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 6 of 7

Property	Assay	Solvent	Solvent Volume	Aqueous Reference	Measured Value	Units	Extraction method	Data type	Solution	pH	Temperature	Substrate(s)	Product(s)	Cofactor(s)	Shaking	Comments
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 % v/v	100	4	%	Direct	Continuous	glycine 100 mM	8.6	37°C	? 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 % v/v	100	125	%	Direct	Continuous	glycine 100 mM	8.6	37°C	? 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 % v/v	100	0	%	Direct	Continuous	glycine 100 mM	8.6	37°C	? 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 % v/v	100	0	%	Direct	Continuous	glycine 100 mM	8.6	37°C	? 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 % v/v	100	0	%	Direct	Continuous	glycine 100 mM	8.6	37°C	? 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 % v/v	100	0	%	Direct	Continuous	glycine 100 mM	8.6	37°C	? 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 % v/v	100	0	%	Direct	Continuous	glycine 100 mM	8.6	37°C	? 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 % v/v	100	0	%	Direct	Continuous	glycine 100 mM	8.6	37°C	? 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 % v/v	100	0	%	Direct	Continuous	glycine 100 mM	8.6	37°C	? 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 % v/v	100	0	%	Direct	Continuous	glycine 100 mM	8.6	37°C	? 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 % v/v	100	0	%	Direct	Continuous	glycine 100 mM	8.6	37°C	? 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 % v/v	100	0	%	Direct	Continuous	glycine 100 mM	8.6	37°C	? 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)	Direct	Continuous	glycine 100 mM	8.6	37°C	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)	Direct	Continuous	glycine 100 mM	8.6	37°C	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)	Direct	Continuous	glycine 100 mM	8.6	37°C	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)	Direct	Continuous	glycine 100 mM	8.6	37°C	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)	Direct	Continuous	glycine 100 mM	8.6	37°C	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)	Direct	Continuous	glycine 100 mM	8.6	37°C	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)	Direct	Continuous	glycine 100 mM	8.6	37°C	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)	Direct	Continuous	glycine 100 mM	8.6	37°C	1-Iodohexane	1-Hexanol , Iodide ion	—	—	Not applicable control (in % (v/v))

—

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 % v/v

100

112

Direct

Continuous

glycine 100 mM

8.6

37°C

? mM 1-Iodohexane

1-Hexanol, Iodide ion

—

Classical aqueous control (in %)

—

Activity - Classical

Formamide

5 % v/v

100

150

Direct

Continuous

glycine 100 mM

8.6

37°C

? mM 1-Iodohexane

1-Hexanol, Iodide ion

—

Classical aqueous control (in %)

—

Activity - Classical

Ethylene Glycol

5 % v/v

100

113

Direct

Continuous

glycine 100 mM

8.6

37°C

? mM 1-Iodohexane

1-Hexanol, Iodide ion

—

Classical aqueous control (in %)

—

Activity - Classical

PEG-1000

5 % v/v

100

Direct

Continuous

glycine 100 mM

8.6

37°C

? mM 1-Iodohexane

1-Hexanol, Iodide ion

—

Classical aqueous control (in %)

—

Activity - Classical

PEG-10000

5 % v/v

100

Direct

Continuous

glycine 100 mM

8.6

37°C

? mM 1-Iodohexane

1-Hexanol, Iodide ion

—

Classical aqueous control (in %)

—

Activity - Classical

Dimethylformamide (DMF)

5 % v/v

100

Direct

Continuous

glycine 100 mM

8.6

37°C

? mM 1-Iodohexane

1-Hexanol, Iodide ion

—

Classical aqueous control (in %)

—

Activity - Classical

Dimethyl Sulfoxide (DMSO)

5 % v/v

100

111

Direct

Continuous

glycine 100 mM

8.6

37°C

? mM 1-Iodohexane

1-Hexanol, Iodide ion

—

Classical aqueous control (in %)

—

Activity - Classical

Methanol

5 % v/v

100

150

Direct

Continuous

glycine 100 mM

8.6

37°C

? mM 1-Iodohexane

1-Hexanol, Iodide ion

—

Classical aqueous control (in %)

—

Activity - Classical

1,4-Dioxane

5 % v/v

100

Direct

Continuous

glycine 100 mM

8.6

37°C

? mM 1-Iodohexane

1-Hexanol, Iodide ion

—

Classical aqueous control (in %)

—

Activity - Classical

Acetonitrile

5 % v/v

100

Direct

Continuous

glycine 100 mM

8.6

37°C

? mM 1-Iodohexane

1-Hexanol, Iodide ion

—

Classical aqueous control (in %)

—

Activity - Classical

Ethanol

5 % v/v

100

120

Direct

Continuous

glycine 100 mM

8.6

37°C

? mM 1-Iodohexane

1-Hexanol, Iodide ion

—

Classical aqueous control (in %)

