENAYENVFTAYCASKTFAEKEAWKFVKENSDAVKFKLTTIHPSFVFGPQNFDEDVTKKLNETCEIINGLLHAPFDTKVEKTHFSQFIDVRDVAKTHVLGFQKDELINQRLLLCNGAFSQQDIVNVFNEDFPELKGQFPPEDKDTDLNKGVTGCKIDNEKTKKLLAFEFTPFHKTIHDTVYQILHKEGRV

Lu Zhang et al. (2024) — Engineering diaryl alcohol dehydrogenase Kp ADH reveals importance of retaining hydration shell in organic solvent tolerance

Protein Science · doi:10.1002/pro.4933 ↗ · Activity - Classical Activity - Michaelis-Menten

139 measurements ▼

Database ID

UniProt: A7TM80 ↗

Sequence Annotation

Explicit - Provided

Protein Source

Recombinant, host bacterium Escherichia coli BL21 (DE3)

Experimental Data (139 measurements)

139 measurements — page 7 of 7

Mutation	Property	Assay	Solvent	Solvent Volume	Aqueous Reference	WT Reference	Measured Value	Units	Solution	pH	Temperature	Substrate(s)	Product(s)	Cofactor(s)	Shaking	Comments
S237G	Activity - Classical	Activity measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm), with the assay with 5% Ethanol as control	Ethanol	15%	100.0	7.4	22.3	%	100 mM sodium phosphate buffer	6	30°C	? mM Methyl 1-Boc-4-hydroxypiperidine-2-carboxylate	Methyl N-Boc-4-oxopiperidine-2-carboxylate	? mM NADPH	—	Classical aqueous control (in %) \| Reference measured with 5% ethanol
V231D	Activity - Michaelis-Menten	Km (mM), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm), with the assay with 5% Ethanol as control	Ethanol	5%	—	2.29	22.74	mM	100 mM sodium phosphate buffer	6	30°C	2-(4-Chlorobenzoyl)pyridine	CPMK-alcohol	? mM NADPH	—	Wild-type control with organic solvent (in mM) \| Control is not mutant value in aqueous phase, but WT value in organic solvent
V231D	Activity - Michaelis-Menten	kcat (1/s), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm), with the assay with 5% Ethanol as control	Ethanol	5%	—	32.05	12.81	1/s	100 mM sodium phosphate buffer	6	30°C	2-(4-Chlorobenzoyl)pyridine	CPMK-alcohol	? mM NADPH	—	Wild-type control with organic solvent (in 1/s) \| Control is not mutant value in aqueous phase, but WT value in organic solvent
V231D	Activity - Michaelis-Menten	kcat/Km (s^-1 mM^-1), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm), with the assay with 5% Ethanol as control	Ethanol	5%	—	14.0	0.56	s^-1 * mM^-1	100 mM sodium phosphate buffer	6	30°C	2-(4-Chlorobenzoyl)pyridine	CPMK-alcohol	? mM NADPH	—	Wild-type control with organic solvent (in s^-1 * mM^-1) \| Control is not mutant value in aqueous phase, but WT value in organic solvent
S237G	Activity - Michaelis-Menten	Km (mM), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm), with the assay with 5% Ethanol as control	Ethanol	5%	—	2.29	1.07	mM	100 mM sodium phosphate buffer	6	30°C	2-(4-Chlorobenzoyl)pyridine	CPMK-alcohol	? mM NADPH	—	Wild-type control with organic solvent (in mM) \| Control is not mutant value in aqueous phase, but WT value in organic solvent
S237G	Activity - Michaelis-Menten	kcat (1/s), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm), with the assay with 5% Ethanol as control	Ethanol	5%	—	32.05	97.7	1/s	100 mM sodium phosphate buffer	6	30°C	2-(4-Chlorobenzoyl)pyridine	CPMK-alcohol	? mM NADPH	—	Wild-type control with organic solvent (in 1/s) \| Control is not mutant value in aqueous phase, but WT value in organic solvent
S237G	Activity - Michaelis-Menten	kcat/Km (s^-1 mM^-1), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm), with the assay with 5% Ethanol as control	Ethanol	5%	—	14.0	91.56	s^-1 * mM^-1	100 mM sodium phosphate buffer	6	30°C	2-(4-Chlorobenzoyl)pyridine	CPMK-alcohol	? mM NADPH	—	Wild-type control with organic solvent (in s^-1 * mM^-1) \| Control is not mutant value in aqueous phase, but WT value in organic solvent
V231D	Activity - Michaelis-Menten	Km (mM), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm), with the assay with 5% Ethanol as control	Ethanol	10%	—	3.13	44.23	mM	100 mM sodium phosphate buffer	6	30°C	2-(4-Chlorobenzoyl)pyridine	CPMK-alcohol	? mM NADPH	—	Wild-type control with organic solvent (in mM) \| Control is not mutant value in aqueous phase, but WT value in organic solvent
V231D	Activity - Michaelis-Menten	kcat (1/s), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm), with the assay with 5% Ethanol as control	Ethanol	10%	—	20.82	8.68	1/s	100 mM sodium phosphate buffer	6	30°C	2-(4-Chlorobenzoyl)pyridine	CPMK-alcohol	? mM NADPH	—	Wild-type control with organic solvent (in 1/s) \| Control is not mutant value in aqueous phase, but WT value in organic solvent
