Haloalkane dehalogenase DhaA from Rhodococcus rhodochrous

Enzyme Description

Species

Extremophile

EC Number

3.8.1.5 ↗

Sequence

Length: 293 amino acids

MSEIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSSYLWRNIIPHVAPSHRCIAPDLIGMGKSDKPDLDYFFDDHVRYLDAFIEALGLEEVVLVIHDWGSALGFHWAKRNPERVKGIACMEFIRPIPTWDEWPEFARETFQAFRTADVGRELIIDQNAFIEGALPKCVVRPLTEVEMDHYREPFLKPVDREPLWRFPNELPIAGEPANIVALVEAYMNWLHQSPVPKLLFWGTPGVLIPPAEAARLAESLPNCKTVDIGPGLHYLQEDNPDLIGSEIARWLPAL

Veronika Stepankova et al. (2013) — Organic co-solvents affect activity, stability and enantioselectivity of haloalkane dehalogenases

Biotechnology Journal · doi:10.1002/biot.201200378 ↗ · Activity - Classical Stability - C50 Stability - Tm

136 measurements ▼

Database ID

UniProt: P0A3G2 ↗

Sequence Annotation

Inferred - from protein name

Protein Source

Recombinant, host bacterium Escherichia coli BL21

Experimental Data (136 measurements)

136 measurements — page 7 of 7

Property	Assay	Solvent	Solvent Volume	Aqueous Reference	Measured Value	Units	Solution	pH	Temperature	Substrate(s)	Product(s)	Cofactor(s)	Shaking	Comments
Stability - C50	Volume fraction of organic solvent at which 50% of enzyme activity is lost, measured by absorbance spectrophotometry (colorimetric assay, iodine, mercuric thiocyanate and ferric ammonium sulfate reaction product Fe–SCN absorbance measurement, 460 nm) in the presence of organic solvent	1,4-Dioxane	—	—	4	% (v/v)	glycine 100 mM	8.6	37	1-Iodohexane	1-Hexanol , Iodide ion	—	—	Not applicable control (in % (v/v))
Stability - C50	Volume fraction of organic solvent at which 50% of enzyme activity is lost, measured by absorbance spectrophotometry (colorimetric assay, iodine, mercuric thiocyanate and ferric ammonium sulfate reaction product Fe–SCN absorbance measurement, 460 nm) in the presence of organic solvent	Acetone	—	—	16	% (v/v)	glycine 100 mM	8.6	37	1-Iodohexane	1-Hexanol , Iodide ion	—	—	Not applicable control (in % (v/v))
Stability - C50	Volume fraction of organic solvent at which 50% of enzyme activity is lost, measured by absorbance spectrophotometry (colorimetric assay, iodine, mercuric thiocyanate and ferric ammonium sulfate reaction product Fe–SCN absorbance measurement, 460 nm) in the presence of organic solvent	Isopropanol	—	—	9	% (v/v)	glycine 100 mM	8.6	37	1-Iodohexane	1-Hexanol , Iodide ion	—	—	Not applicable control (in % (v/v))
Stability - C50	Volume fraction of organic solvent at which 50% of enzyme activity is lost, measured by absorbance spectrophotometry (colorimetric assay, iodine, mercuric thiocyanate and ferric ammonium sulfate reaction product Fe–SCN absorbance measurement, 460 nm) in the presence of organic solvent	Tetrahydrofuran (THF)	—	—	7	% (v/v)	glycine 100 mM	8.6	37	1-Iodohexane	1-Hexanol , Iodide ion	—	—	Not applicable control (in % (v/v))
Stability - Tm	Tm measured by circular dichroism in the presence of organic solvent	Ethylene Glycol	20%	51	50	°C	50 mM phosphate buffer	7.5	—	—	—	—	—	Classical aqueous control (in °C), averaged between circular dichroism and DSC measurements
Stability - Tm	Tm measured by DSC in the presence of organic solvent	Dimethylformamide (DMF)	20%	51	34	°C	50 mM phosphate buffer	7.5	—	—	—	—	—	Classical aqueous control (in °C), averaged between circular dichroism and DSC measurements
Stability - Tm	Tm measured by DSC in the presence of organic solvent	Dimethyl Sulfoxide (DMSO)	20%	51	47	°C	50 mM phosphate buffer	7.5	—	—	—	—	—	Classical aqueous control (in °C), averaged between circular dichroism and DSC measurements
Stability - Tm	Tm measured by circular dichroism in the presence of organic solvent	Methanol	20%	51	40	°C	50 mM phosphate buffer	7.5	—	—	—	—	—	Classical aqueous control (in °C), averaged between circular dichroism and DSC measurements
Stability - Tm	Tm measured by circular dichroism in the presence of organic solvent	1,4-Dioxane	20%	51	30	°C	50 mM phosphate buffer	7.5	—	—	—	—	—	Classical aqueous control (in °C), averaged between circular dichroism and DSC measurements
Stability - Tm	Tm measured by circular dichroism in the presence of organic solvent	Acetonitrile	10%	51	37	°C	50 mM phosphate buffer	7.5	—	—	—	—	—	Classical aqueous control (in °C), averaged between circular dichroism and DSC measurements
Stability - Tm	Tm measured by circular dichroism in the presence of organic solvent	Acetonitrile	20%	51	<10	°C	50 mM phosphate buffer	7.5	—	—	—	—	—	Classical aqueous control (in °C), averaged between circular dichroism and DSC measurements
Stability - Tm	Tm measured by circular dichroism in the presence of organic solvent	Ethanol	20%	51	35	°C	50 mM phosphate buffer	7.5	—	—	—	—	—	Classical aqueous control (in °C), averaged between circular dichroism and DSC measurements
Stability - Tm	Tm measured by circular dichroism in the presence of organic solvent	Acetone	20%	51	31	°C	50 mM phosphate buffer	7.5	—	—	—	—	—	Classical aqueous control (in °C), averaged between circular dichroism and DSC measurements
Stability - Tm	Tm measured by circular dichroism in the presence of organic solvent	Isopropanol	20%	51	33	°C	50 mM phosphate buffer	7.5	—	—	—	—	—	Classical aqueous control (in °C), averaged between circular dichroism and DSC measurements
Stability - Tm	Tm measured by circular dichroism in the presence of organic solvent	Tetrahydrofuran (THF)	10%	51	32	°C	50 mM phosphate buffer	7.5	—	—	—	—	—	Classical aqueous control (in °C), averaged between circular dichroism and DSC measurements
Stability - Tm	Tm measured by circular dichroism in the presence of organic solvent	Tetrahydrofuran (THF)	20%	51	0	°C	50 mM phosphate buffer	7.5	—	—	—	—	—	Classical aqueous control (in °C), averaged between circular dichroism and DSC measurements