—

Activity - Classical

Acetone

5 % v/v

100

108

Direct

Continuous

glycine 100 mM

8.6

37°C

? mM 1-Iodohexane

1-Hexanol, Iodide ion

—

Classical aqueous control (in %)

—

Activity - Classical

Isopropanol

5 % v/v

100

111

Direct

Continuous

glycine 100 mM

8.6

37°C

? mM 1-Iodohexane

1-Hexanol, Iodide ion

—

Classical aqueous control (in %)

—

Activity - Classical

Tetrahydrofuran (THF)

5 % v/v

100

Direct

Continuous

glycine 100 mM

8.6

37°C

? mM 1-Iodohexane

1-Hexanol, Iodide ion

—

Classical aqueous control (in %)

—

Activity - Classical

Glycerol

10 % v/v

100

120

Direct

Continuous

glycine 100 mM

8.6

37°C

? mM 1-Iodohexane

1-Hexanol, Iodide ion

—

Classical aqueous control (in %)

—

Activity - Classical

Formamide

10 % v/v

100

112

Direct

Continuous

glycine 100 mM

8.6

37°C

? mM 1-Iodohexane

1-Hexanol, Iodide ion

—

Classical aqueous control (in %)

—

Activity - Classical

Ethylene Glycol

10 % v/v

100

110

Direct

Continuous

glycine 100 mM

8.6

37°C

? mM 1-Iodohexane

1-Hexanol, Iodide ion

—

Classical aqueous control (in %)

—

Activity - Classical

PEG-1000

10 % v/v

100

Direct

Continuous

glycine 100 mM

8.6

37°C

? mM 1-Iodohexane

1-Hexanol, Iodide ion

—

Classical aqueous control (in %)

—

Activity - Classical

PEG-10000

10 % v/v

100

Direct

Continuous

glycine 100 mM

8.6

37°C

? mM 1-Iodohexane

1-Hexanol, Iodide ion

—

Classical aqueous control (in %)

—

Activity - Classical

Dimethylformamide (DMF)

10 % v/v

100

Direct

Continuous

glycine 100 mM

8.6

37°C

? mM 1-Iodohexane

1-Hexanol, Iodide ion

—

Classical aqueous control (in %)

—

Activity - Classical

Dimethyl Sulfoxide (DMSO)

10 % v/v

100

113

Direct

Continuous

glycine 100 mM

8.6

37°C

? mM 1-Iodohexane

1-Hexanol, Iodide ion

—

Classical aqueous control (in %)

—

Activity - Classical

Methanol

10 % v/v

100

141

Direct

Continuous

glycine 100 mM

8.6

37°C

? mM 1-Iodohexane

1-Hexanol, Iodide ion

—

Classical aqueous control (in %)

—

Activity - Classical

1,4-Dioxane

10 % v/v

100

Direct

Continuous

glycine 100 mM

8.6

37°C

? mM 1-Iodohexane

1-Hexanol, Iodide ion

—

Classical aqueous control (in %)

—

Activity - Classical

Acetonitrile

10 % v/v

100

Direct

Continuous

glycine 100 mM

8.6

37°C

? mM 1-Iodohexane

1-Hexanol, Iodide ion

—

Classical aqueous control (in %)

—

Activity - Classical

Ethanol

10 % v/v

100

117

Direct

Continuous

glycine 100 mM

8.6

37°C

? mM 1-Iodohexane

1-Hexanol, Iodide ion

—

Classical aqueous control (in %)

—

Activity - Classical

Acetone

10 % v/v

100

Direct

Continuous

glycine 100 mM

8.6

37°C

? mM 1-Iodohexane

1-Hexanol, Iodide ion

—

Classical aqueous control (in %)

—

Activity - Classical

Isopropanol

10 % v/v

100

Direct

Continuous

glycine 100 mM

8.6

37°C

? mM 1-Iodohexane

1-Hexanol, Iodide ion

—

Classical aqueous control (in %)

—

Activity - Classical

Tetrahydrofuran (THF)

10 % v/v

100

Direct

Continuous

glycine 100 mM

8.6

37°C

? mM 1-Iodohexane

1-Hexanol, Iodide ion

—

Classical aqueous control (in %)

—

Activity - Classical

Glycerol

20 % v/v

100

125

Direct

Continuous

glycine 100 mM

8.6

37°C

? mM 1-Iodohexane

1-Hexanol, Iodide ion

—

Classical aqueous control (in %)

—

Activity - Classical

Formamide

20 % v/v

100

Direct

Continuous

glycine 100 mM

8.6

37°C

? mM 1-Iodohexane

1-Hexanol, Iodide ion

—

Classical aqueous control (in %)

—

Activity - Classical

Ethylene Glycol

20 % v/v

100

111

Direct

Continuous

glycine 100 mM

8.6

37°C

? mM 1-Iodohexane

1-Hexanol, Iodide ion

—

Classical aqueous control (in %)

—

Activity - Classical

PEG-1000

20 % v/v

100

Direct

Continuous

glycine 100 mM

8.6

37°C

? mM 1-Iodohexane

1-Hexanol, Iodide ion

—

Classical aqueous control (in %)