V231D	Activity - Michaelis-Menten	kcat/Km (s^-1 mM^-1), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm), with the assay with 5% Ethanol as control	Ethanol	10%	—	6.65	0.196	s^-1 * mM^-1	100 mM sodium phosphate buffer	6	30°C	2-(4-Chlorobenzoyl)pyridine	CPMK-alcohol	? mM NADPH	—	Wild-type control with organic solvent (in s^-1 * mM^-1) \| Control is not mutant value in aqueous phase, but WT value in organic solvent
S237G	Activity - Michaelis-Menten	Km (mM), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm), with the assay with 5% Ethanol as control	Ethanol	10%	—	3.13	2.15	mM	100 mM sodium phosphate buffer	6	30°C	2-(4-Chlorobenzoyl)pyridine	CPMK-alcohol	? mM NADPH	—	Wild-type control with organic solvent (in mM) \| Control is not mutant value in aqueous phase, but WT value in organic solvent
S237G	Activity - Michaelis-Menten	kcat (1/s), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm), with the assay with 5% Ethanol as control	Ethanol	10%	—	20.82	72.05	1/s	100 mM sodium phosphate buffer	6	30°C	2-(4-Chlorobenzoyl)pyridine	CPMK-alcohol	? mM NADPH	—	Wild-type control with organic solvent (in 1/s) \| Control is not mutant value in aqueous phase, but WT value in organic solvent
S237G	Activity - Michaelis-Menten	kcat/Km (s^-1 mM^-1), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm), with the assay with 5% Ethanol as control	Ethanol	10%	—	6.65	33.51	s^-1 * mM^-1	100 mM sodium phosphate buffer	6	30°C	2-(4-Chlorobenzoyl)pyridine	CPMK-alcohol	? mM NADPH	—	Wild-type control with organic solvent (in s^-1 * mM^-1) \| Control is not mutant value in aqueous phase, but WT value in organic solvent
V231D	Activity - Michaelis-Menten	Km (mM), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm), with the assay with 5% Ethanol as control	Ethanol	5%	—	24.5	3.62	mM	100 mM sodium phosphate buffer	6	30°C	Methyl 1-Boc-4-hydroxypiperidine-2-carboxylate	Methyl N-Boc-4-oxopiperidine-2-carboxylate	? mM NADPH	—	Wild-type control with organic solvent (in mM) \| Control is not mutant value in aqueous phase, but WT value in organic solvent
V231D	Activity - Michaelis-Menten	kcat (1/s), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm), with the assay with 5% Ethanol as control	Ethanol	5%	—	33.78	1.71	1/s	100 mM sodium phosphate buffer	6	30°C	Methyl 1-Boc-4-hydroxypiperidine-2-carboxylate	Methyl N-Boc-4-oxopiperidine-2-carboxylate	? mM NADPH	—	Wild-type control with organic solvent (in 1/s) \| Control is not mutant value in aqueous phase, but WT value in organic solvent
V231D	Activity - Michaelis-Menten	kcat/Km (s^-1 mM^-1), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm), with the assay with 5% Ethanol as control	Ethanol	5%	—	1.38	0.47	s^-1 * mM^-1	100 mM sodium phosphate buffer	6	30°C	Methyl 1-Boc-4-hydroxypiperidine-2-carboxylate	Methyl N-Boc-4-oxopiperidine-2-carboxylate	? mM NADPH	—	Wild-type control with organic solvent (in s^-1 * mM^-1) \| Control is not mutant value in aqueous phase, but WT value in organic solvent
S237G	Activity - Michaelis-Menten	Km (mM), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm), with the assay with 5% Ethanol as control	Ethanol	5%	—	24.5	14.7	mM	100 mM sodium phosphate buffer	6	30°C	Methyl 1-Boc-4-hydroxypiperidine-2-carboxylate	Methyl N-Boc-4-oxopiperidine-2-carboxylate	? mM NADPH	—	Wild-type control with organic solvent (in mM) \| Control is not mutant value in aqueous phase, but WT value in organic solvent
S237G	Activity - Michaelis-Menten	kcat (1/s), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm), with the assay with 5% Ethanol as control	Ethanol	5%	—	33.78	265.79	1/s	100 mM sodium phosphate buffer	6	30°C	Methyl 1-Boc-4-hydroxypiperidine-2-carboxylate	Methyl N-Boc-4-oxopiperidine-2-carboxylate	? mM NADPH	—	Wild-type control with organic solvent (in 1/s) \| Control is not mutant value in aqueous phase, but WT value in organic solvent
S237G	Activity - Michaelis-Menten	kcat/Km (s^-1 mM^-1), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm), with the assay with 5% Ethanol as control	Ethanol	5%	—	1.38	18.08	s^-1 * mM^-1	100 mM sodium phosphate buffer	6	30°C	Methyl 1-Boc-4-hydroxypiperidine-2-carboxylate	Methyl N-Boc-4-oxopiperidine-2-carboxylate	? mM NADPH	—	Wild-type control with organic solvent (in s^-1 * mM^-1) \| Control is not mutant value in aqueous phase, but WT value in organic solvent