—	Activity - Classical	Activity measured by absorbance spectrophotometry (colorimetric assay, iodine, mercuric thiocyanate and ferric ammonium sulfate reaction product Fe–SCN absorbance measurement, 460 nm) in the presence of organic solvent	Glycerol	5%	100	112	%	glycine 100 mM	8.6	37	? mM 1-Iodohexane	1-Hexanol, Iodide ion	—	—	Classical aqueous control (in %)
—	Activity - Classical	Activity measured by absorbance spectrophotometry (colorimetric assay, iodine, mercuric thiocyanate and ferric ammonium sulfate reaction product Fe–SCN absorbance measurement, 460 nm) in the presence of organic solvent	Formamide	5%	100	150	%	glycine 100 mM	8.6	37	? mM 1-Iodohexane	1-Hexanol, Iodide ion	—	—	Classical aqueous control (in %)
—	Activity - Classical	Activity measured by absorbance spectrophotometry (colorimetric assay, iodine, mercuric thiocyanate and ferric ammonium sulfate reaction product Fe–SCN absorbance measurement, 460 nm) in the presence of organic solvent	Ethylene Glycol	5%	100	113	%	glycine 100 mM	8.6	37	? mM 1-Iodohexane	1-Hexanol, Iodide ion	—	—	Classical aqueous control (in %)
—	Activity - Classical	Activity measured by absorbance spectrophotometry (colorimetric assay, iodine, mercuric thiocyanate and ferric ammonium sulfate reaction product Fe–SCN absorbance measurement, 460 nm) in the presence of organic solvent	PEG-1000	5%	100	95	%	glycine 100 mM	8.6	37	? mM 1-Iodohexane	1-Hexanol, Iodide ion	—	—	Classical aqueous control (in %)
—	Activity - Classical	Activity measured by absorbance spectrophotometry (colorimetric assay, iodine, mercuric thiocyanate and ferric ammonium sulfate reaction product Fe–SCN absorbance measurement, 460 nm) in the presence of organic solvent	PEG-10000	5%	100	94	%	glycine 100 mM	8.6	37	? mM 1-Iodohexane	1-Hexanol, Iodide ion	—	—	Classical aqueous control (in %)
—	Activity - Classical	Activity measured by absorbance spectrophotometry (colorimetric assay, iodine, mercuric thiocyanate and ferric ammonium sulfate reaction product Fe–SCN absorbance measurement, 460 nm) in the presence of organic solvent	Dimethylformamide (DMF)	5%	100	97	%	glycine 100 mM	8.6	37	? mM 1-Iodohexane	1-Hexanol, Iodide ion	—	—	Classical aqueous control (in %)
—	Activity - Classical	Activity measured by absorbance spectrophotometry (colorimetric assay, iodine, mercuric thiocyanate and ferric ammonium sulfate reaction product Fe–SCN absorbance measurement, 460 nm) in the presence of organic solvent	Dimethyl Sulfoxide (DMSO)	5%	100	111	%	glycine 100 mM	8.6	37	? mM 1-Iodohexane	1-Hexanol, Iodide ion	—	—	Classical aqueous control (in %)
—	Activity - Classical	Activity measured by absorbance spectrophotometry (colorimetric assay, iodine, mercuric thiocyanate and ferric ammonium sulfate reaction product Fe–SCN absorbance measurement, 460 nm) in the presence of organic solvent	Methanol	5%	100	150	%	glycine 100 mM	8.6	37	? mM 1-Iodohexane	1-Hexanol, Iodide ion	—	—	Classical aqueous control (in %)
—	Activity - Classical	Activity measured by absorbance spectrophotometry (colorimetric assay, iodine, mercuric thiocyanate and ferric ammonium sulfate reaction product Fe–SCN absorbance measurement, 460 nm) in the presence of organic solvent	1,4-Dioxane	5%	100	33	%	glycine 100 mM	8.6	37	? mM 1-Iodohexane	1-Hexanol, Iodide ion	—	—	Classical aqueous control (in %)
—	Activity - Classical	Activity measured by absorbance spectrophotometry (colorimetric assay, iodine, mercuric thiocyanate and ferric ammonium sulfate reaction product Fe–SCN absorbance measurement, 460 nm) in the presence of organic solvent	Acetonitrile	5%	100	100	%	glycine 100 mM	8.6	37	? mM 1-Iodohexane	1-Hexanol, Iodide ion	—	—	Classical aqueous control (in %)
—	Activity - Classical	Activity measured by absorbance spectrophotometry (colorimetric assay, iodine, mercuric thiocyanate and ferric ammonium sulfate reaction product Fe–SCN absorbance measurement, 460 nm) in the presence of organic solvent	Ethanol	5%	100	120	%	glycine 100 mM	8.6	37	? mM 1-Iodohexane	1-Hexanol, Iodide ion	—	—	Classical aqueous control (in %)
—	Activity - Classical	Activity measured by absorbance spectrophotometry (colorimetric assay, iodine, mercuric thiocyanate and ferric ammonium sulfate reaction product Fe–SCN absorbance measurement, 460 nm) in the presence of organic solvent	Acetone	5%	100	108	%	glycine 100 mM	8.6	37	? mM 1-Iodohexane	1-Hexanol, Iodide ion	—	—	Classical aqueous control (in %)
—	Activity - Classical	Activity measured by absorbance spectrophotometry (colorimetric assay, iodine, mercuric thiocyanate and ferric ammonium sulfate reaction product Fe–SCN absorbance measurement, 460 nm) in the presence of organic solvent	Isopropanol	5%	100	111	%	glycine 100 mM	8.6	37	? mM 1-Iodohexane	1-Hexanol, Iodide ion	—	—	Classical aqueous control (in %)
—	Activity - Classical	Activity measured by absorbance spectrophotometry (colorimetric assay, iodine, mercuric thiocyanate and ferric ammonium sulfate reaction product Fe–SCN absorbance measurement, 460 nm) in the presence of organic solvent	Tetrahydrofuran (THF)	5%	100	75	%	glycine 100 mM	8.6	37	? mM 1-Iodohexane	1-Hexanol, Iodide ion	—	—	Classical aqueous control (in %)
—	Activity - Classical	Activity measured by absorbance spectrophotometry (colorimetric assay, iodine, mercuric thiocyanate and ferric ammonium sulfate reaction product Fe–SCN absorbance measurement, 460 nm) in the presence of organic solvent	Glycerol	10%	100	120	%	glycine 100 mM	8.6	37	? mM 1-Iodohexane	1-Hexanol, Iodide ion	—	—	Classical aqueous control (in %)
—	Activity - Classical	Activity measured by absorbance spectrophotometry (colorimetric assay, iodine, mercuric thiocyanate and ferric ammonium sulfate reaction product Fe–SCN absorbance measurement, 460 nm) in the presence of organic solvent	Formamide	10%	100	112	%	glycine 100 mM	8.6	37	? mM 1-Iodohexane	1-Hexanol, Iodide ion	—	—	Classical aqueous control (in %)
—	Activity - Classical	Activity measured by absorbance spectrophotometry (colorimetric assay, iodine, mercuric thiocyanate and ferric ammonium sulfate reaction product Fe–SCN absorbance measurement, 460 nm) in the presence of organic solvent	Ethylene Glycol	10%	100	110	%	glycine 100 mM	8.6	37	? mM 1-Iodohexane	1-Hexanol, Iodide ion	—	—	Classical aqueous control (in %)
—	Activity - Classical	Activity measured by absorbance spectrophotometry (colorimetric assay, iodine, mercuric thiocyanate and ferric ammonium sulfate reaction product Fe–SCN absorbance measurement, 460 nm) in the presence of organic solvent	PEG-1000	10%	100	100	%	glycine 100 mM	8.6	37	? mM 1-Iodohexane	1-Hexanol, Iodide ion	—	—	Classical aqueous control (in %)
—	Activity - Classical	Activity measured by absorbance spectrophotometry (colorimetric assay, iodine, mercuric thiocyanate and ferric ammonium sulfate reaction product Fe–SCN absorbance measurement, 460 nm) in the presence of organic solvent	PEG-10000	10%	100	96	%	glycine 100 mM	8.6	37	? mM 1-Iodohexane	1-Hexanol, Iodide ion	—	—	Classical aqueous control (in %)
—	Activity - Classical	Activity measured by absorbance spectrophotometry (colorimetric assay, iodine, mercuric thiocyanate and ferric ammonium sulfate reaction product Fe–SCN absorbance measurement, 460 nm) in the presence of organic solvent	Dimethylformamide (DMF)	10%	100	97	%	glycine 100 mM	8.6	37	? mM 1-Iodohexane	1-Hexanol, Iodide ion	—	—	Classical aqueous control (in %)
—	Activity - Classical	Activity measured by absorbance spectrophotometry (colorimetric assay, iodine, mercuric thiocyanate and ferric ammonium sulfate reaction product Fe–SCN absorbance measurement, 460 nm) in the presence of organic solvent	Dimethyl Sulfoxide (DMSO)	10%	100	113	%	glycine 100 mM	8.6	37	? mM 1-Iodohexane	1-Hexanol, Iodide ion	—	—	Classical aqueous control (in %)
—	Activity - Classical	Activity measured by absorbance spectrophotometry (colorimetric assay, iodine, mercuric thiocyanate and ferric ammonium sulfate reaction product Fe–SCN absorbance measurement, 460 nm) in the presence of organic solvent	Methanol	10%	100	141	%	glycine 100 mM	8.6	37	? mM 1-Iodohexane	1-Hexanol, Iodide ion	—	—	Classical aqueous control (in %)
—	Activity - Classical	Activity measured by absorbance spectrophotometry (colorimetric assay, iodine, mercuric thiocyanate and ferric ammonium sulfate reaction product Fe–SCN absorbance measurement, 460 nm) in the presence of organic solvent	1,4-Dioxane	10%	100	29	%	glycine 100 mM	8.6	37	? mM 1-Iodohexane	1-Hexanol, Iodide ion	—	—	Classical aqueous control (in %)
—	Activity - Classical	Activity measured by absorbance spectrophotometry (colorimetric assay, iodine, mercuric thiocyanate and ferric ammonium sulfate reaction product Fe–SCN absorbance measurement, 460 nm) in the presence of organic solvent	Acetonitrile	10%	100	32	%	glycine 100 mM	8.6	37	? mM 1-Iodohexane	1-Hexanol, Iodide ion	—	—	Classical aqueous control (in %)
—	Activity - Classical	Activity measured by absorbance spectrophotometry (colorimetric assay, iodine, mercuric thiocyanate and ferric ammonium sulfate reaction product Fe–SCN absorbance measurement, 460 nm) in the presence of organic solvent	Ethanol	10%	100	117	%	glycine 100 mM	8.6	37	? mM 1-Iodohexane	1-Hexanol, Iodide ion	—	—	Classical aqueous control (in %)
—	Activity - Classical	Activity measured by absorbance spectrophotometry (colorimetric assay, iodine, mercuric thiocyanate and ferric ammonium sulfate reaction product Fe–SCN absorbance measurement, 460 nm) in the presence of organic solvent	Acetone	10%	100	89	%	glycine 100 mM	8.6	37	? mM 1-Iodohexane	1-Hexanol, Iodide ion	—	—	Classical aqueous control (in %)
—	Activity - Classical	Activity measured by absorbance spectrophotometry (colorimetric assay, iodine, mercuric thiocyanate and ferric ammonium sulfate reaction product Fe–SCN absorbance measurement, 460 nm) in the presence of organic solvent	Isopropanol	10%	100	36	%	glycine 100 mM	8.6	37	? mM 1-Iodohexane	1-Hexanol, Iodide ion	—	—	Classical aqueous control (in %)
—	Activity - Classical	Activity measured by absorbance spectrophotometry (colorimetric assay, iodine, mercuric thiocyanate and ferric ammonium sulfate reaction product Fe–SCN absorbance measurement, 460 nm) in the presence of organic solvent	Tetrahydrofuran (THF)	10%	100	13	%	glycine 100 mM	8.6	37	? mM 1-Iodohexane	1-Hexanol, Iodide ion	—	—	Classical aqueous control (in %)
—	Activity - Classical	Activity measured by absorbance spectrophotometry (colorimetric assay, iodine, mercuric thiocyanate and ferric ammonium sulfate reaction product Fe–SCN absorbance measurement, 460 nm) in the presence of organic solvent	Glycerol	20%	100	125	%	glycine 100 mM	8.6	37	? mM 1-Iodohexane	1-Hexanol, Iodide ion	—	—	Classical aqueous control (in %)
—	Activity - Classical	Activity measured by absorbance spectrophotometry (colorimetric assay, iodine, mercuric thiocyanate and ferric ammonium sulfate reaction product Fe–SCN absorbance measurement, 460 nm) in the presence of organic solvent	Formamide	20%	100	62	%	glycine 100 mM	8.6	37	? mM 1-Iodohexane	1-Hexanol, Iodide ion	—	—	Classical aqueous control (in %)
—	Activity - Classical	Activity measured by absorbance spectrophotometry (colorimetric assay, iodine, mercuric thiocyanate and ferric ammonium sulfate reaction product Fe–SCN absorbance measurement, 460 nm) in the presence of organic solvent	Ethylene Glycol	20%	100	111	%	glycine 100 mM	8.6	37	? mM 1-Iodohexane	1-Hexanol, Iodide ion	—	—	Classical aqueous control (in %)
—	Activity - Classical	Activity measured by absorbance spectrophotometry (colorimetric assay, iodine, mercuric thiocyanate and ferric ammonium sulfate reaction product Fe–SCN absorbance measurement, 460 nm) in the presence of organic solvent	PEG-1000	20%	100	100	%	glycine 100 mM	8.6	37	? mM 1-Iodohexane	1-Hexanol, Iodide ion	—	—	Classical aqueous control (in %)
—	Activity - Classical	Activity measured by absorbance spectrophotometry (colorimetric assay, iodine, mercuric thiocyanate and ferric ammonium sulfate reaction product Fe–SCN absorbance measurement, 460 nm) in the presence of organic solvent	PEG-10000	20%	100	73	%	glycine 100 mM	8.6	37	? mM 1-Iodohexane	1-Hexanol, Iodide ion	—	—	Classical aqueous control (in %)