—

Activity - Classical

PEG-10000

20 % v/v

100

Direct

Continuous

glycine 100 mM

8.6

37°C

? mM 1-Iodohexane

1-Hexanol, Iodide ion

—

Classical aqueous control (in %)

—

Activity - Classical

Dimethylformamide (DMF)

20 % v/v

100

Direct

Continuous

glycine 100 mM

8.6

37°C

? mM 1-Iodohexane

1-Hexanol, Iodide ion

—

Classical aqueous control (in %)

—

Activity - Classical

Dimethyl Sulfoxide (DMSO)

20 % v/v

100

107

Direct

Continuous

glycine 100 mM

8.6

37°C

? mM 1-Iodohexane

1-Hexanol, Iodide ion

—

Classical aqueous control (in %)

—

Activity - Classical

Methanol

20 % v/v

100

Direct

Continuous

glycine 100 mM

8.6

37°C

? mM 1-Iodohexane

1-Hexanol, Iodide ion

—

Classical aqueous control (in %)

—

Activity - Classical

1,4-Dioxane

20 % v/v

100

Direct

Continuous

glycine 100 mM

8.6

37°C

? mM 1-Iodohexane

1-Hexanol, Iodide ion

—

Classical aqueous control (in %)

—

Activity - Classical

Acetonitrile

20 % v/v

100

Direct

Continuous

glycine 100 mM

8.6

37°C

? mM 1-Iodohexane

1-Hexanol, Iodide ion

—

Classical aqueous control (in %)

—

Activity - Classical

Ethanol

20 % v/v

100

Direct

Continuous

glycine 100 mM

8.6

37°C

? mM 1-Iodohexane

1-Hexanol, Iodide ion

—

Classical aqueous control (in %)

—

Activity - Classical

Acetone

20 % v/v

100

Direct

Continuous

glycine 100 mM

8.6

37°C

? mM 1-Iodohexane

1-Hexanol, Iodide ion

—

Classical aqueous control (in %)

—

Activity - Classical

Isopropanol

20 % v/v

100

Direct

Continuous

glycine 100 mM

8.6

37°C

? mM 1-Iodohexane

1-Hexanol, Iodide ion

—

Classical aqueous control (in %)

—

Activity - Classical

Tetrahydrofuran (THF)

20 % v/v

100

Direct

Continuous

glycine 100 mM

8.6

37°C

? mM 1-Iodohexane

1-Hexanol, Iodide ion

—

Classical aqueous control (in %)

—

Activity - Classical

Glycerol

30 % v/v

100

107

Direct

Continuous

glycine 100 mM

8.6

37°C

? mM 1-Iodohexane

1-Hexanol, Iodide ion

—

Classical aqueous control (in %)

—

Activity - Classical

Formamide

30 % v/v

100

Direct

Continuous

glycine 100 mM

8.6

37°C

? mM 1-Iodohexane

1-Hexanol, Iodide ion

—

Classical aqueous control (in %)

—

Activity - Classical

Ethylene Glycol

30 % v/v

100

121

Direct

Continuous

glycine 100 mM

8.6

37°C

? mM 1-Iodohexane

1-Hexanol, Iodide ion

—

Classical aqueous control (in %)

—

Activity - Classical

PEG-1000

30 % v/v

100

108

Direct

Continuous

glycine 100 mM

8.6

37°C

? mM 1-Iodohexane

1-Hexanol, Iodide ion

—

Classical aqueous control (in %)

—

Activity - Classical

PEG-10000

30 % v/v

100

Direct

Continuous

glycine 100 mM

8.6

37°C

? mM 1-Iodohexane

1-Hexanol, Iodide ion

—

Classical aqueous control (in %)

—

Activity - Classical

Dimethylformamide (DMF)

30 % v/v

100

Direct

Continuous

glycine 100 mM

8.6

37°C

? mM 1-Iodohexane

1-Hexanol, Iodide ion

—

Classical aqueous control (in %)

—

Activity - Classical

Dimethyl Sulfoxide (DMSO)

30 % v/v

100

Direct

Continuous

glycine 100 mM

8.6

37°C

? mM 1-Iodohexane

1-Hexanol, Iodide ion

—

Classical aqueous control (in %)

—

Activity - Classical

Methanol

30 % v/v

100

Direct

Continuous

glycine 100 mM

8.6

37°C

? mM 1-Iodohexane

1-Hexanol, Iodide ion

—

Classical aqueous control (in %)

—

Activity - Classical

1,4-Dioxane

30 % v/v

100

Direct

Continuous

glycine 100 mM

8.6

37°C

? mM 1-Iodohexane

1-Hexanol, Iodide ion

—

Classical aqueous control (in %)

—

Activity - Classical

Acetonitrile

30 % v/v

100

Direct

Continuous

glycine 100 mM

8.6

37°C

? mM 1-Iodohexane

1-Hexanol, Iodide ion

—

Classical aqueous control (in %)