WT	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	%	100 mM sodium phosphate buffer	6	30°C	? mM 2-(4-Chlorobenzoyl)pyridine	CPMK-alcohol	? mM NADPH	—	Classical aqueous control (in %) \| Reference measured with 5% ethanol
WT	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	%	100 mM sodium phosphate buffer	6	30°C	? mM 2-(4-Chlorobenzoyl)pyridine	CPMK-alcohol	? mM NADPH	—	Classical aqueous control (in %) \| Reference measured with 5% ethanol
WT	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	%	100 mM sodium phosphate buffer	6	30°C	? mM 2-(4-Chlorobenzoyl)pyridine	CPMK-alcohol	? mM NADPH	—	Classical aqueous control (in %) \| Reference measured with 5% ethanol
WT	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	%	100 mM sodium phosphate buffer	6	30°C	? mM 2-(4-Chlorobenzoyl)pyridine	CPMK-alcohol	? mM NADPH	—	Classical aqueous control (in %) \| Reference measured with 5% ethanol
WT	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	%	100 mM sodium phosphate buffer	6	30°C	? mM 2-(4-Chlorobenzoyl)pyridine	CPMK-alcohol	? mM NADPH	—	Classical aqueous control (in %) \| Reference measured with 5% ethanol
WT	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	%	100 mM sodium phosphate buffer	6	30°C	? mM 2-(4-Chlorobenzoyl)pyridine	CPMK-alcohol	? mM NADPH	—	Classical aqueous control (in %) \| Reference measured with 5% ethanol
WT	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	%	100 mM sodium phosphate buffer	6	30°C	? mM 2-(4-Chlorobenzoyl)pyridine	CPMK-alcohol	? mM NADPH	—	Classical aqueous control (in %) \| Reference measured with 5% ethanol
WT	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	%	100 mM sodium phosphate buffer	6	30°C	? mM 2-(4-Chlorobenzoyl)pyridine	CPMK-alcohol	? mM NADPH	—	Classical aqueous control (in %) \| Reference measured with 5% ethanol
WT	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	%	100 mM sodium phosphate buffer	6	30°C	? mM 2-(4-Chlorobenzoyl)pyridine	CPMK-alcohol	? mM NADPH	—	Classical aqueous control (in %) \| Reference measured with 5% ethanol
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	—	23.0	%	100 mM sodium phosphate buffer	6	30°C	? mM 2-(4-Chlorobenzoyl)pyridine	CPMK-alcohol	? mM NADPH	—	Classical aqueous control (in %) \| Reference measured with 5% ethanol
WT	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	%	100 mM sodium phosphate buffer	6	30°C	? mM 2-(4-Chlorobenzoyl)pyridine	CPMK-alcohol	? mM NADPH	—	Classical aqueous control (in %) \| Reference measured with 5% ethanol
WT	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	%	100 mM sodium phosphate buffer	6	30°C	? mM 2-(4-Chlorobenzoyl)pyridine	CPMK-alcohol	? mM NADPH	—	Classical aqueous control (in %) \| Reference measured with 5% ethanol
WT	Activity - Classical	Activity measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm), with the assay with 5% Ethanol as control	Dimethyl Sulfoxide (DMSO)	1%	100.0	—	71.1	%	100 mM sodium phosphate buffer	6	30°C	? mM 2-(4-Chlorobenzoyl)pyridine	CPMK-alcohol	? mM NADPH	—	Classical aqueous control (in %) \| Reference measured with 5% ethanol
WT	Activity - Classical	Activity measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm), with the assay with 5% Ethanol as control	Dimethyl Sulfoxide (DMSO)	2%	100.0	—	52.5	%	100 mM sodium phosphate buffer	6	30°C	? mM 2-(4-Chlorobenzoyl)pyridine	CPMK-alcohol	? mM NADPH	—	Classical aqueous control (in %) \| Reference measured with 5% ethanol
WT	Activity - Classical	Activity measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm), with the assay with 5% Ethanol as control	Dimethyl Sulfoxide (DMSO)	3%	100.0	—	44.4	%	100 mM sodium phosphate buffer	6	30°C	? mM 2-(4-Chlorobenzoyl)pyridine	CPMK-alcohol	? mM NADPH	—	Classical aqueous control (in %) \| Reference measured with 5% ethanol
WT	Activity - Classical	Activity measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm), with the assay with 5% Ethanol as control	Dimethyl Sulfoxide (DMSO)	5%	100.0	—	25.9	%	100 mM sodium phosphate buffer	6	30°C	? mM 2-(4-Chlorobenzoyl)pyridine	CPMK-alcohol	? mM NADPH	—	Classical aqueous control (in %) \| Reference measured with 5% ethanol
WT	Activity - Classical	Activity measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm), with the assay with 5% Ethanol as control	Dimethyl Sulfoxide (DMSO)	7.5%	100.0	—	16.1	%	100 mM sodium phosphate buffer	6	30°C	? mM 2-(4-Chlorobenzoyl)pyridine	CPMK-alcohol	? mM NADPH	—	Classical aqueous control (in %) \| Reference measured with 5% ethanol
WT	Activity - Classical	Activity measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm), with the assay with 5% Ethanol as control	Dimethyl Sulfoxide (DMSO)	10%	100.0	—	15.7	%	100 mM sodium phosphate buffer	6	30°C	? mM 2-(4-Chlorobenzoyl)pyridine	CPMK-alcohol	? mM NADPH	—	Classical aqueous control (in %) \| Reference measured with 5% ethanol
WT	Activity - Classical	Activity measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm), with the assay with 5% Ethanol as control	1,4-Dioxane	1.9%	100.0	—	100.0	%	100 mM sodium phosphate buffer	6	30°C	? mM 2-(4-Chlorobenzoyl)pyridine	CPMK-alcohol	? mM NADPH	—	Classical aqueous control (in %) \| Reference measured with 5% ethanol
WT	Activity - Classical	Activity measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm), with the assay with 5% Ethanol as control	1,4-Dioxane	3.7%	100.0	—	84.6	%	100 mM sodium phosphate buffer	6	30°C	? mM 2-(4-Chlorobenzoyl)pyridine	CPMK-alcohol	? mM NADPH	—	Classical aqueous control (in %) \| Reference measured with 5% ethanol
WT	Activity - Classical	Activity measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm), with the assay with 5% Ethanol as control	1,4-Dioxane	5.7%	100.0	—	74.7	%	100 mM sodium phosphate buffer	6	30°C	? mM 2-(4-Chlorobenzoyl)pyridine	CPMK-alcohol	? mM NADPH	—	Classical aqueous control (in %) \| Reference measured with 5% ethanol