—	Activity - Classical	Activity measured by absorbance spectrophotometry (colorimetric assay, iodine, mercuric thiocyanate and ferric ammonium sulfate reaction product Fe–SCN absorbance measurement, 460 nm) in the presence of organic solvent	Dimethylformamide (DMF)	20%	100	46	%	glycine 100 mM	8.6	37	? mM 1-Iodohexane	1-Hexanol, Iodide ion	—	—	Classical aqueous control (in %)
—	Activity - Classical	Activity measured by absorbance spectrophotometry (colorimetric assay, iodine, mercuric thiocyanate and ferric ammonium sulfate reaction product Fe–SCN absorbance measurement, 460 nm) in the presence of organic solvent	Dimethyl Sulfoxide (DMSO)	20%	100	107	%	glycine 100 mM	8.6	37	? mM 1-Iodohexane	1-Hexanol, Iodide ion	—	—	Classical aqueous control (in %)
—	Activity - Classical	Activity measured by absorbance spectrophotometry (colorimetric assay, iodine, mercuric thiocyanate and ferric ammonium sulfate reaction product Fe–SCN absorbance measurement, 460 nm) in the presence of organic solvent	Methanol	20%	100	76	%	glycine 100 mM	8.6	37	? mM 1-Iodohexane	1-Hexanol, Iodide ion	—	—	Classical aqueous control (in %)
—	Activity - Classical	Activity measured by absorbance spectrophotometry (colorimetric assay, iodine, mercuric thiocyanate and ferric ammonium sulfate reaction product Fe–SCN absorbance measurement, 460 nm) in the presence of organic solvent	1,4-Dioxane	20%	100	0	%	glycine 100 mM	8.6	37	? mM 1-Iodohexane	1-Hexanol, Iodide ion	—	—	Classical aqueous control (in %)
—	Activity - Classical	Activity measured by absorbance spectrophotometry (colorimetric assay, iodine, mercuric thiocyanate and ferric ammonium sulfate reaction product Fe–SCN absorbance measurement, 460 nm) in the presence of organic solvent	Acetonitrile	20%	100	0	%	glycine 100 mM	8.6	37	? mM 1-Iodohexane	1-Hexanol, Iodide ion	—	—	Classical aqueous control (in %)
—	Activity - Classical	Activity measured by absorbance spectrophotometry (colorimetric assay, iodine, mercuric thiocyanate and ferric ammonium sulfate reaction product Fe–SCN absorbance measurement, 460 nm) in the presence of organic solvent	Ethanol	20%	100	56	%	glycine 100 mM	8.6	37	? mM 1-Iodohexane	1-Hexanol, Iodide ion	—	—	Classical aqueous control (in %)
—	Activity - Classical	Activity measured by absorbance spectrophotometry (colorimetric assay, iodine, mercuric thiocyanate and ferric ammonium sulfate reaction product Fe–SCN absorbance measurement, 460 nm) in the presence of organic solvent	Acetone	20%	100	11	%	glycine 100 mM	8.6	37	? mM 1-Iodohexane	1-Hexanol, Iodide ion	—	—	Classical aqueous control (in %)
—	Activity - Classical	Activity measured by absorbance spectrophotometry (colorimetric assay, iodine, mercuric thiocyanate and ferric ammonium sulfate reaction product Fe–SCN absorbance measurement, 460 nm) in the presence of organic solvent	Isopropanol	20%	100	0	%	glycine 100 mM	8.6	37	? mM 1-Iodohexane	1-Hexanol, Iodide ion	—	—	Classical aqueous control (in %)
—	Activity - Classical	Activity measured by absorbance spectrophotometry (colorimetric assay, iodine, mercuric thiocyanate and ferric ammonium sulfate reaction product Fe–SCN absorbance measurement, 460 nm) in the presence of organic solvent	Tetrahydrofuran (THF)	20%	100	0	%	glycine 100 mM	8.6	37	? mM 1-Iodohexane	1-Hexanol, Iodide ion	—	—	Classical aqueous control (in %)
—	Activity - Classical	Activity measured by absorbance spectrophotometry (colorimetric assay, iodine, mercuric thiocyanate and ferric ammonium sulfate reaction product Fe–SCN absorbance measurement, 460 nm) in the presence of organic solvent	Glycerol	30%	100	107	%	glycine 100 mM	8.6	37	? mM 1-Iodohexane	1-Hexanol, Iodide ion	—	—	Classical aqueous control (in %)
—	Activity - Classical	Activity measured by absorbance spectrophotometry (colorimetric assay, iodine, mercuric thiocyanate and ferric ammonium sulfate reaction product Fe–SCN absorbance measurement, 460 nm) in the presence of organic solvent	Formamide	30%	100	50	%	glycine 100 mM	8.6	37	? mM 1-Iodohexane	1-Hexanol, Iodide ion	—	—	Classical aqueous control (in %)
—	Activity - Classical	Activity measured by absorbance spectrophotometry (colorimetric assay, iodine, mercuric thiocyanate and ferric ammonium sulfate reaction product Fe–SCN absorbance measurement, 460 nm) in the presence of organic solvent	Ethylene Glycol	30%	100	121	%	glycine 100 mM	8.6	37	? mM 1-Iodohexane	1-Hexanol, Iodide ion	—	—	Classical aqueous control (in %)
—	Activity - Classical	Activity measured by absorbance spectrophotometry (colorimetric assay, iodine, mercuric thiocyanate and ferric ammonium sulfate reaction product Fe–SCN absorbance measurement, 460 nm) in the presence of organic solvent	PEG-1000	30%	100	108	%	glycine 100 mM	8.6	37	? mM 1-Iodohexane	1-Hexanol, Iodide ion	—	—	Classical aqueous control (in %)
—	Activity - Classical	Activity measured by absorbance spectrophotometry (colorimetric assay, iodine, mercuric thiocyanate and ferric ammonium sulfate reaction product Fe–SCN absorbance measurement, 460 nm) in the presence of organic solvent	PEG-10000	30%	100	67	%	glycine 100 mM	8.6	37	? mM 1-Iodohexane	1-Hexanol, Iodide ion	—	—	Classical aqueous control (in %)
—	Activity - Classical	Activity measured by absorbance spectrophotometry (colorimetric assay, iodine, mercuric thiocyanate and ferric ammonium sulfate reaction product Fe–SCN absorbance measurement, 460 nm) in the presence of organic solvent	Dimethylformamide (DMF)	30%	100	17	%	glycine 100 mM	8.6	37	? mM 1-Iodohexane	1-Hexanol, Iodide ion	—	—	Classical aqueous control (in %)
—	Activity - Classical	Activity measured by absorbance spectrophotometry (colorimetric assay, iodine, mercuric thiocyanate and ferric ammonium sulfate reaction product Fe–SCN absorbance measurement, 460 nm) in the presence of organic solvent	Dimethyl Sulfoxide (DMSO)	30%	100	95	%	glycine 100 mM	8.6	37	? mM 1-Iodohexane	1-Hexanol, Iodide ion	—	—	Classical aqueous control (in %)
—	Activity - Classical	Activity measured by absorbance spectrophotometry (colorimetric assay, iodine, mercuric thiocyanate and ferric ammonium sulfate reaction product Fe–SCN absorbance measurement, 460 nm) in the presence of organic solvent	Methanol	30%	100	23	%	glycine 100 mM	8.6	37	? mM 1-Iodohexane	1-Hexanol, Iodide ion	—	—	Classical aqueous control (in %)
—	Activity - Classical	Activity measured by absorbance spectrophotometry (colorimetric assay, iodine, mercuric thiocyanate and ferric ammonium sulfate reaction product Fe–SCN absorbance measurement, 460 nm) in the presence of organic solvent	1,4-Dioxane	30%	100	0	%	glycine 100 mM	8.6	37	? mM 1-Iodohexane	1-Hexanol, Iodide ion	—	—	Classical aqueous control (in %)
—	Activity - Classical	Activity measured by absorbance spectrophotometry (colorimetric assay, iodine, mercuric thiocyanate and ferric ammonium sulfate reaction product Fe–SCN absorbance measurement, 460 nm) in the presence of organic solvent	Acetonitrile	30%	100	0	%	glycine 100 mM	8.6	37	? mM 1-Iodohexane	1-Hexanol, Iodide ion	—	—	Classical aqueous control (in %)
—	Activity - Classical	Activity measured by absorbance spectrophotometry (colorimetric assay, iodine, mercuric thiocyanate and ferric ammonium sulfate reaction product Fe–SCN absorbance measurement, 460 nm) in the presence of organic solvent	Ethanol	30%	100	7	%	glycine 100 mM	8.6	37	? mM 1-Iodohexane	1-Hexanol, Iodide ion	—	—	Classical aqueous control (in %)
—	Activity - Classical	Activity measured by absorbance spectrophotometry (colorimetric assay, iodine, mercuric thiocyanate and ferric ammonium sulfate reaction product Fe–SCN absorbance measurement, 460 nm) in the presence of organic solvent	Acetone	30%	100	0	%	glycine 100 mM	8.6	37	? mM 1-Iodohexane	1-Hexanol, Iodide ion	—	—	Classical aqueous control (in %)
—	Activity - Classical	Activity measured by absorbance spectrophotometry (colorimetric assay, iodine, mercuric thiocyanate and ferric ammonium sulfate reaction product Fe–SCN absorbance measurement, 460 nm) in the presence of organic solvent	Isopropanol	30%	100	0	%	glycine 100 mM	8.6	37	? mM 1-Iodohexane	1-Hexanol, Iodide ion	—	—	Classical aqueous control (in %)
—	Activity - Classical	Activity measured by absorbance spectrophotometry (colorimetric assay, iodine, mercuric thiocyanate and ferric ammonium sulfate reaction product Fe–SCN absorbance measurement, 460 nm) in the presence of organic solvent	Tetrahydrofuran (THF)	30%	100	0	%	glycine 100 mM	8.6	37	? mM 1-Iodohexane	1-Hexanol, Iodide ion	—	—	Classical aqueous control (in %)
—	Activity - Classical	Activity measured by absorbance spectrophotometry (colorimetric assay, iodine, mercuric thiocyanate and ferric ammonium sulfate reaction product Fe–SCN absorbance measurement, 460 nm) in the presence of organic solvent	Glycerol	40%	100	110	%	glycine 100 mM	8.6	37	? mM 1-Iodohexane	1-Hexanol, Iodide ion	—	—	Classical aqueous control (in %)
—	Activity - Classical	Activity measured by absorbance spectrophotometry (colorimetric assay, iodine, mercuric thiocyanate and ferric ammonium sulfate reaction product Fe–SCN absorbance measurement, 460 nm) in the presence of organic solvent	Formamide	40%	100	0	%	glycine 100 mM	8.6	37	? mM 1-Iodohexane	1-Hexanol, Iodide ion	—	—	Classical aqueous control (in %)
—	Activity - Classical	Activity measured by absorbance spectrophotometry (colorimetric assay, iodine, mercuric thiocyanate and ferric ammonium sulfate reaction product Fe–SCN absorbance measurement, 460 nm) in the presence of organic solvent	Ethylene Glycol	40%	100	126	%	glycine 100 mM	8.6	37	? mM 1-Iodohexane	1-Hexanol, Iodide ion	—	—	Classical aqueous control (in %)
—	Activity - Classical	Activity measured by absorbance spectrophotometry (colorimetric assay, iodine, mercuric thiocyanate and ferric ammonium sulfate reaction product Fe–SCN absorbance measurement, 460 nm) in the presence of organic solvent	PEG-1000	40%	100	117	%	glycine 100 mM	8.6	37	? mM 1-Iodohexane	1-Hexanol, Iodide ion	—	—	Classical aqueous control (in %)
—	Activity - Classical	Activity measured by absorbance spectrophotometry (colorimetric assay, iodine, mercuric thiocyanate and ferric ammonium sulfate reaction product Fe–SCN absorbance measurement, 460 nm) in the presence of organic solvent	PEG-10000	40%	100	66	%	glycine 100 mM	8.6	37	? mM 1-Iodohexane	1-Hexanol, Iodide ion	—	—	Classical aqueous control (in %)
—	Activity - Classical	Activity measured by absorbance spectrophotometry (colorimetric assay, iodine, mercuric thiocyanate and ferric ammonium sulfate reaction product Fe–SCN absorbance measurement, 460 nm) in the presence of organic solvent	Dimethylformamide (DMF)	40%	100	0	%	glycine 100 mM	8.6	37	? mM 1-Iodohexane	1-Hexanol, Iodide ion	—	—	Classical aqueous control (in %)
—	Activity - Classical	Activity measured by absorbance spectrophotometry (colorimetric assay, iodine, mercuric thiocyanate and ferric ammonium sulfate reaction product Fe–SCN absorbance measurement, 460 nm) in the presence of organic solvent	Dimethyl Sulfoxide (DMSO)	40%	100	71	%	glycine 100 mM	8.6	37	? mM 1-Iodohexane	1-Hexanol, Iodide ion	—	—	Classical aqueous control (in %)
—	Activity - Classical	Activity measured by absorbance spectrophotometry (colorimetric assay, iodine, mercuric thiocyanate and ferric ammonium sulfate reaction product Fe–SCN absorbance measurement, 460 nm) in the presence of organic solvent	Methanol	40%	100	0	%	glycine 100 mM	8.6	37	? mM 1-Iodohexane	1-Hexanol, Iodide ion	—	—	Classical aqueous control (in %)
—	Activity - Classical	Activity measured by absorbance spectrophotometry (colorimetric assay, iodine, mercuric thiocyanate and ferric ammonium sulfate reaction product Fe–SCN absorbance measurement, 460 nm) in the presence of organic solvent	1,4-Dioxane	40%	100	0	%	glycine 100 mM	8.6	37	? mM 1-Iodohexane	1-Hexanol, Iodide ion	—	—	Classical aqueous control (in %)
—	Activity - Classical	Activity measured by absorbance spectrophotometry (colorimetric assay, iodine, mercuric thiocyanate and ferric ammonium sulfate reaction product Fe–SCN absorbance measurement, 460 nm) in the presence of organic solvent	Acetonitrile	40%	100	0	%	glycine 100 mM	8.6	37	? mM 1-Iodohexane	1-Hexanol, Iodide ion	—	—	Classical aqueous control (in %)
—	Activity - Classical	Activity measured by absorbance spectrophotometry (colorimetric assay, iodine, mercuric thiocyanate and ferric ammonium sulfate reaction product Fe–SCN absorbance measurement, 460 nm) in the presence of organic solvent	Ethanol	40%	100	0	%	glycine 100 mM	8.6	37	? mM 1-Iodohexane	1-Hexanol, Iodide ion	—	—	Classical aqueous control (in %)