—

Activity - Classical

Ethanol

30 % v/v

100

Direct

Continuous

glycine 100 mM

8.6

37°C

? mM 1-Iodohexane

1-Hexanol, Iodide ion

—

Classical aqueous control (in %)

—

Activity - Classical

Acetone

30 % v/v

100

Direct

Continuous

glycine 100 mM

8.6

37°C

? mM 1-Iodohexane

1-Hexanol, Iodide ion

—

Classical aqueous control (in %)

—

Activity - Classical

Isopropanol

30 % v/v

100

Direct

Continuous

glycine 100 mM

8.6

37°C

? mM 1-Iodohexane

1-Hexanol, Iodide ion

—

Classical aqueous control (in %)

—

Activity - Classical

Tetrahydrofuran (THF)

30 % v/v

100

Direct

Continuous

glycine 100 mM

8.6

37°C

? mM 1-Iodohexane

1-Hexanol, Iodide ion

—

Classical aqueous control (in %)

—

Activity - Classical

Glycerol

40 % v/v

100

110

Direct

Continuous

glycine 100 mM

8.6

37°C

? mM 1-Iodohexane

1-Hexanol, Iodide ion

—

Classical aqueous control (in %)

—

Activity - Classical

Formamide

40 % v/v

100

Direct

Continuous

glycine 100 mM

8.6

37°C

? mM 1-Iodohexane

1-Hexanol, Iodide ion

—

Classical aqueous control (in %)

—

Activity - Classical

Ethylene Glycol

40 % v/v

100

126

Direct

Continuous

glycine 100 mM

8.6

37°C

? mM 1-Iodohexane

1-Hexanol, Iodide ion

—

Classical aqueous control (in %)

—

Activity - Classical

PEG-1000

40 % v/v

100

117

Direct

Continuous

glycine 100 mM

8.6

37°C

? mM 1-Iodohexane

1-Hexanol, Iodide ion

—

Classical aqueous control (in %)

—

Activity - Classical

PEG-10000

40 % v/v

100

Direct

Continuous

glycine 100 mM

8.6

37°C

? mM 1-Iodohexane

1-Hexanol, Iodide ion

—

Classical aqueous control (in %)

—

Activity - Classical

Dimethylformamide (DMF)

40 % v/v

100

Direct

Continuous

glycine 100 mM

8.6

37°C

? mM 1-Iodohexane

1-Hexanol, Iodide ion

—

Classical aqueous control (in %)

—

Activity - Classical

Dimethyl Sulfoxide (DMSO)

40 % v/v

100

Direct

Continuous

glycine 100 mM

8.6

37°C

? mM 1-Iodohexane

1-Hexanol, Iodide ion

—

Classical aqueous control (in %)

—

Activity - Classical

Methanol

40 % v/v

100

Direct

Continuous

glycine 100 mM

8.6

37°C

? mM 1-Iodohexane

1-Hexanol, Iodide ion

—

Classical aqueous control (in %)

—

Activity - Classical

1,4-Dioxane

40 % v/v

100

Direct

Continuous

glycine 100 mM

8.6

37°C

? mM 1-Iodohexane

1-Hexanol, Iodide ion

—

Classical aqueous control (in %)

—

Activity - Classical

Acetonitrile

40 % v/v

100

Direct

Continuous

glycine 100 mM

8.6

37°C

? mM 1-Iodohexane

1-Hexanol, Iodide ion

—

Classical aqueous control (in %)

—

Activity - Classical

Ethanol

40 % v/v

100

Direct

Continuous

glycine 100 mM

8.6

37°C

? mM 1-Iodohexane

1-Hexanol, Iodide ion

—

Classical aqueous control (in %)

—

Activity - Classical

Acetone

40 % v/v

100

Direct

Continuous

glycine 100 mM

8.6

37°C

? mM 1-Iodohexane

1-Hexanol, Iodide ion

—

Classical aqueous control (in %)

—

Activity - Classical

Isopropanol

40 % v/v

100

Direct

Continuous

glycine 100 mM

8.6

37°C

? mM 1-Iodohexane

1-Hexanol, Iodide ion

—

Classical aqueous control (in %)

—

Activity - Classical

Tetrahydrofuran (THF)

40 % v/v

100

Direct

Continuous

glycine 100 mM

8.6

37°C

? mM 1-Iodohexane

1-Hexanol, Iodide ion

—

Classical aqueous control (in %)

—

Activity - Classical

Glycerol

50 % v/v

100

Direct

Continuous

glycine 100 mM

8.6

37°C

? mM 1-Iodohexane

1-Hexanol, Iodide ion

—

Classical aqueous control (in %)

—

Activity - Classical

Formamide

50 % v/v

100

Direct

Continuous

glycine 100 mM

8.6

37°C

? mM 1-Iodohexane

1-Hexanol, Iodide ion

—

Classical aqueous control (in %)