WT	Activity - Classical	Activity measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm), with the assay with 5% Ethanol as control	1,4-Dioxane	7.5%	100.0	—	72.5	%	100 mM sodium phosphate buffer	6	30°C	? mM 2-(4-Chlorobenzoyl)pyridine	CPMK-alcohol	? mM NADPH	—	Classical aqueous control (in %) \| Reference measured with 5% ethanol
WT	Activity - Classical	Activity measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm), with the assay with 5% Ethanol as control	1,4-Dioxane	9.3%	100.0	—	35.7	%	100 mM sodium phosphate buffer	6	30°C	? mM 2-(4-Chlorobenzoyl)pyridine	CPMK-alcohol	? mM NADPH	—	Classical aqueous control (in %) \| Reference measured with 5% ethanol
WT	Activity - Classical	Activity measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm), with the assay with 5% Ethanol as control	1,4-Dioxane	11.25%	100.0	—	28.6	%	100 mM sodium phosphate buffer	6	30°C	? mM 2-(4-Chlorobenzoyl)pyridine	CPMK-alcohol	? mM NADPH	—	Classical aqueous control (in %) \| Reference measured with 5% ethanol
WT	Activity - Classical	Activity measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm), with the assay with 5% Ethanol as control	1,4-Dioxane	15%	100.0	—	17.0	%	100 mM sodium phosphate buffer	6	30°C	? mM 2-(4-Chlorobenzoyl)pyridine	CPMK-alcohol	? mM NADPH	—	Classical aqueous control (in %) \| Reference measured with 5% ethanol
WT	Activity - Classical	Activity measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm), with the assay with 5% Ethanol as control	Dimethylformamide (DMF)	1%	100.0	—	91.8	%	100 mM sodium phosphate buffer	6	30°C	? mM 2-(4-Chlorobenzoyl)pyridine	CPMK-alcohol	? mM NADPH	—	Classical aqueous control (in %) \| Reference measured with 5% ethanol
WT	Activity - Classical	Activity measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm), with the assay with 5% Ethanol as control	Dimethylformamide (DMF)	2.5%	100.0	—	40.4	%	100 mM sodium phosphate buffer	6	30°C	? mM 2-(4-Chlorobenzoyl)pyridine	CPMK-alcohol	? mM NADPH	—	Classical aqueous control (in %) \| Reference measured with 5% ethanol
WT	Activity - Classical	Activity measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm), with the assay with 5% Ethanol as control	Dimethylformamide (DMF)	5%	100.0	—	24.6	%	100 mM sodium phosphate buffer	6	30°C	? mM 2-(4-Chlorobenzoyl)pyridine	CPMK-alcohol	? mM NADPH	—	Classical aqueous control (in %) \| Reference measured with 5% ethanol
WT	Activity - Classical	Activity measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm), with the assay with 5% Ethanol as control	Dimethylformamide (DMF)	7.5%	100.0	—	10.5	%	100 mM sodium phosphate buffer	6	30°C	? mM 2-(4-Chlorobenzoyl)pyridine	CPMK-alcohol	? mM NADPH	—	Classical aqueous control (in %) \| Reference measured with 5% ethanol
WT	Activity - Classical	Activity measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm), with the assay with 5% Ethanol as control	2,2,2-Trifluoroethanol (TFE)	1%	100.0	—	87.2	%	100 mM sodium phosphate buffer	6	30°C	? mM 2-(4-Chlorobenzoyl)pyridine	CPMK-alcohol	? mM NADPH	—	Classical aqueous control (in %) \| Reference measured with 5% ethanol
WT	Activity - Classical	Activity measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm), with the assay with 5% Ethanol as control	2,2,2-Trifluoroethanol (TFE)	2%	100.0	—	90.6	%	100 mM sodium phosphate buffer	6	30°C	? mM 2-(4-Chlorobenzoyl)pyridine	CPMK-alcohol	? mM NADPH	—	Classical aqueous control (in %) \| Reference measured with 5% ethanol
WT	Activity - Classical	Activity measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm), with the assay with 5% Ethanol as control	2,2,2-Trifluoroethanol (TFE)	5%	100.0	—	62.6	%	100 mM sodium phosphate buffer	6	30°C	? mM 2-(4-Chlorobenzoyl)pyridine	CPMK-alcohol	? mM NADPH	—	Classical aqueous control (in %) \| Reference measured with 5% ethanol
WT	Activity - Classical	Activity measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm), with the assay with 5% Ethanol as control	2,2,2-Trifluoroethanol (TFE)	7.5%	100.0	—	46.2	%	100 mM sodium phosphate buffer	6	30°C	? mM 2-(4-Chlorobenzoyl)pyridine	CPMK-alcohol	? mM NADPH	—	Classical aqueous control (in %) \| Reference measured with 5% ethanol
WT	Activity - Classical	Activity measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm), with the assay with 5% Ethanol as control	2,2,2-Trifluoroethanol (TFE)	10%	100.0	—	31.0	%	100 mM sodium phosphate buffer	6	30°C	? mM 2-(4-Chlorobenzoyl)pyridine	CPMK-alcohol	? mM NADPH	—	Classical aqueous control (in %) \| Reference measured with 5% ethanol
WT	Activity - Classical	Activity measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm), with the assay with 5% Ethanol as control	2,2,2-Trifluoroethanol (TFE)	15%	100.0	—	11.0	%	100 mM sodium phosphate buffer	6	30°C	? mM 2-(4-Chlorobenzoyl)pyridine	CPMK-alcohol	? mM NADPH	—	Classical aqueous control (in %) \| Reference measured with 5% ethanol
WT	Activity - Classical	Activity measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm), with the assay with 5% Ethanol as control	Ethanol	2.5%	100.0	—	52.0	%	100 mM sodium phosphate buffer	6	30°C	? mM Methyl 1-Boc-4-hydroxypiperidine-2-carboxylate	Methyl N-Boc-4-oxopiperidine-2-carboxylate	? mM NADPH	—	Classical aqueous control (in %) \| Reference measured with 5% ethanol
WT	Activity - Classical	Activity measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm), with the assay with 5% Ethanol as control	Ethanol	5%	100.0	—	32.0	%	100 mM sodium phosphate buffer	6	30°C	? mM Methyl 1-Boc-4-hydroxypiperidine-2-carboxylate	Methyl N-Boc-4-oxopiperidine-2-carboxylate	? mM NADPH	—	Classical aqueous control (in %) \| Reference measured with 5% ethanol
WT	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	—	27.4	%	100 mM sodium phosphate buffer	6	30°C	? mM Methyl 1-Boc-4-hydroxypiperidine-2-carboxylate	Methyl N-Boc-4-oxopiperidine-2-carboxylate	? mM NADPH	—	Classical aqueous control (in %) \| Reference measured with 5% ethanol