—	Activity - Classical	Activity measured by absorbance spectrophotometry (colorimetric assay, iodine, mercuric thiocyanate and ferric ammonium sulfate reaction product Fe–SCN absorbance measurement, 460 nm) in the presence of organic solvent	Acetone	40%	100	0	%	glycine 100 mM	8.6	37	? mM 1-Iodohexane	1-Hexanol, Iodide ion	—	—	Classical aqueous control (in %)
—	Activity - Classical	Activity measured by absorbance spectrophotometry (colorimetric assay, iodine, mercuric thiocyanate and ferric ammonium sulfate reaction product Fe–SCN absorbance measurement, 460 nm) in the presence of organic solvent	Isopropanol	40%	100	0	%	glycine 100 mM	8.6	37	? mM 1-Iodohexane	1-Hexanol, Iodide ion	—	—	Classical aqueous control (in %)
—	Activity - Classical	Activity measured by absorbance spectrophotometry (colorimetric assay, iodine, mercuric thiocyanate and ferric ammonium sulfate reaction product Fe–SCN absorbance measurement, 460 nm) in the presence of organic solvent	Tetrahydrofuran (THF)	40%	100	0	%	glycine 100 mM	8.6	37	? mM 1-Iodohexane	1-Hexanol, Iodide ion	—	—	Classical aqueous control (in %)
—	Activity - Classical	Activity measured by absorbance spectrophotometry (colorimetric assay, iodine, mercuric thiocyanate and ferric ammonium sulfate reaction product Fe–SCN absorbance measurement, 460 nm) in the presence of organic solvent	Glycerol	50%	100	96	%	glycine 100 mM	8.6	37	? mM 1-Iodohexane	1-Hexanol, Iodide ion	—	—	Classical aqueous control (in %)
—	Activity - Classical	Activity measured by absorbance spectrophotometry (colorimetric assay, iodine, mercuric thiocyanate and ferric ammonium sulfate reaction product Fe–SCN absorbance measurement, 460 nm) in the presence of organic solvent	Formamide	50%	100	0	%	glycine 100 mM	8.6	37	? mM 1-Iodohexane	1-Hexanol, Iodide ion	—	—	Classical aqueous control (in %)
—	Activity - Classical	Activity measured by absorbance spectrophotometry (colorimetric assay, iodine, mercuric thiocyanate and ferric ammonium sulfate reaction product Fe–SCN absorbance measurement, 460 nm) in the presence of organic solvent	Ethylene Glycol	50%	100	98	%	glycine 100 mM	8.6	37	? mM 1-Iodohexane	1-Hexanol, Iodide ion	—	—	Classical aqueous control (in %)
—	Activity - Classical	Activity measured by absorbance spectrophotometry (colorimetric assay, iodine, mercuric thiocyanate and ferric ammonium sulfate reaction product Fe–SCN absorbance measurement, 460 nm) in the presence of organic solvent	PEG-1000	50%	100	119	%	glycine 100 mM	8.6	37	? mM 1-Iodohexane	1-Hexanol, Iodide ion	—	—	Classical aqueous control (in %)
—	Activity - Classical	Activity measured by absorbance spectrophotometry (colorimetric assay, iodine, mercuric thiocyanate and ferric ammonium sulfate reaction product Fe–SCN absorbance measurement, 460 nm) in the presence of organic solvent	PEG-10000	50%	100	0	%	glycine 100 mM	8.6	37	? mM 1-Iodohexane	1-Hexanol, Iodide ion	—	—	Classical aqueous control (in %)
—	Activity - Classical	Activity measured by absorbance spectrophotometry (colorimetric assay, iodine, mercuric thiocyanate and ferric ammonium sulfate reaction product Fe–SCN absorbance measurement, 460 nm) in the presence of organic solvent	Dimethylformamide (DMF)	50%	100	0	%	glycine 100 mM	8.6	37	? mM 1-Iodohexane	1-Hexanol, Iodide ion	—	—	Classical aqueous control (in %)
—	Activity - Classical	Activity measured by absorbance spectrophotometry (colorimetric assay, iodine, mercuric thiocyanate and ferric ammonium sulfate reaction product Fe–SCN absorbance measurement, 460 nm) in the presence of organic solvent	Dimethyl Sulfoxide (DMSO)	50%	100	45	%	glycine 100 mM	8.6	37	? mM 1-Iodohexane	1-Hexanol, Iodide ion	—	—	Classical aqueous control (in %)
—	Activity - Classical	Activity measured by absorbance spectrophotometry (colorimetric assay, iodine, mercuric thiocyanate and ferric ammonium sulfate reaction product Fe–SCN absorbance measurement, 460 nm) in the presence of organic solvent	Methanol	50%	100	0	%	glycine 100 mM	8.6	37	? mM 1-Iodohexane	1-Hexanol, Iodide ion	—	—	Classical aqueous control (in %)
—	Activity - Classical	Activity measured by absorbance spectrophotometry (colorimetric assay, iodine, mercuric thiocyanate and ferric ammonium sulfate reaction product Fe–SCN absorbance measurement, 460 nm) in the presence of organic solvent	1,4-Dioxane	50%	100	0	%	glycine 100 mM	8.6	37	? mM 1-Iodohexane	1-Hexanol, Iodide ion	—	—	Classical aqueous control (in %)
—	Activity - Classical	Activity measured by absorbance spectrophotometry (colorimetric assay, iodine, mercuric thiocyanate and ferric ammonium sulfate reaction product Fe–SCN absorbance measurement, 460 nm) in the presence of organic solvent	Acetonitrile	50%	100	0	%	glycine 100 mM	8.6	37	? mM 1-Iodohexane	1-Hexanol, Iodide ion	—	—	Classical aqueous control (in %)
—	Activity - Classical	Activity measured by absorbance spectrophotometry (colorimetric assay, iodine, mercuric thiocyanate and ferric ammonium sulfate reaction product Fe–SCN absorbance measurement, 460 nm) in the presence of organic solvent	Ethanol	50%	100	0	%	glycine 100 mM	8.6	37	? mM 1-Iodohexane	1-Hexanol, Iodide ion	—	—	Classical aqueous control (in %)
—	Activity - Classical	Activity measured by absorbance spectrophotometry (colorimetric assay, iodine, mercuric thiocyanate and ferric ammonium sulfate reaction product Fe–SCN absorbance measurement, 460 nm) in the presence of organic solvent	Acetone	50%	100	0	%	glycine 100 mM	8.6	37	? mM 1-Iodohexane	1-Hexanol, Iodide ion	—	—	Classical aqueous control (in %)
—	Activity - Classical	Activity measured by absorbance spectrophotometry (colorimetric assay, iodine, mercuric thiocyanate and ferric ammonium sulfate reaction product Fe–SCN absorbance measurement, 460 nm) in the presence of organic solvent	Isopropanol	50%	100	0	%	glycine 100 mM	8.6	37	? mM 1-Iodohexane	1-Hexanol, Iodide ion	—	—	Classical aqueous control (in %)
—	Activity - Classical	Activity measured by absorbance spectrophotometry (colorimetric assay, iodine, mercuric thiocyanate and ferric ammonium sulfate reaction product Fe–SCN absorbance measurement, 460 nm) in the presence of organic solvent	Tetrahydrofuran (THF)	50%	100	0	%	glycine 100 mM	8.6	37	? mM 1-Iodohexane	1-Hexanol, Iodide ion	—	—	Classical aqueous control (in %)
—	Activity - Classical	Activity measured by absorbance spectrophotometry (colorimetric assay, iodine, mercuric thiocyanate and ferric ammonium sulfate reaction product Fe–SCN absorbance measurement, 460 nm) in the presence of organic solvent	Glycerol	60%	100	84	%	glycine 100 mM	8.6	37	? mM 1-Iodohexane	1-Hexanol, Iodide ion	—	—	Classical aqueous control (in %)
—	Activity - Classical	Activity measured by absorbance spectrophotometry (colorimetric assay, iodine, mercuric thiocyanate and ferric ammonium sulfate reaction product Fe–SCN absorbance measurement, 460 nm) in the presence of organic solvent	Formamide	60%	100	0	%	glycine 100 mM	8.6	37	? mM 1-Iodohexane	1-Hexanol, Iodide ion	—	—	Classical aqueous control (in %)
—	Activity - Classical	Activity measured by absorbance spectrophotometry (colorimetric assay, iodine, mercuric thiocyanate and ferric ammonium sulfate reaction product Fe–SCN absorbance measurement, 460 nm) in the presence of organic solvent	Ethylene Glycol	60%	100	82	%	glycine 100 mM	8.6	37	? mM 1-Iodohexane	1-Hexanol, Iodide ion	—	—	Classical aqueous control (in %)
—	Activity - Classical	Activity measured by absorbance spectrophotometry (colorimetric assay, iodine, mercuric thiocyanate and ferric ammonium sulfate reaction product Fe–SCN absorbance measurement, 460 nm) in the presence of organic solvent	PEG-1000	60%	100	123	%	glycine 100 mM	8.6	37	? mM 1-Iodohexane	1-Hexanol, Iodide ion	—	—	Classical aqueous control (in %)
—	Activity - Classical	Activity measured by absorbance spectrophotometry (colorimetric assay, iodine, mercuric thiocyanate and ferric ammonium sulfate reaction product Fe–SCN absorbance measurement, 460 nm) in the presence of organic solvent	PEG-10000	60%	100	0	%	glycine 100 mM	8.6	37	? mM 1-Iodohexane	1-Hexanol, Iodide ion	—	—	Classical aqueous control (in %)
—	Activity - Classical	Activity measured by absorbance spectrophotometry (colorimetric assay, iodine, mercuric thiocyanate and ferric ammonium sulfate reaction product Fe–SCN absorbance measurement, 460 nm) in the presence of organic solvent	Dimethylformamide (DMF)	60%	100	0	%	glycine 100 mM	8.6	37	? mM 1-Iodohexane	1-Hexanol, Iodide ion	—	—	Classical aqueous control (in %)
—	Activity - Classical	Activity measured by absorbance spectrophotometry (colorimetric assay, iodine, mercuric thiocyanate and ferric ammonium sulfate reaction product Fe–SCN absorbance measurement, 460 nm) in the presence of organic solvent	Dimethyl Sulfoxide (DMSO)	60%	100	0	%	glycine 100 mM	8.6	37	? mM 1-Iodohexane	1-Hexanol, Iodide ion	—	—	Classical aqueous control (in %)
—	Activity - Classical	Activity measured by absorbance spectrophotometry (colorimetric assay, iodine, mercuric thiocyanate and ferric ammonium sulfate reaction product Fe–SCN absorbance measurement, 460 nm) in the presence of organic solvent	Methanol	60%	100	0	%	glycine 100 mM	8.6	37	? mM 1-Iodohexane	1-Hexanol, Iodide ion	—	—	Classical aqueous control (in %)
—	Activity - Classical	Activity measured by absorbance spectrophotometry (colorimetric assay, iodine, mercuric thiocyanate and ferric ammonium sulfate reaction product Fe–SCN absorbance measurement, 460 nm) in the presence of organic solvent	1,4-Dioxane	60%	100	0	%	glycine 100 mM	8.6	37	? mM 1-Iodohexane	1-Hexanol, Iodide ion	—	—	Classical aqueous control (in %)
—	Activity - Classical	Activity measured by absorbance spectrophotometry (colorimetric assay, iodine, mercuric thiocyanate and ferric ammonium sulfate reaction product Fe–SCN absorbance measurement, 460 nm) in the presence of organic solvent	Acetonitrile	60%	100	0	%	glycine 100 mM	8.6	37	? mM 1-Iodohexane	1-Hexanol, Iodide ion	—	—	Classical aqueous control (in %)
—	Activity - Classical	Activity measured by absorbance spectrophotometry (colorimetric assay, iodine, mercuric thiocyanate and ferric ammonium sulfate reaction product Fe–SCN absorbance measurement, 460 nm) in the presence of organic solvent	Ethanol	60%	100	0	%	glycine 100 mM	8.6	37	? mM 1-Iodohexane	1-Hexanol, Iodide ion	—	—	Classical aqueous control (in %)
—	Activity - Classical	Activity measured by absorbance spectrophotometry (colorimetric assay, iodine, mercuric thiocyanate and ferric ammonium sulfate reaction product Fe–SCN absorbance measurement, 460 nm) in the presence of organic solvent	Acetone	60%	100	0	%	glycine 100 mM	8.6	37	? mM 1-Iodohexane	1-Hexanol, Iodide ion	—	—	Classical aqueous control (in %)
—	Activity - Classical	Activity measured by absorbance spectrophotometry (colorimetric assay, iodine, mercuric thiocyanate and ferric ammonium sulfate reaction product Fe–SCN absorbance measurement, 460 nm) in the presence of organic solvent	Isopropanol	60%	100	0	%	glycine 100 mM	8.6	37	? mM 1-Iodohexane	1-Hexanol, Iodide ion	—	—	Classical aqueous control (in %)
—	Activity - Classical	Activity measured by absorbance spectrophotometry (colorimetric assay, iodine, mercuric thiocyanate and ferric ammonium sulfate reaction product Fe–SCN absorbance measurement, 460 nm) in the presence of organic solvent	Tetrahydrofuran (THF)	60%	100	0	%	glycine 100 mM	8.6	37	? mM 1-Iodohexane	1-Hexanol, Iodide ion	—	—	Classical aqueous control (in %)
—	Activity - Classical	Activity measured by absorbance spectrophotometry (colorimetric assay, iodine, mercuric thiocyanate and ferric ammonium sulfate reaction product Fe–SCN absorbance measurement, 460 nm) in the presence of organic solvent	Glycerol	75%	100	65	%	glycine 100 mM	8.6	37	? mM 1-Iodohexane	1-Hexanol, Iodide ion	—	—	Classical aqueous control (in %)
—	Activity - Classical	Activity measured by absorbance spectrophotometry (colorimetric assay, iodine, mercuric thiocyanate and ferric ammonium sulfate reaction product Fe–SCN absorbance measurement, 460 nm) in the presence of organic solvent	Formamide	75%	100	0	%	glycine 100 mM	8.6	37	? mM 1-Iodohexane	1-Hexanol, Iodide ion	—	—	Classical aqueous control (in %)