—

Activity - Classical

Ethylene Glycol

50 % v/v

100

Direct

Continuous

glycine 100 mM

8.6

37°C

? mM 1-Iodohexane

1-Hexanol, Iodide ion

—

Classical aqueous control (in %)

—

Activity - Classical

PEG-1000

50 % v/v

100

119

Direct

Continuous

glycine 100 mM

8.6

37°C

? mM 1-Iodohexane

1-Hexanol, Iodide ion

—

Classical aqueous control (in %)

—

Activity - Classical

PEG-10000

50 % v/v

100

Direct

Continuous

glycine 100 mM

8.6

37°C

? mM 1-Iodohexane

1-Hexanol, Iodide ion

—

Classical aqueous control (in %)

—

Activity - Classical

Dimethylformamide (DMF)

50 % v/v

100

Direct

Continuous

glycine 100 mM

8.6

37°C

? mM 1-Iodohexane

1-Hexanol, Iodide ion

—

Classical aqueous control (in %)

—

Activity - Classical

Dimethyl Sulfoxide (DMSO)

50 % v/v

100

Direct

Continuous

glycine 100 mM

8.6

37°C

? mM 1-Iodohexane

1-Hexanol, Iodide ion

—

Classical aqueous control (in %)

—

Activity - Classical

Methanol

50 % v/v

100

Direct

Continuous

glycine 100 mM

8.6

37°C

? mM 1-Iodohexane

1-Hexanol, Iodide ion

—

Classical aqueous control (in %)

—

Activity - Classical

1,4-Dioxane

50 % v/v

100

Direct

Continuous

glycine 100 mM

8.6

37°C

? mM 1-Iodohexane

1-Hexanol, Iodide ion

—

Classical aqueous control (in %)

—

Activity - Classical

Acetonitrile

50 % v/v

100

Direct

Continuous

glycine 100 mM

8.6

37°C

? mM 1-Iodohexane

1-Hexanol, Iodide ion

—

Classical aqueous control (in %)

—

Activity - Classical

Ethanol

50 % v/v

100

Direct

Continuous

glycine 100 mM

8.6

37°C

? mM 1-Iodohexane

1-Hexanol, Iodide ion

—

Classical aqueous control (in %)

—

Activity - Classical

Acetone

50 % v/v

100

Direct

Continuous

glycine 100 mM

8.6

37°C

? mM 1-Iodohexane

1-Hexanol, Iodide ion

—

Classical aqueous control (in %)

—

Activity - Classical

Isopropanol

50 % v/v

100

Direct

Continuous

glycine 100 mM

8.6

37°C

? mM 1-Iodohexane

1-Hexanol, Iodide ion

—

Classical aqueous control (in %)

—

Activity - Classical

Tetrahydrofuran (THF)

50 % v/v

100

Direct

Continuous

glycine 100 mM

8.6

37°C

? mM 1-Iodohexane

1-Hexanol, Iodide ion

—

Classical aqueous control (in %)

—

Activity - Classical

Glycerol

60 % v/v

100

Direct

Continuous

glycine 100 mM

8.6

37°C

? mM 1-Iodohexane

1-Hexanol, Iodide ion

—

Classical aqueous control (in %)

—

Activity - Classical

Formamide

60 % v/v

100

Direct

Continuous

glycine 100 mM

8.6

37°C

? mM 1-Iodohexane

1-Hexanol, Iodide ion

—

Classical aqueous control (in %)

—

Activity - Classical

Ethylene Glycol

60 % v/v

100

Direct

Continuous

glycine 100 mM

8.6

37°C

? mM 1-Iodohexane

1-Hexanol, Iodide ion

—

Classical aqueous control (in %)

—

Activity - Classical

PEG-1000

60 % v/v

100

123

Direct

Continuous

glycine 100 mM

8.6

37°C

? mM 1-Iodohexane

1-Hexanol, Iodide ion

—

Classical aqueous control (in %)

—

Activity - Classical

PEG-10000

60 % v/v

100

Direct

Continuous

glycine 100 mM

8.6

37°C

? mM 1-Iodohexane

1-Hexanol, Iodide ion

—

Classical aqueous control (in %)

—

Activity - Classical

Dimethylformamide (DMF)

60 % v/v

100

Direct

Continuous

glycine 100 mM

8.6

37°C

? mM 1-Iodohexane

1-Hexanol, Iodide ion

—

Classical aqueous control (in %)

—

Activity - Classical

Dimethyl Sulfoxide (DMSO)

60 % v/v

100

Direct

Continuous

glycine 100 mM

8.6

37°C

? mM 1-Iodohexane

1-Hexanol, Iodide ion

—

Classical aqueous control (in %)

—

Activity - Classical

Methanol

60 % v/v

100

Direct

Continuous

glycine 100 mM

8.6

37°C

? mM 1-Iodohexane

1-Hexanol, Iodide ion

—

Classical aqueous control (in %)

—

Activity - Classical

1,4-Dioxane

60 % v/v

100

Direct

Continuous

glycine 100 mM

8.6

37°C

? mM 1-Iodohexane

1-Hexanol, Iodide ion

—

Classical aqueous control (in %)