WT	Activity - Classical	Activity measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm), with the assay with 5% Ethanol as control	Ethanol	10%	100.0	—	21.1	%	100 mM sodium phosphate buffer	6	30°C	? mM Methyl 1-Boc-4-hydroxypiperidine-2-carboxylate	Methyl N-Boc-4-oxopiperidine-2-carboxylate	? mM NADPH	—	Classical aqueous control (in %) \| Reference measured with 5% ethanol
WT	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	—	14.3	%	100 mM sodium phosphate buffer	6	30°C	? mM Methyl 1-Boc-4-hydroxypiperidine-2-carboxylate	Methyl N-Boc-4-oxopiperidine-2-carboxylate	? mM NADPH	—	Classical aqueous control (in %) \| Reference measured with 5% ethanol
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	—	7.4	%	100 mM sodium phosphate buffer	6	30°C	? mM Methyl 1-Boc-4-hydroxypiperidine-2-carboxylate	Methyl N-Boc-4-oxopiperidine-2-carboxylate	? mM NADPH	—	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	30°C	? mM 2-(4-Chlorobenzoyl)pyridine	CPMK-alcohol	? mM NADPH	—	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	30°C	? mM 2-(4-Chlorobenzoyl)pyridine	CPMK-alcohol	? mM NADPH	—	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	30°C	? mM 2-(4-Chlorobenzoyl)pyridine	CPMK-alcohol	? mM NADPH	—	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	30°C	? mM 2-(4-Chlorobenzoyl)pyridine	CPMK-alcohol	? mM NADPH	—	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	30°C	? mM 2-(4-Chlorobenzoyl)pyridine	CPMK-alcohol	? mM NADPH	—	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	30°C	? mM 2-(4-Chlorobenzoyl)pyridine	CPMK-alcohol	? mM NADPH	—	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	30°C	? mM 2-(4-Chlorobenzoyl)pyridine	CPMK-alcohol	? mM NADPH	—	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	30°C	? mM 2-(4-Chlorobenzoyl)pyridine	CPMK-alcohol	? mM NADPH	—	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	30°C	? mM 2-(4-Chlorobenzoyl)pyridine	CPMK-alcohol	? mM NADPH	—	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	30°C	? mM 2-(4-Chlorobenzoyl)pyridine	CPMK-alcohol	? mM NADPH	—	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	30°C	? mM 2-(4-Chlorobenzoyl)pyridine	CPMK-alcohol	? mM NADPH	—	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	30°C	? mM 2-(4-Chlorobenzoyl)pyridine	CPMK-alcohol	? mM NADPH	—	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	30°C	? mM 2-(4-Chlorobenzoyl)pyridine	CPMK-alcohol	? mM NADPH	—	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	30°C	? mM 2-(4-Chlorobenzoyl)pyridine	CPMK-alcohol	? mM NADPH	—	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	30°C	? mM 2-(4-Chlorobenzoyl)pyridine	CPMK-alcohol	? mM NADPH	—	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	30°C	? mM 2-(4-Chlorobenzoyl)pyridine	CPMK-alcohol	? mM NADPH	—	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	30°C	? mM 2-(4-Chlorobenzoyl)pyridine	CPMK-alcohol	? mM NADPH	—	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	30°C	? mM 2-(4-Chlorobenzoyl)pyridine	CPMK-alcohol	? mM NADPH	—	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	30°C	? mM 2-(4-Chlorobenzoyl)pyridine	CPMK-alcohol	? mM NADPH	—	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	10%	100.0	49.4	58.5	%	100 mM sodium phosphate buffer	6	30°C	? mM 2-(4-Chlorobenzoyl)pyridine	CPMK-alcohol	? mM NADPH	—	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	12.5%	100.0	33.1	38.2	%	100 mM sodium phosphate buffer	6	30°C	? mM 2-(4-Chlorobenzoyl)pyridine	CPMK-alcohol	? mM NADPH	—	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	15%	100.0	23.0	33.2	%	100 mM sodium phosphate buffer	6	30°C	? mM 2-(4-Chlorobenzoyl)pyridine	CPMK-alcohol	? mM NADPH	—	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	17.5%	100.0	19.7	19.7	%	100 mM sodium phosphate buffer	6	30°C	? mM 2-(4-Chlorobenzoyl)pyridine	CPMK-alcohol	? mM NADPH	—	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	20%	100.0	13.0	16.4	%	100 mM sodium phosphate buffer	6	30°C	? mM 2-(4-Chlorobenzoyl)pyridine	CPMK-alcohol	? mM NADPH	—	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	Dimethyl Sulfoxide (DMSO)	1%	100.0	71.1	87.5	%	100 mM sodium phosphate buffer	6	30°C	? mM 2-(4-Chlorobenzoyl)pyridine	CPMK-alcohol	? mM NADPH	—	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	Dimethyl Sulfoxide (DMSO)	2%	100.0	52.5	85.9	%	100 mM sodium phosphate buffer	6	30°C	? mM 2-(4-Chlorobenzoyl)pyridine	CPMK-alcohol	? mM NADPH	—	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	Dimethyl Sulfoxide (DMSO)	3%	100.0	44.4	67.9	%	100 mM sodium phosphate buffer	6	30°C	? mM 2-(4-Chlorobenzoyl)pyridine	CPMK-alcohol	? mM NADPH	—	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	Dimethyl Sulfoxide (DMSO)	5%	100.0	25.9	60.8	%	100 mM sodium phosphate buffer	6	30°C	? mM 2-(4-Chlorobenzoyl)pyridine	CPMK-alcohol	? mM NADPH	—	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	Dimethyl Sulfoxide (DMSO)	7.5%	100.0	16.1	35.2	%	100 mM sodium phosphate buffer	6	30°C	? mM 2-(4-Chlorobenzoyl)pyridine	CPMK-alcohol	? mM NADPH	—	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	Dimethyl Sulfoxide (DMSO)	10%	100.0	15.7	29.3	%	100 mM sodium phosphate buffer	6	30°C	? mM 2-(4-Chlorobenzoyl)pyridine	CPMK-alcohol	? mM NADPH	—	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	Dimethyl Sulfoxide (DMSO)	1%	100.0	71.1	80.4	%	100 mM sodium phosphate buffer	6	30°C	? mM 2-(4-Chlorobenzoyl)pyridine	CPMK-alcohol	? mM NADPH	—	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	Dimethyl Sulfoxide (DMSO)	2%	100.0	52.5	71.1	%	100 mM sodium phosphate buffer	6	30°C	? mM 2-(4-Chlorobenzoyl)pyridine	CPMK-alcohol	? mM NADPH	—	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	Dimethyl Sulfoxide (DMSO)	3%	100.0	44.4	59.7	%	100 mM sodium phosphate buffer	6	30°C	? mM 2-(4-Chlorobenzoyl)pyridine	CPMK-alcohol	? mM NADPH	