—	Activity - Classical	Activity measured by absorbance spectrophotometry (colorimetric assay, iodine, mercuric thiocyanate and ferric ammonium sulfate reaction product Fe–SCN absorbance measurement, 460 nm) in the presence of organic solvent	Ethylene Glycol	75%	100	4	%	glycine 100 mM	8.6	37	? mM 1-Iodohexane	1-Hexanol, Iodide ion	—	—	Classical aqueous control (in %)
—	Activity - Classical	Activity measured by absorbance spectrophotometry (colorimetric assay, iodine, mercuric thiocyanate and ferric ammonium sulfate reaction product Fe–SCN absorbance measurement, 460 nm) in the presence of organic solvent	PEG-1000	75%	100	125	%	glycine 100 mM	8.6	37	? mM 1-Iodohexane	1-Hexanol, Iodide ion	—	—	Classical aqueous control (in %)
—	Activity - Classical	Activity measured by absorbance spectrophotometry (colorimetric assay, iodine, mercuric thiocyanate and ferric ammonium sulfate reaction product Fe–SCN absorbance measurement, 460 nm) in the presence of organic solvent	PEG-10000	75%	100	0	%	glycine 100 mM	8.6	37	? mM 1-Iodohexane	1-Hexanol, Iodide ion	—	—	Classical aqueous control (in %)
—	Activity - Classical	Activity measured by absorbance spectrophotometry (colorimetric assay, iodine, mercuric thiocyanate and ferric ammonium sulfate reaction product Fe–SCN absorbance measurement, 460 nm) in the presence of organic solvent	Dimethylformamide (DMF)	75%	100	0	%	glycine 100 mM	8.6	37	? mM 1-Iodohexane	1-Hexanol, Iodide ion	—	—	Classical aqueous control (in %)
—	Activity - Classical	Activity measured by absorbance spectrophotometry (colorimetric assay, iodine, mercuric thiocyanate and ferric ammonium sulfate reaction product Fe–SCN absorbance measurement, 460 nm) in the presence of organic solvent	Dimethyl Sulfoxide (DMSO)	75%	100	0	%	glycine 100 mM	8.6	37	? mM 1-Iodohexane	1-Hexanol, Iodide ion	—	—	Classical aqueous control (in %)
—	Activity - Classical	Activity measured by absorbance spectrophotometry (colorimetric assay, iodine, mercuric thiocyanate and ferric ammonium sulfate reaction product Fe–SCN absorbance measurement, 460 nm) in the presence of organic solvent	Methanol	75%	100	0	%	glycine 100 mM	8.6	37	? mM 1-Iodohexane	1-Hexanol, Iodide ion	—	—	Classical aqueous control (in %)
—	Activity - Classical	Activity measured by absorbance spectrophotometry (colorimetric assay, iodine, mercuric thiocyanate and ferric ammonium sulfate reaction product Fe–SCN absorbance measurement, 460 nm) in the presence of organic solvent	1,4-Dioxane	75%	100	0	%	glycine 100 mM	8.6	37	? mM 1-Iodohexane	1-Hexanol, Iodide ion	—	—	Classical aqueous control (in %)
—	Activity - Classical	Activity measured by absorbance spectrophotometry (colorimetric assay, iodine, mercuric thiocyanate and ferric ammonium sulfate reaction product Fe–SCN absorbance measurement, 460 nm) in the presence of organic solvent	Acetonitrile	75%	100	0	%	glycine 100 mM	8.6	37	? mM 1-Iodohexane	1-Hexanol, Iodide ion	—	—	Classical aqueous control (in %)
—	Activity - Classical	Activity measured by absorbance spectrophotometry (colorimetric assay, iodine, mercuric thiocyanate and ferric ammonium sulfate reaction product Fe–SCN absorbance measurement, 460 nm) in the presence of organic solvent	Ethanol	75%	100	0	%	glycine 100 mM	8.6	37	? mM 1-Iodohexane	1-Hexanol, Iodide ion	—	—	Classical aqueous control (in %)
—	Activity - Classical	Activity measured by absorbance spectrophotometry (colorimetric assay, iodine, mercuric thiocyanate and ferric ammonium sulfate reaction product Fe–SCN absorbance measurement, 460 nm) in the presence of organic solvent	Acetone	75%	100	0	%	glycine 100 mM	8.6	37	? mM 1-Iodohexane	1-Hexanol, Iodide ion	—	—	Classical aqueous control (in %)
—	Activity - Classical	Activity measured by absorbance spectrophotometry (colorimetric assay, iodine, mercuric thiocyanate and ferric ammonium sulfate reaction product Fe–SCN absorbance measurement, 460 nm) in the presence of organic solvent	Isopropanol	75%	100	0	%	glycine 100 mM	8.6	37	? mM 1-Iodohexane	1-Hexanol, Iodide ion	—	—	Classical aqueous control (in %)
—	Activity - Classical	Activity measured by absorbance spectrophotometry (colorimetric assay, iodine, mercuric thiocyanate and ferric ammonium sulfate reaction product Fe–SCN absorbance measurement, 460 nm) in the presence of organic solvent	Tetrahydrofuran (THF)	75%	100	0	%	glycine 100 mM	8.6	37	? mM 1-Iodohexane	1-Hexanol, Iodide ion	—	—	Classical aqueous control (in %)
—	Stability - C50	Volume fraction of organic solvent at which 50% of enzyme activity is lost, measured by absorbance spectrophotometry (colorimetric assay, iodine, mercuric thiocyanate and ferric ammonium sulfate reaction product Fe–SCN absorbance measurement, 460 nm) in the presence of organic solvent	Glycerol	—	—	75	% (v/v)	glycine 100 mM	8.6	37	1-Iodohexane	1-Hexanol, Iodide ion	—	—	Not applicable control (in % (v/v))
—	Stability - C50	Volume fraction of organic solvent at which 50% of enzyme activity is lost, measured by absorbance spectrophotometry (colorimetric assay, iodine, mercuric thiocyanate and ferric ammonium sulfate reaction product Fe–SCN absorbance measurement, 460 nm) in the presence of organic solvent	Formamide	—	—	30	% (v/v)	glycine 100 mM	8.6	37	1-Iodohexane	1-Hexanol, Iodide ion	—	—	Not applicable control (in % (v/v))
—	Stability - C50	Volume fraction of organic solvent at which 50% of enzyme activity is lost, measured by absorbance spectrophotometry (colorimetric assay, iodine, mercuric thiocyanate and ferric ammonium sulfate reaction product Fe–SCN absorbance measurement, 460 nm) in the presence of organic solvent	Ethylene Glycol	—	—	62	% (v/v)	glycine 100 mM	8.6	37	1-Iodohexane	1-Hexanol, Iodide ion	—	—	Not applicable control (in % (v/v))
—	Stability - C50	Volume fraction of organic solvent at which 50% of enzyme activity is lost, measured by absorbance spectrophotometry (colorimetric assay, iodine, mercuric thiocyanate and ferric ammonium sulfate reaction product Fe–SCN absorbance measurement, 460 nm) in the presence of organic solvent	Dimethyl Sulfoxide (DMSO)	—	—	48	% (v/v)	glycine 100 mM	8.6	37	1-Iodohexane	1-Hexanol, Iodide ion	—	—	Not applicable control (in % (v/v))
—	Stability - C50	Volume fraction of organic solvent at which 50% of enzyme activity is lost, measured by absorbance spectrophotometry (colorimetric assay, iodine, mercuric thiocyanate and ferric ammonium sulfate reaction product Fe–SCN absorbance measurement, 460 nm) in the presence of organic solvent	Dimethylformamide (DMF)	—	—	23	% (v/v)	glycine 100 mM	8.6	37	1-Iodohexane	1-Hexanol, Iodide ion	—	—	Not applicable control (in % (v/v))
—	Stability - C50	Volume fraction of organic solvent at which 50% of enzyme activity is lost, measured by absorbance spectrophotometry (colorimetric assay, iodine, mercuric thiocyanate and ferric ammonium sulfate reaction product Fe–SCN absorbance measurement, 460 nm) in the presence of organic solvent	Methanol	—	—	25	% (v/v)	glycine 100 mM	8.6	37	1-Iodohexane	1-Hexanol, Iodide ion	—	—	Not applicable control (in % (v/v))
—	Stability - C50	Volume fraction of organic solvent at which 50% of enzyme activity is lost, measured by absorbance spectrophotometry (colorimetric assay, iodine, mercuric thiocyanate and ferric ammonium sulfate reaction product Fe–SCN absorbance measurement, 460 nm) in the presence of organic solvent	Acetonitrile	—	—	9	% (v/v)	glycine 100 mM	8.6	37	1-Iodohexane	1-Hexanol, Iodide ion	—	—	Not applicable control (in % (v/v))
—	Stability - C50	Volume fraction of organic solvent at which 50% of enzyme activity is lost, measured by absorbance spectrophotometry (colorimetric assay, iodine, mercuric thiocyanate and ferric ammonium sulfate reaction product Fe–SCN absorbance measurement, 460 nm) in the presence of organic solvent	Ethanol	—	—	22	% (v/v)	glycine 100 mM	8.6	37	1-Iodohexane	1-Hexanol, Iodide ion	—	—	Not applicable control (in % (v/v))
—	Stability - C50	Volume fraction of organic solvent at which 50% of enzyme activity is lost, measured by absorbance spectrophotometry (colorimetric assay, iodine, mercuric thiocyanate and ferric ammonium sulfate reaction product Fe–SCN absorbance measurement, 460 nm) in the presence of organic solvent	1,4-Dioxane	—	—	4	% (v/v)	glycine 100 mM	8.6	37	1-Iodohexane	1-Hexanol, Iodide ion	—	—	Not applicable control (in % (v/v))
—	Stability - C50	Volume fraction of organic solvent at which 50% of enzyme activity is lost, measured by absorbance spectrophotometry (colorimetric assay, iodine, mercuric thiocyanate and ferric ammonium sulfate reaction product Fe–SCN absorbance measurement, 460 nm) in the presence of organic solvent	Acetone	—	—	16	% (v/v)	glycine 100 mM	8.6	37	1-Iodohexane	1-Hexanol, Iodide ion	—	—	Not applicable control (in % (v/v))
—	Stability - C50	Volume fraction of organic solvent at which 50% of enzyme activity is lost, measured by absorbance spectrophotometry (colorimetric assay, iodine, mercuric thiocyanate and ferric ammonium sulfate reaction product Fe–SCN absorbance measurement, 460 nm) in the presence of organic solvent	Isopropanol	—	—	9	% (v/v)	glycine 100 mM	8.6	37	1-Iodohexane	1-Hexanol, Iodide ion	—	—	Not applicable control (in % (v/v))
—	Stability - C50	Volume fraction of organic solvent at which 50% of enzyme activity is lost, measured by absorbance spectrophotometry (colorimetric assay, iodine, mercuric thiocyanate and ferric ammonium sulfate reaction product Fe–SCN absorbance measurement, 460 nm) in the presence of organic solvent	Tetrahydrofuran (THF)	—	—	7	% (v/v)	glycine 100 mM	8.6	37	1-Iodohexane	1-Hexanol, Iodide ion	—	—	Not applicable control (in % (v/v))
—	Stability - Tm	Tm measured by circular dichroism in the presence of organic solvent	Ethylene Glycol	20%	51	50	°C	50 mM phosphate buffer	7.5	—	—	—	—	—	Classical aqueous control (in °C), averaged between circular dichroism and DSC measurements
—	Stability - Tm	Tm measured by DSC in the presence of organic solvent	Dimethylformamide (DMF)	20%	51	34	°C	50 mM phosphate buffer	7.5	—	—	—	—	—	Classical aqueous control (in °C), averaged between circular dichroism and DSC measurements
—	Stability - Tm	Tm measured by DSC in the presence of organic solvent	Dimethyl Sulfoxide (DMSO)	20%	51	47	°C	50 mM phosphate buffer	7.5	—	—	—	—	—	Classical aqueous control (in °C), averaged between circular dichroism and DSC measurements
—	Stability - Tm	Tm measured by circular dichroism in the presence of organic solvent	Methanol	20%	51	40	°C	50 mM phosphate buffer	7.5	—	—	—	—	—	Classical aqueous control (in °C), averaged between circular dichroism and DSC measurements
—	Stability - Tm	Tm measured by circular dichroism in the presence of organic solvent	1,4-Dioxane	20%	51	30	°C	50 mM phosphate buffer	7.5	—	—	—	—	—	Classical aqueous control (in °C), averaged between circular dichroism and DSC measurements
—	Stability - Tm	Tm measured by circular dichroism in the presence of organic solvent	Acetonitrile	10%	51	37	°C	50 mM phosphate buffer	7.5	—	—	—	—	—	Classical aqueous control (in °C), averaged between circular dichroism and DSC measurements
—	Stability - Tm	Tm measured by circular dichroism in the presence of organic solvent	Acetonitrile	20%	51	<10	°C	50 mM phosphate buffer	7.5	—	—	—	—	—	Classical aqueous control (in °C), averaged between circular dichroism and DSC measurements
—	Stability - Tm	Tm measured by circular dichroism in the presence of organic solvent	Ethanol	20%	51	35	°C	50 mM phosphate buffer	7.5	—	—	—	—	—	Classical aqueous control (in °C), averaged between circular dichroism and DSC measurements
—	Stability - Tm	Tm measured by circular dichroism in the presence of organic solvent	Acetone	20%	51	31	°C	50 mM phosphate buffer	7.5	—	—	—	—	—	Classical aqueous control (in °C), averaged between circular dichroism and DSC measurements
—	Stability - Tm	Tm measured by circular dichroism in the presence of organic solvent	Isopropanol	20%	51	33	°C	50 mM phosphate buffer	7.5	—	—	—	—	—	Classical aqueous control (in °C), averaged between circular dichroism and DSC measurements
—	Stability - Tm	Tm measured by circular dichroism in the presence of organic solvent	Tetrahydrofuran (THF)	10%	51	32	°C	50 mM phosphate buffer	7.5	—	—	—	—	—	Classical aqueous control (in °C), averaged between circular dichroism and DSC measurements
—	Stability - Tm	Tm measured by circular dichroism in the presence of organic solvent	Tetrahydrofuran (THF)	20%	51	0	°C	50 mM phosphate buffer	7.5	—	—	—	—	—	Classical aqueous control (in °C), averaged between circular dichroism and DSC measurements