—

Activity - Classical

Acetonitrile

60 % v/v

100

Direct

Continuous

glycine 100 mM

8.6

37°C

? mM 1-Iodohexane

1-Hexanol, Iodide ion

—

Classical aqueous control (in %)

—

Activity - Classical

Ethanol

60 % v/v

100

Direct

Continuous

glycine 100 mM

8.6

37°C

? mM 1-Iodohexane

1-Hexanol, Iodide ion

—

Classical aqueous control (in %)

—

Activity - Classical

Acetone

60 % v/v

100

Direct

Continuous

glycine 100 mM

8.6

37°C

? mM 1-Iodohexane

1-Hexanol, Iodide ion

—

Classical aqueous control (in %)

—

Activity - Classical

Isopropanol

60 % v/v

100

Direct

Continuous

glycine 100 mM

8.6

37°C

? mM 1-Iodohexane

1-Hexanol, Iodide ion

—

Classical aqueous control (in %)

—

Activity - Classical

Tetrahydrofuran (THF)

60 % v/v

100

Direct

Continuous

glycine 100 mM

8.6

37°C

? mM 1-Iodohexane

1-Hexanol, Iodide ion

—

Classical aqueous control (in %)

—

Activity - Classical

Glycerol

75 % v/v

100

Direct

Continuous

glycine 100 mM

8.6

37°C

? mM 1-Iodohexane

1-Hexanol, Iodide ion

—

Classical aqueous control (in %)

—

Activity - Classical

Formamide

75 % v/v

100

Direct

Continuous

glycine 100 mM

8.6

37°C

? mM 1-Iodohexane

1-Hexanol, Iodide ion

—

Classical aqueous control (in %)

—

Activity - Classical

Ethylene Glycol

75 % v/v

100

Direct

Continuous

glycine 100 mM

8.6

37°C

? mM 1-Iodohexane

1-Hexanol, Iodide ion

—

Classical aqueous control (in %)

—

Activity - Classical

PEG-1000

75 % v/v

100

125

Direct

Continuous

glycine 100 mM

8.6

37°C

? mM 1-Iodohexane

1-Hexanol, Iodide ion

—

Classical aqueous control (in %)

—

Activity - Classical

PEG-10000

75 % v/v

100

Direct

Continuous

glycine 100 mM

8.6

37°C

? mM 1-Iodohexane

1-Hexanol, Iodide ion

—

Classical aqueous control (in %)

—

Activity - Classical

Dimethylformamide (DMF)

75 % v/v

100

Direct

Continuous

glycine 100 mM

8.6

37°C

? mM 1-Iodohexane

1-Hexanol, Iodide ion

—

Classical aqueous control (in %)

—

Activity - Classical

Dimethyl Sulfoxide (DMSO)

75 % v/v

100

Direct

Continuous

glycine 100 mM

8.6

37°C

? mM 1-Iodohexane

1-Hexanol, Iodide ion

—

Classical aqueous control (in %)

—

Activity - Classical

Methanol

75 % v/v

100

Direct

Continuous

glycine 100 mM

8.6

37°C

? mM 1-Iodohexane

1-Hexanol, Iodide ion

—

Classical aqueous control (in %)

—

Activity - Classical

1,4-Dioxane

75 % v/v

100

Direct

Continuous

glycine 100 mM

8.6

37°C

? mM 1-Iodohexane

1-Hexanol, Iodide ion

—

Classical aqueous control (in %)

—

Activity - Classical

Acetonitrile

75 % v/v

100

Direct

Continuous

glycine 100 mM

8.6

37°C

? mM 1-Iodohexane

1-Hexanol, Iodide ion

—

Classical aqueous control (in %)

—

Activity - Classical

Ethanol

75 % v/v

100

Direct

Continuous

glycine 100 mM

8.6

37°C

? mM 1-Iodohexane

1-Hexanol, Iodide ion

—

Classical aqueous control (in %)

—

Activity - Classical

Acetone

75 % v/v

100

Direct

Continuous

glycine 100 mM

8.6

37°C

? mM 1-Iodohexane

1-Hexanol, Iodide ion

—

Classical aqueous control (in %)

—

Activity - Classical

Isopropanol

75 % v/v

100

Direct

Continuous

glycine 100 mM

8.6

37°C

? mM 1-Iodohexane

1-Hexanol, Iodide ion

—

Classical aqueous control (in %)

—

Activity - Classical

Tetrahydrofuran (THF)

75 % v/v

100

Direct

Continuous

glycine 100 mM

8.6

37°C

? 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

—

% (v/v)

Direct

Continuous

glycine 100 mM

8.6

37°C

1-Iodohexane

1-Hexanol, Iodide ion

—

Not applicable control (in % (v/v))

—

Stability - C50

Formamide

—

% (v/v)

Direct

Continuous

glycine 100 mM

8.6

37°C

1-Iodohexane

1-Hexanol, Iodide ion

—

Not applicable control (in % (v/v))