—	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	Dimethyl Sulfoxide (DMSO)	5%	100.0	25.9	42.8	%	100 mM sodium phosphate buffer	6	30°C	? mM 2-(4-Chlorobenzoyl)pyridine	CPMK-alcohol	? mM NADPH	—	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	Dimethyl Sulfoxide (DMSO)	7.5%	100.0	16.1	25.9	%	100 mM sodium phosphate buffer	6	30°C	? mM 2-(4-Chlorobenzoyl)pyridine	CPMK-alcohol	? mM NADPH	—	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	Dimethyl Sulfoxide (DMSO)	10%	100.0	15.7	20.6	%	100 mM sodium phosphate buffer	6	30°C	? mM 2-(4-Chlorobenzoyl)pyridine	CPMK-alcohol	? mM NADPH	—	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	1,4-Dioxane	1.9%	100.0	100.0	71.4	%	100 mM sodium phosphate buffer	6	30°C	? mM 2-(4-Chlorobenzoyl)pyridine	CPMK-alcohol	? mM NADPH	—	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	1,4-Dioxane	3.7%	100.0	84.6	42.3	%	100 mM sodium phosphate buffer	6	30°C	? mM 2-(4-Chlorobenzoyl)pyridine	CPMK-alcohol	? mM NADPH	—	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	1,4-Dioxane	5.7%	100.0	74.7	28.0	%	100 mM sodium phosphate buffer	6	30°C	? mM 2-(4-Chlorobenzoyl)pyridine	CPMK-alcohol	? mM NADPH	—	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	1,4-Dioxane	7.5%	100.0	74.6	14.8	%	100 mM sodium phosphate buffer	6	30°C	? mM 2-(4-Chlorobenzoyl)pyridine	CPMK-alcohol	? mM NADPH	—	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	1,4-Dioxane	9.3%	100.0	35.7	6.5	%	100 mM sodium phosphate buffer	6	30°C	? mM 2-(4-Chlorobenzoyl)pyridine	CPMK-alcohol	? mM NADPH	—	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	1,4-Dioxane	11.25%	100.0	28.6	0.0	%	100 mM sodium phosphate buffer	6	30°C	? mM 2-(4-Chlorobenzoyl)pyridine	CPMK-alcohol	? mM NADPH	—	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	1,4-Dioxane	1.9%	100.0	100.0	87.9	%	100 mM sodium phosphate buffer	6	30°C	? mM 2-(4-Chlorobenzoyl)pyridine	CPMK-alcohol	? mM NADPH	—	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	1,4-Dioxane	3.7%	100.0	84.6	58.2	%	100 mM sodium phosphate buffer	6	30°C	? mM 2-(4-Chlorobenzoyl)pyridine	CPMK-alcohol	? mM NADPH	—	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	1,4-Dioxane	5.7%	100.0	74.7	45.0	%	100 mM sodium phosphate buffer	6	30°C	? mM 2-(4-Chlorobenzoyl)pyridine	CPMK-alcohol	? mM NADPH	—	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	1,4-Dioxane	7.5%	100.0	74.6	39.6	%	100 mM sodium phosphate buffer	6	30°C	? mM 2-(4-Chlorobenzoyl)pyridine	CPMK-alcohol	? mM NADPH	—	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	1,4-Dioxane	9.3%	100.0	35.7	16.5	%	100 mM sodium phosphate buffer	6	30°C	? mM 2-(4-Chlorobenzoyl)pyridine	CPMK-alcohol	? mM NADPH	—	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	1,4-Dioxane	11.25%	100.0	28.6	12.6	%	100 mM sodium phosphate buffer	6	30°C	? mM 2-(4-Chlorobenzoyl)pyridine	CPMK-alcohol	? mM NADPH	—	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	1,4-Dioxane	15%	100.0	28.5	7.2	%	100 mM sodium phosphate buffer	6	30°C	? mM 2-(4-Chlorobenzoyl)pyridine	CPMK-alcohol	? mM NADPH	—	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	Dimethylformamide (DMF)	1%	100.0	91.6	71.4	%	100 mM sodium phosphate buffer	6	30°C	? mM 2-(4-Chlorobenzoyl)pyridine	CPMK-alcohol	? mM NADPH	—	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	Dimethylformamide (DMF)	2.5%	100.0	40.4	52.0	%	100 mM sodium phosphate buffer	6	30°C	? mM 2-(4-Chlorobenzoyl)pyridine	CPMK-alcohol	? mM NADPH	—	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	Dimethylformamide (DMF)	5%	100.0	24.6	26.3	%	100 mM sodium phosphate buffer	6	30°C	? mM 2-(4-Chlorobenzoyl)pyridine	CPMK-alcohol	? mM NADPH	—	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	Dimethylformamide (DMF)	7.5%	100.0	10.5	14.6	%	100 mM sodium phosphate buffer	6	30°C	? mM 2-(4-Chlorobenzoyl)pyridine	CPMK-alcohol	? mM NADPH	—	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	Dimethylformamide (DMF)	1%	100.0	91.6	115.8	%	100 mM sodium phosphate buffer	6	30°C	? mM 2-(4-Chlorobenzoyl)pyridine	CPMK-alcohol	? mM NADPH	—	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	Dimethylformamide (DMF)	2.5%	100.0	40.4	35.7	%	100 mM sodium phosphate buffer	6	30°C	? mM 2-(4-Chlorobenzoyl)pyridine	CPMK-alcohol	? mM NADPH	—	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	Dimethylformamide (DMF)	5%	100.0	24.6	20.5	%	100 mM sodium phosphate buffer	6	30°C	? mM 2-(4-Chlorobenzoyl)pyridine	CPMK-alcohol	? mM NADPH	—	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	Dimethylformamide (DMF)	7.5%	100.0	10.5	10.0	%	100 mM sodium phosphate buffer	6	30°C	? mM 2-(4-Chlorobenzoyl)pyridine	CPMK-alcohol	? mM NADPH	—	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	2,2,2-Trifluoroethanol (TFE)	1%	100.0	87.2	76.1	%	100 mM sodium phosphate buffer	6	30°C	? mM 2-(4-Chlorobenzoyl)pyridine	CPMK-alcohol	? mM NADPH	—	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	2,2,2-Trifluoroethanol (TFE)	2%	100.0	90.6	67.9	%	100 mM sodium phosphate buffer	6	30°C	? mM 2-(4-Chlorobenzoyl)pyridine	CPMK-alcohol	? mM NADPH	—	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	2,2,2-Trifluoroethanol (TFE)	5%	100.0	62.6	40.5	%	100 mM sodium phosphate buffer	6	30°C	? mM 2-(4-Chlorobenzoyl)pyridine	CPMK-alcohol	? mM NADPH	—	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	2,2,2-Trifluoroethanol (TFE)	7.5%	100.0	46.2	14.8	%	100 mM sodium phosphate buffer	6	30°C	? mM 2-(4-Chlorobenzoyl)pyridine	CPMK-alcohol	? mM NADPH	—	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	2,2,2-Trifluoroethanol (TFE)	10%	100.0	31.0	4.3	%	100 mM sodium phosphate buffer	6	30°C	? mM 2-(4-Chlorobenzoyl)pyridine	CPMK-alcohol	? mM NADPH	—	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	2,2,2-Trifluoroethanol (TFE)	1%	100.0	87.2	102.3	%	100 mM sodium phosphate buffer	6	30°C	? mM 2-(4-Chlorobenzoyl)pyridine	CPMK-alcohol	? mM NADPH	—	