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Visualization : Activity — Classical

Veronika Stepankova et al. (2013)

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

Visualization : Stability — T_m

Veronika Stepankova et al. (2013)

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

Visualization : Stability — C₅₀

Veronika Stepankova et al. (2013)

One bar per measurement. Colour = solvent, shade = temperature. Hover for details.

Tana Koudelakova et al. (2013) — Engineering Enzyme Stability and Resistance to an Organic Cosolvent by Modification of Residues in the Access Tunnel

Angewandte Chemie International Edition · doi:10.1002/anie.201206708 ↗ · Activity - Classical Activity - Michaelis-Menten Stability - C50 Stability - Inactivation Rate Constant Stability - Tm

66 measurements ▼

Database ID

UniProt: P0A3G2 ↗

Sequence Annotation

Explicit - Provided PDB Accession Number

Protein Source

Recombinant, host bacterium Escherichia coli BL21

Experimental Data (66 measurements)

66 measurements — page 1 of 4

Mutation	Property	Assay	Solvent	Solvent Volume	Aqueous Reference	WT Reference	Measured Value	Units	Solution	pH	Temperature	Substrate(s)	Product(s)	Cofactor(s)	Shaking	Comments
WT	Activity - Classical	Activity (in nmol/(s*mg)), measured by absorbance spectrophotometry (colorimetric assay, Br ion, mercuric thiocyanate and ferric ammonium sulfate reaction product absorbance measurement, 460 nm) in the presence of organic solvent	Dimethyl Sulfoxide (DMSO)	40%	100.0	—	2.19	%	100 mM glycine buffer	8.6	—	1,2-Dibromoethane	2-Bromoethanol , Bromide ion	—	—	Classical aqueous control (in %)
WT	Activity - Michaelis-Menten	kcat (1/s) measured by absorbance spectrophotometry (colorimetric assay, Br ion, mercuric thiocyanate and ferric ammonium sulfate reaction product absorbance measurement, 460 nm) in the presence of organic solvent	Dimethyl Sulfoxide (DMSO)	40%	28.67	—	2.25	1/s	100 mM glycine buffer	8.6	37°C	1,2-Dibromoethane	2-Bromoethanol , Bromide ion	—	—	Classical aqueous control (in 1/s)
WT	Activity - Michaelis-Menten	kcat/Km (1/(s*mM)) measured by absorbance spectrophotometry (colorimetric assay, Br ion, mercuric thiocyanate and ferric ammonium sulfate reaction product absorbance measurement, 460 nm) in the presence of organic solvent	Dimethyl Sulfoxide (DMSO)	40%	8.06	—	0.16	1/(s * mM)	100 mM glycine buffer	8.6	37°C	1,2-Dibromoethane	2-Bromoethanol , Bromide ion	—	—	Classical aqueous control (in 1/(s * mM))
WT	Activity - Michaelis-Menten	Km (mM) measured by absorbance spectrophotometry (colorimetric assay, Br ion, mercuric thiocyanate and ferric ammonium sulfate reaction product absorbance measurement, 460 nm) in the presence of organic solvent	Dimethyl Sulfoxide (DMSO)	40%	3.56	—	14.17	mM	100 mM glycine buffer	8.6	37°C	1,2-Dibromoethane	2-Bromoethanol , Bromide ion	—	—	Classical aqueous control (in mM)
WT	Stability - C50	Volume percentage of DMSO (Dimethyl Sulfoxide) for which half enzyme is unfolded after 30 minutes incubation at 37°C in the presence of organic solvent, measured by fluorescence spectrophotometry	Dimethyl Sulfoxide (DMSO)	—	—	—	29.1	%	50 mM phosphate buffer	7.5	37°C	—	—	—	—	Classical aqueous control (in %)
WT	Stability - C50	Volume percentage of DMSO (Dimethyl Sulfoxide) for which half enzyme is unfolded at 23°C, measured by fluorescence spectrophotometry	Dimethyl Sulfoxide (DMSO)	—	—	—	49.0	%	50 mM phosphate buffer	7.5	23°C	—	—	—	—	Classical aqueous control (in %)
WT	Stability - Inactivation Rate Constant	Ksi (mM) measured by absorbance spectrophotometry (colorimetric assay, Br ion, mercuric thiocyanate and ferric ammonium sulfate reaction product absorbance measurement, 460 nm) in the presence of organic solvent	Dimethyl Sulfoxide (DMSO)	40%	3.56	—	0.0	mM	100 mM glycine buffer	8.6	37°C	1,2-Dibromoethane	2-Bromoethanol , Bromide ion	—	—	Classical aqueous control (in mM)
WT	Stability - Tm	Thermal stability measured by DSC in the presence of organic solvent	Dimethyl Sulfoxide (DMSO)	40%	51.9	—	39.0	°C	50 mM phosphate buffer	7.5	—	—	—	—	—	Classical aqueous control (in °C)
WT	Stability - Tm	Thermal stability measured by DSC in the presence of organic solvent	Acetonitrile	10%	51.9	—	36.8	°C	50 mM phosphate buffer	7.5	—	—	—	—	—	Classical aqueous control (in °C)
WT	Stability - Tm	Thermal stability measured by DSC in the presence of organic solvent	tert-Butanol	20%	51.9	—	30.6	°C	50 mM phosphate buffer	7.5	—	—	—	—	—	Classical aqueous control (in °C)
F144I/C176G	Activity - Classical	Activity (in nmol/(s*mg)), measured by absorbance spectrophotometry (colorimetric assay, Br ion, mercuric thiocyanate and ferric ammonium sulfate reaction product absorbance measurement, 460 nm) in the presence of organic solvent	Dimethyl Sulfoxide (DMSO)	40%	100.0	2.19	0.44	%	100 mM glycine buffer	8.6	—	1,2-Dibromoethane	2-Bromoethanol , Bromide ion	—	—	Classical aqueous control (in %)
D73G/E98V	Activity - Classical	Activity (in nmol/(s*mg)), measured by absorbance spectrophotometry (colorimetric assay, Br ion, mercuric thiocyanate and ferric ammonium sulfate reaction product absorbance measurement, 460 nm) in the presence of organic solvent	Dimethyl Sulfoxide (DMSO)	40%	100.0	2.19	1.46	%	100 mM glycine buffer	8.6	—	1,2-Dibromoethane	2-Bromoethanol , Bromide ion	—	—	Classical aqueous control (in %)
V245L	Activity - Classical	Activity (in nmol/(s*mg)), measured by absorbance spectrophotometry (colorimetric assay, Br ion, mercuric thiocyanate and ferric ammonium sulfate reaction product absorbance measurement, 460 nm) in the presence of organic solvent	Dimethyl Sulfoxide (DMSO)	40%	100.0	2.19	0.66	%	100 mM glycine buffer	8.6	—	1,2-Dibromoethane	2-Bromoethanol , Bromide ion	—	—	Classical aqueous control (in %)
L95V/A172V	Activity - Classical	Activity (in nmol/(s*mg)), measured by absorbance spectrophotometry (colorimetric assay, Br ion, mercuric thiocyanate and ferric ammonium sulfate reaction product absorbance measurement, 460 nm) in the presence of organic solvent	Dimethyl Sulfoxide (DMSO)	40%	100.0	2.19	10.4	%	100 mM glycine buffer	8.6	—	1,2-Dibromoethane	2-Bromoethanol , Bromide ion	—	—	Classical aqueous control (in %)
T148L/G171Q/A172V/C176F/	Activity - Michaelis-Menten	kcat (1/s) measured by absorbance spectrophotometry (colorimetric assay, Br ion, mercuric thiocyanate and ferric ammonium sulfate reaction product absorbance measurement, 460 nm) in the presence of organic solvent	Dimethyl Sulfoxide (DMSO)	40%	0.34	2.25	0.25	1/s	100 mM glycine buffer	8.6	37°C	1,2-Dibromoethane	2-Bromoethanol , Bromide ion	—	—	Classical aqueous control (in 1/s)
L95V/A172V	Activity - Michaelis-Menten	kcat (1/s) measured by absorbance spectrophotometry (colorimetric assay, Br ion, mercuric thiocyanate and ferric ammonium sulfate reaction product absorbance measurement, 460 nm) in the presence of organic solvent	Dimethyl Sulfoxide (DMSO)	40%	9.88	2.25	0.5	1/s	100 mM glycine buffer	8.6	37°C	1,2-Dibromoethane	2-Bromoethanol , Bromide ion	—	—	Classical aqueous control (in 1/s)
A172V	Activity - Michaelis-Menten	kcat (1/s) measured by absorbance spectrophotometry (colorimetric assay, Br ion, mercuric thiocyanate and ferric ammonium sulfate reaction product absorbance measurement, 460 nm) in the presence of organic solvent	Dimethyl Sulfoxide (DMSO)	40%	13.83	2.25	1.01	1/s	100 mM glycine buffer	8.6	37°C	1,2-Dibromoethane	2-Bromoethanol , Bromide ion	—	—	Classical aqueous control (in 1/s)
D78G/F80S/T148L/G171Q/A172V/C176F/N227T/W240Y/P291A/A292G	Activity - Michaelis-Menten	kcat (1/s) measured by absorbance spectrophotometry (colorimetric assay, Br ion, mercuric thiocyanate and ferric ammonium sulfate reaction product absorbance measurement, 460 nm) in the presence of organic solvent	Dimethyl Sulfoxide (DMSO)	40%	2.23	2.25	0.72	1/s	100 mM glycine buffer	8.6	37°C	1,2-Dibromoethane	2-Bromoethanol , Bromide ion	—	—	Classical aqueous control (in 1/s)
T148L/G171V/A172VC176F	Activity - Michaelis-Menten	kcat (1/s) measured by absorbance spectrophotometry (colorimetric assay, Br ion, mercuric thiocyanate and ferric ammonium sulfate reaction product absorbance measurement, 460 nm) in the presence of organic solvent	Dimethyl Sulfoxide (DMSO)	40%	0.26	2.25	0.36	1/s	100 mM glycine buffer	8.6	37°C	1,2-Dibromoethane	2-Bromoethanol , Bromide ion	—	—	Classical aqueous control (in 1/s)
T148L/G171L/A172VC176F	Activity - Michaelis-Menten	kcat (1/s) measured by absorbance spectrophotometry (colorimetric assay, Br ion, mercuric thiocyanate and ferric ammonium sulfate reaction product absorbance measurement, 460 nm) in the presence of organic solvent	Dimethyl Sulfoxide (DMSO)	40%	0.28	2.25	0.86	1/s	100 mM glycine buffer	8.6	37°C	1,2-Dibromoethane	2-Bromoethanol , Bromide ion	—	—	Classical aqueous control (in 1/s)