—

Stability - C50

Ethylene Glycol

—

% (v/v)

Direct

Continuous

glycine 100 mM

8.6

37°C

1-Iodohexane

1-Hexanol, Iodide ion

—

Not applicable control (in % (v/v))

—

Stability - C50

Dimethyl Sulfoxide (DMSO)

—

% (v/v)

Direct

Continuous

glycine 100 mM

8.6

37°C

1-Iodohexane

1-Hexanol, Iodide ion

—

Not applicable control (in % (v/v))

—

Stability - C50

Dimethylformamide (DMF)

—

% (v/v)

Direct

Continuous

glycine 100 mM

8.6

37°C

1-Iodohexane

1-Hexanol, Iodide ion

—

Not applicable control (in % (v/v))

—

Stability - C50

Methanol

—

% (v/v)

Direct

Continuous

glycine 100 mM

8.6

37°C

1-Iodohexane

1-Hexanol, Iodide ion

—

Not applicable control (in % (v/v))

—

Stability - C50

Acetonitrile

—

% (v/v)

Direct

Continuous

glycine 100 mM

8.6

37°C

1-Iodohexane

1-Hexanol, Iodide ion

—

Not applicable control (in % (v/v))

—

Stability - C50

Ethanol

—

% (v/v)

Direct

Continuous

glycine 100 mM

8.6

37°C

1-Iodohexane

1-Hexanol, Iodide ion

—

Not applicable control (in % (v/v))

—

Stability - C50

1,4-Dioxane

—

% (v/v)

Direct

Continuous

glycine 100 mM

8.6

37°C

1-Iodohexane

1-Hexanol, Iodide ion

—

Not applicable control (in % (v/v))

—

Stability - C50

Acetone

—

% (v/v)

Direct

Continuous

glycine 100 mM

8.6

37°C

1-Iodohexane

1-Hexanol, Iodide ion

—

Not applicable control (in % (v/v))

—

Stability - C50

Isopropanol

—

% (v/v)

Direct

Continuous

glycine 100 mM

8.6

37°C

1-Iodohexane

1-Hexanol, Iodide ion

—

Not applicable control (in % (v/v))

—

Stability - C50

Tetrahydrofuran (THF)

—

% (v/v)