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	2,2,2-Trifluoroethanol (TFE)	2%	100.0	90.6	108.1	%	100 mM sodium phosphate buffer	6	30°C	? mM 2-(4-Chlorobenzoyl)pyridine	CPMK-alcohol	? mM NADPH	—	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	2,2,2-Trifluoroethanol (TFE)	5%	100.0	62.6	62.0	%	100 mM sodium phosphate buffer	6	30°C	? mM 2-(4-Chlorobenzoyl)pyridine	CPMK-alcohol	? mM NADPH	—	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	2,2,2-Trifluoroethanol (TFE)	7.5%	100.0	46.2	41.6	%	100 mM sodium phosphate buffer	6	30°C	? mM 2-(4-Chlorobenzoyl)pyridine	CPMK-alcohol	? mM NADPH	—	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	2,2,2-Trifluoroethanol (TFE)	10%	100.0	31.0	18.2	%	100 mM sodium phosphate buffer	6	30°C	? mM 2-(4-Chlorobenzoyl)pyridine	CPMK-alcohol	? mM NADPH	—	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	2,2,2-Trifluoroethanol (TFE)	15%	100.0	11.0	5.8	%	100 mM sodium phosphate buffer	6	30°C	? mM 2-(4-Chlorobenzoyl)pyridine	CPMK-alcohol	? mM NADPH	—	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	2.5%	100.0	52.0	98.9	%	100 mM sodium phosphate buffer	6	30°C	? mM Methyl 1-Boc-4-hydroxypiperidine-2-carboxylate	Methyl N-Boc-4-oxopiperidine-2-carboxylate	? mM NADPH	—	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	5%	100.0	32.0	93.1	%	100 mM sodium phosphate buffer	6	30°C	? mM Methyl 1-Boc-4-hydroxypiperidine-2-carboxylate	Methyl N-Boc-4-oxopiperidine-2-carboxylate	? mM NADPH	—	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	27.4	65.7	%	100 mM sodium phosphate buffer	6	30°C	? mM Methyl 1-Boc-4-hydroxypiperidine-2-carboxylate	Methyl N-Boc-4-oxopiperidine-2-carboxylate	? mM NADPH	—	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	21.1	57.1	%	100 mM sodium phosphate buffer	6	30°C	? mM Methyl 1-Boc-4-hydroxypiperidine-2-carboxylate	Methyl N-Boc-4-oxopiperidine-2-carboxylate	? mM NADPH	—	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	14.3	43.4	%	100 mM sodium phosphate buffer	6	30°C	? mM Methyl 1-Boc-4-hydroxypiperidine-2-carboxylate	Methyl N-Boc-4-oxopiperidine-2-carboxylate	? mM NADPH	—	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	7.4	34.3	%	100 mM sodium phosphate buffer	6	30°C	? mM Methyl 1-Boc-4-hydroxypiperidine-2-carboxylate	Methyl N-Boc-4-oxopiperidine-2-carboxylate	? mM NADPH	—	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	2.5%	100.0	52.0	85.7	%	100 mM sodium phosphate buffer	6	30°C	? mM Methyl 1-Boc-4-hydroxypiperidine-2-carboxylate	Methyl N-Boc-4-oxopiperidine-2-carboxylate	? mM NADPH	—	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	5%	100.0	32.0	62.8	%	100 mM sodium phosphate buffer	6	30°C	? mM Methyl 1-Boc-4-hydroxypiperidine-2-carboxylate	Methyl N-Boc-4-oxopiperidine-2-carboxylate	? mM NADPH	—	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	27.4	51.4	%	100 mM sodium phosphate buffer	6	30°C	? mM Methyl 1-Boc-4-hydroxypiperidine-2-carboxylate	Methyl N-Boc-4-oxopiperidine-2-carboxylate	? mM NADPH	—	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	10%	100.0	21.1	42.3	%	100 mM sodium phosphate buffer	6	30°C	? mM Methyl 1-Boc-4-hydroxypiperidine-2-carboxylate	Methyl N-Boc-4-oxopiperidine-2-carboxylate	? mM NADPH	—	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	12.5%	100.0	14.3	28.6	%	100 mM sodium phosphate buffer	6	30°C	? mM Methyl 1-Boc-4-hydroxypiperidine-2-carboxylate	Methyl N-Boc-4-oxopiperidine-2-carboxylate	? mM NADPH	—	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	15%	100.0	7.4	22.3	%	100 mM sodium phosphate buffer	6	30°C	? mM Methyl 1-Boc-4-hydroxypiperidine-2-carboxylate	Methyl N-Boc-4-oxopiperidine-2-carboxylate	? mM NADPH	—	Classical aqueous control (in %) \| Reference measured with 5% ethanol
V231D	Activity - Michaelis-Menten	Km (mM), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm), with the assay with 5% Ethanol as control	Ethanol	5%	—	2.29	22.74	mM	100 mM sodium phosphate buffer	6	30°C	2-(4-Chlorobenzoyl)pyridine	CPMK-alcohol	? mM NADPH	—	Wild-type control with organic solvent (in mM) \| Control is not mutant value in aqueous phase, but WT value in organic solvent
V231D	Activity - Michaelis-Menten	kcat (1/s), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm), with the assay with 5% Ethanol as control	Ethanol	5%	—	32.05	12.81	1/s	100 mM sodium phosphate buffer	6	30°C	2-(4-Chlorobenzoyl)pyridine	CPMK-alcohol	? mM NADPH	—	Wild-type control with organic solvent (in 1/s) \| Control is not mutant value in aqueous phase, but WT value in organic solvent
V231D	Activity - Michaelis-Menten	kcat/Km (s^-1 mM^-1), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm), with the assay with 5% Ethanol as control	Ethanol	5%	—	14.0	0.56	s^-1 * mM^-1	100 mM sodium phosphate buffer	6	30°C	2-(4-Chlorobenzoyl)pyridine	CPMK-alcohol	? mM NADPH	—	Wild-type control with organic solvent (in s^-1 * mM^-1) \| Control is not mutant value in aqueous phase, but WT value in organic solvent
S237G	Activity - Michaelis-Menten	Km (mM), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm), with the assay with 5% Ethanol as control	Ethanol	5%	—	2.29	1.07	mM	100 mM sodium phosphate buffer	6	30°C	2-(4-Chlorobenzoyl)pyridine	CPMK-alcohol	? mM NADPH	—	Wild-type control with organic solvent (in mM) \| Control is not mutant value in aqueous phase, but WT value in organic solvent
S237G	Activity - Michaelis-Menten	kcat (1/s), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm), with the assay with 5% Ethanol as control	Ethanol	5%	—	32.05	97.7	1/s	100 mM sodium phosphate buffer	6	30°C	2-(4-Chlorobenzoyl)pyridine	CPMK-alcohol	? mM NADPH	—	Wild-type control with organic solvent (in 1/s) \| Control is not mutant value in aqueous phase, but WT value in organic solvent