WT	Activity - Classical	Activity (in nmol/(s*mg)), measured by absorbance spectrophotometry (colorimetric assay, Br ion, mercuric thiocyanate and ferric ammonium sulfate reaction product absorbance measurement, 460 nm) in the presence of organic solvent	Dimethyl Sulfoxide (DMSO)	40%	100.0	—	2.19	%	100 mM glycine buffer	8.6	—	1,2-Dibromoethane	2-Bromoethanol, Bromide ion	—	—	Classical aqueous control (in %)
WT	Activity - Michaelis-Menten	kcat (1/s) measured by absorbance spectrophotometry (colorimetric assay, Br ion, mercuric thiocyanate and ferric ammonium sulfate reaction product absorbance measurement, 460 nm) in the presence of organic solvent	Dimethyl Sulfoxide (DMSO)	40%	28.67	—	2.25	1/s	100 mM glycine buffer	8.6	37°C	1,2-Dibromoethane	2-Bromoethanol, Bromide ion	—	—	Classical aqueous control (in 1/s)
WT	Activity - Michaelis-Menten	kcat/Km (1/(s*mM)) measured by absorbance spectrophotometry (colorimetric assay, Br ion, mercuric thiocyanate and ferric ammonium sulfate reaction product absorbance measurement, 460 nm) in the presence of organic solvent	Dimethyl Sulfoxide (DMSO)	40%	8.06	—	0.16	1/(s * mM)	100 mM glycine buffer	8.6	37°C	1,2-Dibromoethane	2-Bromoethanol, Bromide ion	—	—	Classical aqueous control (in 1/(s * mM))
WT	Activity - Michaelis-Menten	Km (mM) measured by absorbance spectrophotometry (colorimetric assay, Br ion, mercuric thiocyanate and ferric ammonium sulfate reaction product absorbance measurement, 460 nm) in the presence of organic solvent	Dimethyl Sulfoxide (DMSO)	40%	3.56	—	14.17	mM	100 mM glycine buffer	8.6	37°C	1,2-Dibromoethane	2-Bromoethanol, Bromide ion	—	—	Classical aqueous control (in mM)
WT	Stability - C50	Volume percentage of DMSO (Dimethyl Sulfoxide) for which half enzyme is unfolded after 30 minutes incubation at 37°C in the presence of organic solvent, measured by fluorescence spectrophotometry	Dimethyl Sulfoxide (DMSO)	—	—	—	29.1	%	50 mM phosphate buffer	7.5	37°C	—	—	—	—	Classical aqueous control (in %)
WT	Stability - C50	Volume percentage of DMSO (Dimethyl Sulfoxide) for which half enzyme is unfolded at 23°C, measured by fluorescence spectrophotometry	Dimethyl Sulfoxide (DMSO)	—	—	—	49.0	%	50 mM phosphate buffer	7.5	23°C	—	—	—	—	Classical aqueous control (in %)
WT	Stability - Inactivation Rate Constant	Ksi (mM) measured by absorbance spectrophotometry (colorimetric assay, Br ion, mercuric thiocyanate and ferric ammonium sulfate reaction product absorbance measurement, 460 nm) in the presence of organic solvent	Dimethyl Sulfoxide (DMSO)	40%	3.56	—	0.0	mM	100 mM glycine buffer	8.6	37°C	1,2-Dibromoethane	2-Bromoethanol, Bromide ion	—	—	Classical aqueous control (in mM)
WT	Stability - Tm	Thermal stability measured by DSC in the presence of organic solvent	Dimethyl Sulfoxide (DMSO)	40%	51.9	—	39.0	°C	50 mM phosphate buffer	7.5	—	—	—	—	—	Classical aqueous control (in °C)
WT	Stability - Tm	Thermal stability measured by DSC in the presence of organic solvent	Acetonitrile	10%	51.9	—	36.8	°C	50 mM phosphate buffer	7.5	—	—	—	—	—	Classical aqueous control (in °C)
WT	Stability - Tm	Thermal stability measured by DSC in the presence of organic solvent	tert-Butanol	20%	51.9	—	30.6	°C	50 mM phosphate buffer	7.5	—	—	—	—	—	Classical aqueous control (in °C)
F144I/C176G	Activity - Classical	Activity (in nmol/(s*mg)), measured by absorbance spectrophotometry (colorimetric assay, Br ion, mercuric thiocyanate and ferric ammonium sulfate reaction product absorbance measurement, 460 nm) in the presence of organic solvent	Dimethyl Sulfoxide (DMSO)	40%	100.0	2.19	0.44	%	100 mM glycine buffer	8.6	—	1,2-Dibromoethane	2-Bromoethanol, Bromide ion	—	—	Classical aqueous control (in %)
D73G/E98V	Activity - Classical	Activity (in nmol/(s*mg)), measured by absorbance spectrophotometry (colorimetric assay, Br ion, mercuric thiocyanate and ferric ammonium sulfate reaction product absorbance measurement, 460 nm) in the presence of organic solvent	Dimethyl Sulfoxide (DMSO)	40%	100.0	2.19	1.46	%	100 mM glycine buffer	8.6	—	1,2-Dibromoethane	2-Bromoethanol, Bromide ion	—	—	Classical aqueous control (in %)
V245L	Activity - Classical	Activity (in nmol/(s*mg)), measured by absorbance spectrophotometry (colorimetric assay, Br ion, mercuric thiocyanate and ferric ammonium sulfate reaction product absorbance measurement, 460 nm) in the presence of organic solvent	Dimethyl Sulfoxide (DMSO)	40%	100.0	2.19	0.66	%	100 mM glycine buffer	8.6	—	1,2-Dibromoethane	2-Bromoethanol, Bromide ion	—	—	Classical aqueous control (in %)
L95V/A172V	Activity - Classical	Activity (in nmol/(s*mg)), measured by absorbance spectrophotometry (colorimetric assay, Br ion, mercuric thiocyanate and ferric ammonium sulfate reaction product absorbance measurement, 460 nm) in the presence of organic solvent	Dimethyl Sulfoxide (DMSO)	40%	100.0	2.19	10.4	%	100 mM glycine buffer	8.6	—	1,2-Dibromoethane	2-Bromoethanol, Bromide ion	—	—	Classical aqueous control (in %)
T148L/G171Q/A172V/C176F/	Activity - Michaelis-Menten	kcat (1/s) measured by absorbance spectrophotometry (colorimetric assay, Br ion, mercuric thiocyanate and ferric ammonium sulfate reaction product absorbance measurement, 460 nm) in the presence of organic solvent	Dimethyl Sulfoxide (DMSO)	40%	0.34	2.25	0.25	1/s	100 mM glycine buffer	8.6	37°C	1,2-Dibromoethane	2-Bromoethanol, Bromide ion	—	—	Classical aqueous control (in 1/s)
L95V/A172V	Activity - Michaelis-Menten	kcat (1/s) measured by absorbance spectrophotometry (colorimetric assay, Br ion, mercuric thiocyanate and ferric ammonium sulfate reaction product absorbance measurement, 460 nm) in the presence of organic solvent	Dimethyl Sulfoxide (DMSO)	40%	9.88	2.25	0.5	1/s	100 mM glycine buffer	8.6	37°C	1,2-Dibromoethane	2-Bromoethanol, Bromide ion	—	—	Classical aqueous control (in 1/s)
A172V	Activity - Michaelis-Menten	kcat (1/s) measured by absorbance spectrophotometry (colorimetric assay, Br ion, mercuric thiocyanate and ferric ammonium sulfate reaction product absorbance measurement, 460 nm) in the presence of organic solvent	Dimethyl Sulfoxide (DMSO)	40%	13.83	2.25	1.01	1/s	100 mM glycine buffer	8.6	37°C	1,2-Dibromoethane	2-Bromoethanol, Bromide ion	—	—	Classical aqueous control (in 1/s)
D78G/F80S/T148L/G171Q/A172V/C176F/N227T/W240Y/P291A/A292G	Activity - Michaelis-Menten	kcat (1/s) measured by absorbance spectrophotometry (colorimetric assay, Br ion, mercuric thiocyanate and ferric ammonium sulfate reaction product absorbance measurement, 460 nm) in the presence of organic solvent	Dimethyl Sulfoxide (DMSO)	40%	2.23	2.25	0.72	1/s	100 mM glycine buffer	8.6	37°C	1,2-Dibromoethane	2-Bromoethanol, Bromide ion	—	—	Classical aqueous control (in 1/s)
T148L/G171V/A172VC176F	Activity - Michaelis-Menten	kcat (1/s) measured by absorbance spectrophotometry (colorimetric assay, Br ion, mercuric thiocyanate and ferric ammonium sulfate reaction product absorbance measurement, 460 nm) in the presence of organic solvent	Dimethyl Sulfoxide (DMSO)	40%	0.26	2.25	0.36	1/s	100 mM glycine buffer	8.6	37°C	1,2-Dibromoethane	2-Bromoethanol, Bromide ion	—	—	Classical aqueous control (in 1/s)
T148L/G171L/A172VC176F	Activity - Michaelis-Menten	kcat (1/s) measured by absorbance spectrophotometry (colorimetric assay, Br ion, mercuric thiocyanate and ferric ammonium sulfate reaction product absorbance measurement, 460 nm) in the presence of organic solvent	Dimethyl Sulfoxide (DMSO)	40%	0.28	2.25	0.86	1/s	100 mM glycine buffer	8.6	37°C	1,2-Dibromoethane	2-Bromoethanol, Bromide ion	—	—	Classical aqueous control (in 1/s)
L95V/A172V	Activity - Michaelis-Menten	kcat/Km (1/(s*mM)) measured by absorbance spectrophotometry (colorimetric assay, Br ion, mercuric thiocyanate and ferric ammonium sulfate reaction product absorbance measurement, 460 nm) in the presence of organic solvent	Dimethyl Sulfoxide (DMSO)	40%	11.66	0.16	0.12	1/(s * mM)	100 mM glycine buffer	8.6	37°C	1,2-Dibromoethane	2-Bromoethanol, Bromide ion	—	—	Classical aqueous control (in 1/(s * mM))
A172V	Activity - Michaelis-Menten	kcat/Km (1/(s*mM)) measured by absorbance spectrophotometry (colorimetric assay, Br ion, mercuric thiocyanate and ferric ammonium sulfate reaction product absorbance measurement, 460 nm) in the presence of organic solvent	Dimethyl Sulfoxide (DMSO)	40%	11.79	0.16	0.18	1/(s * mM)	100 mM glycine buffer	8.6	37°C	1,2-Dibromoethane	2-Bromoethanol, Bromide ion	—	—	Classical aqueous control (in 1/(s * mM))
D78G/F80S/T148L/G171Q/A172V/C176F/N227T/W240Y/P291A/A292G	Activity - Michaelis-Menten	kcat/Km (1/(s*mM)) measured by absorbance spectrophotometry (colorimetric assay, Br ion, mercuric thiocyanate and ferric ammonium sulfate reaction product absorbance measurement, 460 nm) in the presence of organic solvent	Dimethyl Sulfoxide (DMSO)	40%	1.31	0.16	0.67	1/(s * mM)	100 mM glycine buffer	8.6	37°C	1,2-Dibromoethane	2-Bromoethanol, Bromide ion	—	—	Classical aqueous control (in 1/(s * mM))
T148L/G171Q/A172V/C176F/	Activity - Michaelis-Menten	kcat/Km (1/(s*mM)) measured by absorbance spectrophotometry (colorimetric assay, Br ion, mercuric thiocyanate and ferric ammonium sulfate reaction product absorbance measurement, 460 nm) in the presence of organic solvent	Dimethyl Sulfoxide (DMSO)	40%	2.49	0.16	0.29	1/(s * mM)	100 mM glycine buffer	8.6	37°C	1,2-Dibromoethane	2-Bromoethanol, Bromide ion	—	—	Classical aqueous control (in 1/(s * mM))
T148L/G171V/A172VC176F	Activity - Michaelis-Menten	kcat/Km (1/(s*mM)) measured by absorbance spectrophotometry (colorimetric assay, Br ion, mercuric thiocyanate and ferric ammonium sulfate reaction product absorbance measurement, 460 nm) in the presence of organic solvent	Dimethyl Sulfoxide (DMSO)	40%	2.46	0.16	0.26	1/(s * mM)	100 mM glycine buffer	8.6	37°C	1,2-Dibromoethane	2-Bromoethanol, Bromide ion	—	—	Classical aqueous control (in 1/(s * mM))
T148L/G171L/A172VC176F	Activity - Michaelis-Menten	kcat/Km (1/(s*mM)) measured by absorbance spectrophotometry (colorimetric assay, Br ion, mercuric thiocyanate and ferric ammonium sulfate reaction product absorbance measurement, 460 nm) in the presence of organic solvent	Dimethyl Sulfoxide (DMSO)	40%	2.45	0.16	0.17	1/(s * mM)	100 mM glycine buffer	8.6	37°C	1,2-Dibromoethane	2-Bromoethanol, Bromide ion	—	—	Classical aqueous control (in 1/(s * mM))
T148L/G171L/A172VC176F	Activity - Michaelis-Menten	Km (mM) measured by absorbance spectrophotometry (colorimetric assay, Br ion, mercuric thiocyanate and ferric ammonium sulfate reaction product absorbance measurement, 460 nm) in the presence of organic solvent	Dimethyl Sulfoxide (DMSO)	40%	0.11	14.17	5.1	mM	100 mM glycine buffer	8.6	37°C	1,2-Dibromoethane	2-Bromoethanol, Bromide ion	—	—	Classical aqueous control (in mM)
T148L/G171V/A172VC176F	Activity - Michaelis-Menten	Km (mM) measured by absorbance spectrophotometry (colorimetric assay, Br ion, mercuric thiocyanate and ferric ammonium sulfate reaction product absorbance measurement, 460 nm) in the presence of organic solvent	Dimethyl Sulfoxide (DMSO)	40%	0.11	14.17	1.4	mM	100 mM glycine buffer	8.6	37°C	1,2-Dibromoethane	2-Bromoethanol, Bromide ion	—	—	Classical aqueous control (in mM)
T148L/G171Q/A172V/C176F	Activity - Michaelis-Menten	Km (mM) measured by absorbance spectrophotometry (colorimetric assay, Br ion, mercuric thiocyanate and ferric ammonium sulfate reaction product absorbance measurement, 460 nm) in the presence of organic solvent	Dimethyl Sulfoxide (DMSO)	40%	0.13	14.17	0.88	mM	100 mM glycine buffer	8.6	37°C	1,2-Dibromoethane	2-Bromoethanol, Bromide ion	—	—	Classical aqueous control (in mM) \| note actually Hill coefficient for the DMSO (Dimethyl Sulfoxide) measurement with n=2.43 here approximated to be comparable to the Km