Direct

Continuous

glycine 100 mM

8.6

37°C

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 % v/v

°C

Direct

Continuous

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 % v/v

°C

Direct

Continuous

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 % v/v

°C

Direct

Continuous

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 % v/v

°C

Direct

Continuous

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 % v/v

°C

Direct

Continuous

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 % v/v

°C

Direct

Continuous

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 % v/v

<10

°C

Direct

Continuous

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 % v/v

°C

Direct

Continuous

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 % v/v

°C

Direct

Continuous

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 % v/v

°C

Direct

Continuous

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 % v/v

°C

Direct

Continuous

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 % v/v

°C

Direct

Continuous

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	Extraction method	Data type	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 % v/v	100.0	—	2.19	%	Digital	Continuous	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 % v/v	28.67	—	2.25	1/s	Direct	Continuous	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 % v/v	8.06	—	0.16	1/(s * mM)	Direct	Continuous	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 % v/v	3.56	—	14.17	mM	Direct	Continuous	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	%	Direct	Continuous	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	%	Direct	Continuous	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 % v/v	3.56	—	0.0	mM	Direct	Continuous	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 % v/v	51.9	—	39.0	°C	Direct	Continuous	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 % v/v	51.9	—	36.8	°C	Direct	Continuous	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 % v/v	51.9	—	30.6	°C	Direct	Continuous	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 % v/v	100.0	2.19	0.44	%	Digital	Continuous	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 % v/v	100.0	2.19	1.46	%	Digital	Continuous	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 % v/v	100.0	2.19	0.66	%	Digital	Continuous	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 % v/v	100.0	2.19	10.4	%	Digital	Continuous	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 % v/v	0.34	2.25	0.25	1/s	Direct	Continuous	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 % v/v	9.88	2.25	0.5	1/s	Direct	Continuous	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 % v/v	13.83	2.25	1.01	1/s	Direct	Continuous	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 % v/v	2.23	2.25	0.72	1/s	Direct	Continuous	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 % v/v	0.26	2.25	0.36	1/s	Direct	Continuous	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 % v/v	0.28	2.25	0.86	1/s	Direct	Continuous	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 % v/v	100.0	—	2.19	%	Digital	Continuous	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 % v/v	28.67	—	2.25	1/s	Direct	Continuous	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 % v/v	8.06	—	0.16	1/(s * mM)	Direct	Continuous	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 % v/v	3.56	—	14.17	mM	Direct	Continuous	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	%	Direct	Continuous	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	%	Direct	Continuous	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 % v/v	3.56	—	0.0	mM	Direct	Continuous	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 % v/v	51.9	—	39.0	°C	Direct	Continuous	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 % v/v	51.9	—	36.8	°C	Direct	Continuous	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 % v/v	51.9	—	30.6	°C	Direct	Continuous	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 % v/v	100.0	2.19	0.44	%	Digital	Continuous	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 % v/v	100.0	2.19	1.46	%	Digital	Continuous	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 % v/v	100.0	2.19	0.66	%	Digital	Continuous	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 % v/v	100.0	2.19	10.4	%	Digital	Continuous	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 % v/v	0.34	2.25	0.25	1/s	Direct	Continuous	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 % v/v	9.88	2.25	0.5	1/s	Direct	Continuous	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 % v/v	13.83	2.25	1.01	1/s	Direct	Continuous	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 % v/v	2.23	2.25	0.72	1/s	Direct	Continuous	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 % v/v	0.26	2.25	0.36	1/s	Direct	Continuous	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 % v/v	0.28	2.25	0.86	1/s	Direct	Continuous	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 % v/v	11.66	0.16	0.12	1/(s * mM)	Direct	Continuous	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 % v/v	11.79	0.16	0.18	1/(s * mM)	Direct	Continuous	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 % v/v	1.31	0.16	0.67	1/(s * mM)	Direct	Continuous	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 % v/v	2.49	0.16	0.29	1/(s * mM)	Direct	Continuous	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 % v/v	2.46	0.16	0.26	1/(s * mM)	Direct	Continuous	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 % v/v	2.45	0.16	0.17	1/(s * mM)	Direct	Continuous	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 % v/v	0.11	14.17	5.1	mM	Direct	Continuous	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 % v/v	0.11	14.17	1.4	mM	Direct	Continuous	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 % v/v	0.13	14.17	0.88	mM	Direct	Continuous	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 % v/v	1.7	14.17	1.08	mM	Direct	Continuous	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 % v/v	1.17	14.17	5.47	mM	Direct	Continuous	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 % v/v	0.85	14.17	4.09	mM	Direct	Continuous	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	%	Direct	Continuous	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	%	Direct	Continuous	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	%	Direct	Continuous	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	%	Direct	Continuous	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	%	Direct	Continuous	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	%	Direct	Continuous	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	%	Direct	Continuous	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	%	Direct	Continuous	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	%	Direct	Continuous	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	%	Direct	Continuous	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	%	Direct	Continuous	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	%	Direct	Continuous	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	%	Direct	Continuous	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	%	Direct	Continuous	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	%	Direct	Continuous	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	%	Direct	Continuous	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 % v/v	0.42	0.0	6.69	mM	Direct	Continuous	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 % v/v	0.41	0.0	13.14	mM	Direct	Continuous	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 % v/v	0.48	0.0	2.34	mM	Direct	Continuous	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 % v/v	0.24	0.0	41.44	mM	Direct	Continuous	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 % v/v	3.45	0.0	0.0	mM	Direct	Continuous	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 % v/v	6.55	0.0	0.0	mM	Direct	Continuous	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 % v/v	56.2	36.8	44.4	°C	Direct	Continuous	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 % v/v	68.2	30.6	42.2	°C	Direct	Continuous	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 % v/v	68.2	39.0	60.4	°C	Direct	Continuous	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 % v/v	68.2	36.8	56.5	°C	Direct	Continuous	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 % v/v	56.2	39.0	48.3	°C	Direct	Continuous	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 % v/v	65.9	30.6	42.1	°C	Direct	Continuous	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 % v/v	65.9	39.0	57.3	°C	Direct	Continuous	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 % v/v	65.9	36.8	52.0	°C	Direct	Continuous	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 % v/v	63.8	30.6	41.8	°C	Direct	Continuous	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 % v/v	63.8	39.0	54.8	°C	Direct	Continuous	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 % v/v	63.8	36.8	50.5	°C	Direct	Continuous	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 % v/v	56.2	30.6	36.9	°C	Direct	Continuous	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

Visualization : Activity — Classical

Tana Koudelakova et al. (2013)

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

Tana Koudelakova et al. (2013)

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.

Visualization : Stability — T_m

Tana Koudelakova et al. (2013)

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 : Stability — Inactivation Rate Constant

Tana Koudelakova et al. (2013)

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 : Stability — C₅₀

Tana Koudelakova et al. (2013)

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

Structure

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Haloalkane dehalogenase DhaA from Rhodococcus rhodochrous

Enzyme Description

Sequence

Experimental Data (136 measurements)

Visualization : Activity — Classical

Visualization : Stability — Tm

Visualization : Stability — C50

Experimental Data (66 measurements)

Visualization : Activity — Classical

Mutation Effect

Visualization : Activity — Michaelis-Menten

Mutation Effect — Michaelis-Menten

Visualization : Stability — Tm

Mutation Effect

Visualization : Stability — Inactivation Rate Constant

Mutation Effect

Visualization : Stability — C50

Structure

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Visualization : Stability — T_m

Visualization : Stability — C₅₀

Visualization : Stability — T_m

Visualization : Stability — C₅₀