S237G	Activity - Michaelis-Menten	kcat/Km (s^-1 mM^-1), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm), with the assay with 5% Ethanol as control	Ethanol	5%	—	14.0	91.56	s^-1 * mM^-1	100 mM sodium phosphate buffer	6	30°C	2-(4-Chlorobenzoyl)pyridine	CPMK-alcohol	? mM NADPH	—	Wild-type control with organic solvent (in s^-1 * mM^-1) \| Control is not mutant value in aqueous phase, but WT value in organic solvent
V231D	Activity - Michaelis-Menten	Km (mM), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm), with the assay with 5% Ethanol as control	Ethanol	10%	—	3.13	44.23	mM	100 mM sodium phosphate buffer	6	30°C	2-(4-Chlorobenzoyl)pyridine	CPMK-alcohol	? mM NADPH	—	Wild-type control with organic solvent (in mM) \| Control is not mutant value in aqueous phase, but WT value in organic solvent
V231D	Activity - Michaelis-Menten	kcat (1/s), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm), with the assay with 5% Ethanol as control	Ethanol	10%	—	20.82	8.68	1/s	100 mM sodium phosphate buffer	6	30°C	2-(4-Chlorobenzoyl)pyridine	CPMK-alcohol	? mM NADPH	—	Wild-type control with organic solvent (in 1/s) \| Control is not mutant value in aqueous phase, but WT value in organic solvent
V231D	Activity - Michaelis-Menten	kcat/Km (s^-1 mM^-1), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm), with the assay with 5% Ethanol as control	Ethanol	10%	—	6.65	0.196	s^-1 * mM^-1	100 mM sodium phosphate buffer	6	30°C	2-(4-Chlorobenzoyl)pyridine	CPMK-alcohol	? mM NADPH	—	Wild-type control with organic solvent (in s^-1 * mM^-1) \| Control is not mutant value in aqueous phase, but WT value in organic solvent
S237G	Activity - Michaelis-Menten	Km (mM), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm), with the assay with 5% Ethanol as control	Ethanol	10%	—	3.13	2.15	mM	100 mM sodium phosphate buffer	6	30°C	2-(4-Chlorobenzoyl)pyridine	CPMK-alcohol	? mM NADPH	—	Wild-type control with organic solvent (in mM) \| Control is not mutant value in aqueous phase, but WT value in organic solvent
S237G	Activity - Michaelis-Menten	kcat (1/s), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm), with the assay with 5% Ethanol as control	Ethanol	10%	—	20.82	72.05	1/s	100 mM sodium phosphate buffer	6	30°C	2-(4-Chlorobenzoyl)pyridine	CPMK-alcohol	? mM NADPH	—	Wild-type control with organic solvent (in 1/s) \| Control is not mutant value in aqueous phase, but WT value in organic solvent
S237G	Activity - Michaelis-Menten	kcat/Km (s^-1 mM^-1), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm), with the assay with 5% Ethanol as control	Ethanol	10%	—	6.65	33.51	s^-1 * mM^-1	100 mM sodium phosphate buffer	6	30°C	2-(4-Chlorobenzoyl)pyridine	CPMK-alcohol	? mM NADPH	—	Wild-type control with organic solvent (in s^-1 * mM^-1) \| Control is not mutant value in aqueous phase, but WT value in organic solvent
V231D	Activity - Michaelis-Menten	Km (mM), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm), with the assay with 5% Ethanol as control	Ethanol	5%	—	24.5	3.62	mM	100 mM sodium phosphate buffer	6	30°C	Methyl 1-Boc-4-hydroxypiperidine-2-carboxylate	Methyl N-Boc-4-oxopiperidine-2-carboxylate	? mM NADPH	—	Wild-type control with organic solvent (in mM) \| Control is not mutant value in aqueous phase, but WT value in organic solvent
V231D	Activity - Michaelis-Menten	kcat (1/s), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm), with the assay with 5% Ethanol as control	Ethanol	5%	—	33.78	1.71	1/s	100 mM sodium phosphate buffer	6	30°C	Methyl 1-Boc-4-hydroxypiperidine-2-carboxylate	Methyl N-Boc-4-oxopiperidine-2-carboxylate	? mM NADPH	—	Wild-type control with organic solvent (in 1/s) \| Control is not mutant value in aqueous phase, but WT value in organic solvent
V231D	Activity - Michaelis-Menten	kcat/Km (s^-1 mM^-1), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm), with the assay with 5% Ethanol as control	Ethanol	5%	—	1.38	0.47	s^-1 * mM^-1	100 mM sodium phosphate buffer	6	30°C	Methyl 1-Boc-4-hydroxypiperidine-2-carboxylate	Methyl N-Boc-4-oxopiperidine-2-carboxylate	? mM NADPH	—	Wild-type control with organic solvent (in s^-1 * mM^-1) \| Control is not mutant value in aqueous phase, but WT value in organic solvent
S237G	Activity - Michaelis-Menten	Km (mM), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm), with the assay with 5% Ethanol as control	Ethanol	5%	—	24.5	14.7	mM	100 mM sodium phosphate buffer	6	30°C	Methyl 1-Boc-4-hydroxypiperidine-2-carboxylate	Methyl N-Boc-4-oxopiperidine-2-carboxylate	? mM NADPH	—	Wild-type control with organic solvent (in mM) \| Control is not mutant value in aqueous phase, but WT value in organic solvent
S237G	Activity - Michaelis-Menten	kcat (1/s), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm), with the assay with 5% Ethanol as control	Ethanol	5%	—	33.78	265.79	1/s	100 mM sodium phosphate buffer	6	30°C	Methyl 1-Boc-4-hydroxypiperidine-2-carboxylate	Methyl N-Boc-4-oxopiperidine-2-carboxylate	? mM NADPH	—	Wild-type control with organic solvent (in 1/s) \| Control is not mutant value in aqueous phase, but WT value in organic solvent
S237G	Activity - Michaelis-Menten	kcat/Km (s^-1 mM^-1), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm), with the assay with 5% Ethanol as control	Ethanol	5%	—	1.38	18.08	s^-1 * mM^-1	100 mM sodium phosphate buffer	6	30°C	Methyl 1-Boc-4-hydroxypiperidine-2-carboxylate	Methyl N-Boc-4-oxopiperidine-2-carboxylate	? mM NADPH	—	Wild-type control with organic solvent (in s^-1 * mM^-1) \| Control is not mutant value in aqueous phase, but WT value in organic solvent

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Mutations in this dataset (3)

WT V231D S237G

Visualization : Activity — Classical

One bar per measurement. Colour = solvent, shade = solvent volume. — — — 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 — Michaelis-Menten

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

Mutation Effect — 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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