D78G/F80S/T148L/G171Q/A172V/C176F/N227T/W240Y/P291A/A292G	Activity - Michaelis-Menten	Km (mM) measured by absorbance spectrophotometry (colorimetric assay, Br ion, mercuric thiocyanate and ferric ammonium sulfate reaction product absorbance measurement, 460 nm) in the presence of organic solvent	Dimethyl Sulfoxide (DMSO)	40%	1.7	14.17	1.08	mM	100 mM glycine buffer	8.6	37°C	1,2-Dibromoethane	2-Bromoethanol, Bromide ion	—	—	Classical aqueous control (in mM) \| note actually Hill coefficient for the DMSO (Dimethyl Sulfoxide) measurement with n=2.43 here approximated to be comparable to the Km
A172V	Activity - Michaelis-Menten	Km (mM) measured by absorbance spectrophotometry (colorimetric assay, Br ion, mercuric thiocyanate and ferric ammonium sulfate reaction product absorbance measurement, 460 nm) in the presence of organic solvent	Dimethyl Sulfoxide (DMSO)	40%	1.17	14.17	5.47	mM	100 mM glycine buffer	8.6	37°C	1,2-Dibromoethane	2-Bromoethanol, Bromide ion	—	—	Classical aqueous control (in mM)
L95V/A172V	Activity - Michaelis-Menten	Km (mM) measured by absorbance spectrophotometry (colorimetric assay, Br ion, mercuric thiocyanate and ferric ammonium sulfate reaction product absorbance measurement, 460 nm) in the presence of organic solvent	Dimethyl Sulfoxide (DMSO)	40%	0.85	14.17	4.09	mM	100 mM glycine buffer	8.6	37°C	1,2-Dibromoethane	2-Bromoethanol, Bromide ion	—	—	Classical aqueous control (in mM)
L95V/A172V	Stability - C50	Volume percentage of DMSO (Dimethyl Sulfoxide) for which half enzyme is unfolded at 23°C, measured by fluorescence spectrophotometry	Dimethyl Sulfoxide (DMSO)	—	—	49.0	53.3	%	50 mM phosphate buffer	7.5	23°C	—	—	—	—	Classical aqueous control (in %)
L95V	Stability - C50	Volume percentage of DMSO (Dimethyl Sulfoxide) for which half enzyme is unfolded at 23°C, measured by fluorescence spectrophotometry	Dimethyl Sulfoxide (DMSO)	—	—	49.0	48.8	%	50 mM phosphate buffer	7.5	23°C	—	—	—	—	Classical aqueous control (in %)
A172V	Stability - C50	Volume percentage of DMSO (Dimethyl Sulfoxide) for which half enzyme is unfolded at 23°C, measured by fluorescence spectrophotometry	Dimethyl Sulfoxide (DMSO)	—	—	49.0	48.5	%	50 mM phosphate buffer	7.5	23°C	—	—	—	—	Classical aqueous control (in %)
D78G/F80S/T148L/G171Q/A172V/C176F/N227T/W240Y/P291A/A292G	Stability - C50	Volume percentage of DMSO (Dimethyl Sulfoxide) for which half enzyme is unfolded at 23°C, measured by fluorescence spectrophotometry	Dimethyl Sulfoxide (DMSO)	—	—	49.0	56.2	%	50 mM phosphate buffer	7.5	23°C	—	—	—	—	Classical aqueous control (in %)
T148L/G171Q/A172V/C176F	Stability - C50	Volume percentage of DMSO (Dimethyl Sulfoxide) for which half enzyme is unfolded at 23°C, measured by fluorescence spectrophotometry	Dimethyl Sulfoxide (DMSO)	—	—	49.0	60.8	%	50 mM phosphate buffer	7.5	23°C	—	—	—	—	Classical aqueous control (in %)
D78G/F80S/N227T/W240Y/P291A/A292G	Stability - C50	Volume percentage of DMSO (Dimethyl Sulfoxide) for which half enzyme is unfolded at 23°C, measured by fluorescence spectrophotometry	Dimethyl Sulfoxide (DMSO)	—	—	49.0	39.1	%	50 mM phosphate buffer	7.5	23°C	—	—	—	—	Classical aqueous control (in %)
T148L/G171V/A172V/C176F	Stability - C50	Volume percentage of DMSO (Dimethyl Sulfoxide) for which half enzyme is unfolded at 23°C, measured by fluorescence spectrophotometry	Dimethyl Sulfoxide (DMSO)	—	—	49.0	63.0	%	50 mM phosphate buffer	7.5	23°C	—	—	—	—	Classical aqueous control (in %)
T148L/G171L/A172V/C176F	Stability - C50	Volume percentage of DMSO (Dimethyl Sulfoxide) for which half enzyme is unfolded at 23°C, measured by fluorescence spectrophotometry	Dimethyl Sulfoxide (DMSO)	—	—	49.0	63.1	%	50 mM phosphate buffer	7.5	23°C	—	—	—	—	Classical aqueous control (in %)
L95V/A172V	Stability - C50	Volume percentage of DMSO (Dimethyl Sulfoxide) for which half enzyme is unfolded after 30 minutes incubation at 37°C in the presence of organic solvent, measured by fluorescence spectrophotometry	Dimethyl Sulfoxide (DMSO)	—	—	29.1	43.0	%	50 mM phosphate buffer	7.5	37°C	—	—	—	—	Classical aqueous control (in %)
L95V	Stability - C50	Volume percentage of DMSO (Dimethyl Sulfoxide) for which half enzyme is unfolded after 30 minutes incubation at 37°C in the presence of organic solvent, measured by fluorescence spectrophotometry	Dimethyl Sulfoxide (DMSO)	—	—	29.1	31.9	%	50 mM phosphate buffer	7.5	37°C	—	—	—	—	Classical aqueous control (in %)
A172V	Stability - C50	Volume percentage of DMSO (Dimethyl Sulfoxide) for which half enzyme is unfolded after 30 minutes incubation at 37°C in the presence of organic solvent, measured by fluorescence spectrophotometry	Dimethyl Sulfoxide (DMSO)	—	—	29.1	42.0	%	50 mM phosphate buffer	7.5	37°C	—	—	—	—	Classical aqueous control (in %)
D78G/F80S/T148L/G171Q/A172V/C176F/N227T/W240Y/P291A/A292G	Stability - C50	Volume percentage of DMSO (Dimethyl Sulfoxide) for which half enzyme is unfolded after 30 minutes incubation at 37°C in the presence of organic solvent, measured by fluorescence spectrophotometry	Dimethyl Sulfoxide (DMSO)	—	—	29.1	51.7	%	50 mM phosphate buffer	7.5	37°C	—	—	—	—	Classical aqueous control (in %)
T148L/G171Q/A172V/C176F	Stability - C50	Volume percentage of DMSO (Dimethyl Sulfoxide) for which half enzyme is unfolded after 30 minutes incubation at 37°C in the presence of organic solvent, measured by fluorescence spectrophotometry	Dimethyl Sulfoxide (DMSO)	—	—	29.1	54.9	%	50 mM phosphate buffer	7.5	37°C	—	—	—	—	Classical aqueous control (in %)
D78G/F80S/N227T/W240Y/P291A/A292G	Stability - C50	Volume percentage of DMSO (Dimethyl Sulfoxide) for which half enzyme is unfolded after 30 minutes incubation at 37°C in the presence of organic solvent, measured by fluorescence spectrophotometry	Dimethyl Sulfoxide (DMSO)	—	—	29.1	31.3	%	50 mM phosphate buffer	7.5	37°C	—	—	—	—	Classical aqueous control (in %)
T148L/G171V/A172V/C176F	Stability - C50	Volume percentage of DMSO (Dimethyl Sulfoxide) for which half enzyme is unfolded after 30 minutes incubation at 37°C in the presence of organic solvent, measured by fluorescence spectrophotometry	Dimethyl Sulfoxide (DMSO)	—	—	29.1	56.1	%	50 mM phosphate buffer	7.5	37°C	—	—	—	—	Classical aqueous control (in %)
T148L/G171L/A172V/C176F	Stability - C50	Volume percentage of DMSO (Dimethyl Sulfoxide) for which half enzyme is unfolded after 30 minutes incubation at 37°C in the presence of organic solvent, measured by fluorescence spectrophotometry	Dimethyl Sulfoxide (DMSO)	—	—	29.1	58.1	%	50 mM phosphate buffer	7.5	37°C	—	—	—	—	Classical aqueous control (in %)
T148L/G171V/A172VC176F	Stability - Inactivation Rate Constant	Ksi (mM) measured by absorbance spectrophotometry (colorimetric assay, Br ion, mercuric thiocyanate and ferric ammonium sulfate reaction product absorbance measurement, 460 nm) in the presence of organic solvent	Dimethyl Sulfoxide (DMSO)	40%	0.42	0.0	6.69	mM	100 mM glycine buffer	8.6	37°C	1,2-Dibromoethane	2-Bromoethanol, Bromide ion	—	—	Classical aqueous control (in mM)
T148L/G171Q/A172V/C176F/	Stability - Inactivation Rate Constant	Ksi (mM) measured by absorbance spectrophotometry (colorimetric assay, Br ion, mercuric thiocyanate and ferric ammonium sulfate reaction product absorbance measurement, 460 nm) in the presence of organic solvent	Dimethyl Sulfoxide (DMSO)	40%	0.41	0.0	13.14	mM	100 mM glycine buffer	8.6	37°C	1,2-Dibromoethane	2-Bromoethanol, Bromide ion	—	—	Classical aqueous control (in mM)
T148L/G171L/A172VC176F	Stability - Inactivation Rate Constant	Ksi (mM) measured by absorbance spectrophotometry (colorimetric assay, Br ion, mercuric thiocyanate and ferric ammonium sulfate reaction product absorbance measurement, 460 nm) in the presence of organic solvent	Dimethyl Sulfoxide (DMSO)	40%	0.48	0.0	2.34	mM	100 mM glycine buffer	8.6	37°C	1,2-Dibromoethane	2-Bromoethanol, Bromide ion	—	—	Classical aqueous control (in mM)
D78G/F80S/T148L/G171Q/A172V/C176F/N227T/W240Y/P291A/A292G	Stability - Inactivation Rate Constant	Ksi (mM) measured by absorbance spectrophotometry (colorimetric assay, Br ion, mercuric thiocyanate and ferric ammonium sulfate reaction product absorbance measurement, 460 nm) in the presence of organic solvent	Dimethyl Sulfoxide (DMSO)	40%	0.24	0.0	41.44	mM	100 mM glycine buffer	8.6	37°C	1,2-Dibromoethane	2-Bromoethanol, Bromide ion	—	—	Classical aqueous control (in mM)
A172V	Stability - Inactivation Rate Constant	Ksi (mM) measured by absorbance spectrophotometry (colorimetric assay, Br ion, mercuric thiocyanate and ferric ammonium sulfate reaction product absorbance measurement, 460 nm) in the presence of organic solvent	Dimethyl Sulfoxide (DMSO)	40%	3.45	0.0	0.0	mM	100 mM glycine buffer	8.6	37°C	1,2-Dibromoethane	2-Bromoethanol, Bromide ion	—	—	Classical aqueous control (in mM)
L95V/A172V	Stability - Inactivation Rate Constant	Ksi (mM) measured by absorbance spectrophotometry (colorimetric assay, Br ion, mercuric thiocyanate and ferric ammonium sulfate reaction product absorbance measurement, 460 nm) in the presence of organic solvent	Dimethyl Sulfoxide (DMSO)	40%	6.55	0.0	0.0	mM	100 mM glycine buffer	8.6	37°C	1,2-Dibromoethane	2-Bromoethanol, Bromide ion	—	—	Classical aqueous control (in mM)
A172V	Stability - Tm	Thermal stability measured by DSC in the presence of organic solvent	Acetonitrile	10%	56.2	36.8	44.4	°C	50 mM phosphate buffer	7.5	—	—	—	—	—	Classical aqueous control (in °C)
T148L/G171Q/A172V/C176F	Stability - Tm	Thermal stability measured by DSC in the presence of organic solvent	tert-Butanol	20%	68.2	30.6	42.2	°C	50 mM phosphate buffer	7.5	—	—	—	—	—	Classical aqueous control (in °C)
T148L/G171Q/A172V/C176F	Stability - Tm	Thermal stability measured by DSC in the presence of organic solvent	Dimethyl Sulfoxide (DMSO)	40%	68.2	39.0	60.4	°C	50 mM phosphate buffer	7.5	—	—	—	—	—	Classical aqueous control (in °C)
T148L/G171Q/A172V/C176F	Stability - Tm	Thermal stability measured by DSC in the presence of organic solvent	Acetonitrile	10%	68.2	36.8	56.5	°C	50 mM phosphate buffer	7.5	—	—	—	—	—	Classical aqueous control (in °C)
A172V	Stability - Tm	Thermal stability measured by DSC in the presence of organic solvent	Dimethyl Sulfoxide (DMSO)	40%	56.2	39.0	48.3	°C	50 mM phosphate buffer	7.5	—	—	—	—	—	Classical aqueous control (in °C)
G171Q/A172V/C176F	Stability - Tm	Thermal stability measured by DSC in the presence of organic solvent	tert-Butanol	20%	65.9	30.6	42.1	°C	50 mM phosphate buffer	7.5	—	—	—	—	—	Classical aqueous control (in °C)
G171Q/A172V/C176F	Stability - Tm	Thermal stability measured by DSC in the presence of organic solvent	Dimethyl Sulfoxide (DMSO)	40%	65.9	39.0	57.3	°C	50 mM phosphate buffer	7.5	—	—	—	—	—	Classical aqueous control (in °C)
G171Q/A172V/C176F	Stability - Tm	Thermal stability measured by DSC in the presence of organic solvent	Acetonitrile	10%	65.9	36.8	52.0	°C	50 mM phosphate buffer	7.5	—	—	—	—	—	Classical aqueous control (in °C)
A172V/C176F	Stability - Tm	Thermal stability measured by DSC in the presence of organic solvent	tert-Butanol	20%	63.8	30.6	41.8	°C	50 mM phosphate buffer	7.5	—	—	—	—	—	Classical aqueous control (in °C)
A172V/C176F	Stability - Tm	Thermal stability measured by DSC in the presence of organic solvent	Dimethyl Sulfoxide (DMSO)	40%	63.8	39.0	54.8	°C	50 mM phosphate buffer	7.5	—	—	—	—	—	Classical aqueous control (in °C)
A172V/C176F	Stability - Tm	Thermal stability measured by DSC in the presence of organic solvent	Acetonitrile	10%	63.8	36.8	50.5	°C	50 mM phosphate buffer	7.5	—	—	—	—	—	Classical aqueous control (in °C)
A172V	Stability - Tm	Thermal stability measured by DSC in the presence of organic solvent	tert-Butanol	20%	56.2	30.6	36.9	°C	50 mM phosphate buffer	7.5	—	—	—	—	—	Classical aqueous control (in °C)

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Mutations in this dataset (17) — Tana Koudelakova et al. (2013)

WT D73G/E98V D78G/F80S/N227T/W240Y/P291A/A292G D78G/F80S/T148L/G171Q/A172V/C176F/N227T/W240Y/P291A/A292G L95V L95V/A172V F144I/C176G T148L/G171L/A172V/C176F T148L/G171L/A172VC176F T148L/G171Q/A172V/C176F T148L/G171Q/A172V/C176F/ T148L/G171V/A172V/C176F T148L/G171V/A172VC176F G171Q/A172V/C176F A172V A172V/C176F V245L