Phenylacetone monooxygenase from Thermobifida fusca

Enzyme Description

Species

Thermobifida fusca ↗

Extremophile

Yes thermophilic organism

EC Number

1.14.13.92 ↗, 1.14.13.92 ↗ (sulfoxidation)

Sequence

Length: 542 amino acids

MAGQTTVDSRRQPPEEVDVLVVGAGFSGLYALYRLRELGRSVHVIETAGDVGGVWYWNRYPGARCDIESIEYCYSFSEEVLQEWNWTERYASQPEILRYINFVADKFDLRSGITFHTTVTAAAFDEATNTWTVDTNHGDRIRARYLIMASGQLSVPQLPNFPGLKDFAGNLYHTGNWPHEPVDFSGQRVGVIGTGSSGIQVSPQIAKQAAELFVFQRTPHFAVPARNAPLDPEFLADLKKRYAEFREESRNTPGGTHRYQGPKSALEVSDEELVETLERYWQEGGPDILAAYRDILRDRDANERVAEFIRNKIRNTVRDPEVAERLVPKGYPFGTKRLILEIDYYEMFNRDNVHLVDTLSAPIETITPRGVRTSEREYELDSLVLATGFDALTGALFKIDIRGVGNVALKEKWAAGPRTYLGLSTAGFPNLFFIAGPGSPSALSNMLVSIEQHVEWVTDHIAYMFKNGLTRSEAVLEKEDEWVEHVNEIADETLYPMTASWYTGANVPGKPRVFMLYVGGFHRYRQICDEVAAKGYEGFVLT

Francesco Secundo et al. (2010) — Effects of water miscible organic solvents on the activity and conformation of the baeyer–villiger monooxygenases from Thermobifida fusca and Acinetobacter calcoaceticus : A comparative study

Biotechnology and Bioengineering · doi:10.1002/bit.22963 ↗ · Activity - Classical Stability - C50 Activity + Stability - Incubation

39 measurements ▼

Database ID

UniProt: Q47PU3 ↗

Sequence Annotation

Inferred - from protein name

Protein Source

Recombinant, host bacterium Escherichia coli BL21

Experimental Data (39 measurements)

39 measurements — page 1 of 2

Property	Assay	Solvent	Solvent Volume	Aqueous Reference	Measured Value	Units	Solution	pH	Temperature	Substrate(s)	Product(s)	Cofactor(s)	Shaking	Comments
Activity - Classical	Activity (in U/mg) measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in the presence of organic solvent	Acetonitrile	20%	0.1	0.054	U/mg	50 mM Tris-HCl	8.6	25°C	Molecular oxygen (O2), Thioanisole	H2O , methyl phenyl sulfoxide	0.1 mM NADPH	—	Classical aqueous control (in U/mg)
Activity - Classical	Activity (in U/mg) measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in the presence of organic solvent	2,2,2-Trifluoroethanol (TFE)	60%	0.1	0.0	U/mg	50 mM Tris-HCl	8.6	25°C	Molecular oxygen (O2), Thioanisole	H2O , methyl phenyl sulfoxide	0.1 mM NADPH	—	Classical aqueous control (in U/mg)
Activity - Classical	Activity (in U/mg) measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in the presence of organic solvent	2,2,2-Trifluoroethanol (TFE)	50%	0.1	0.0	U/mg	50 mM Tris-HCl	8.6	25°C	Molecular oxygen (O2), Thioanisole	H2O , methyl phenyl sulfoxide	0.1 mM NADPH	—	Classical aqueous control (in U/mg)
Activity - Classical	Activity (in U/mg) measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in the presence of organic solvent	2,2,2-Trifluoroethanol (TFE)	40%	0.1	0.0	U/mg	50 mM Tris-HCl	8.6	25°C	Molecular oxygen (O2), Thioanisole	H2O , methyl phenyl sulfoxide	0.1 mM NADPH	—	Classical aqueous control (in U/mg)
Activity - Classical	Activity (in U/mg) measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in the presence of organic solvent	2,2,2-Trifluoroethanol (TFE)	30%	0.1	0.0	U/mg	50 mM Tris-HCl	8.6	25°C	Molecular oxygen (O2), Thioanisole	H2O , methyl phenyl sulfoxide	0.1 mM NADPH	—	Classical aqueous control (in U/mg)
Activity - Classical	Activity (in U/mg) measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in the presence of organic solvent	2,2,2-Trifluoroethanol (TFE)	20%	0.1	0.0	U/mg	50 mM Tris-HCl	8.6	25°C	Molecular oxygen (O2), Thioanisole	H2O , methyl phenyl sulfoxide	0.1 mM NADPH	—	Classical aqueous control (in U/mg)
Activity - Classical	Activity (in U/mg) measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in the presence of organic solvent	2,2,2-Trifluoroethanol (TFE)	15%	0.1	0.06	U/mg	50 mM Tris-HCl	8.6	25°C	Molecular oxygen (O2), Thioanisole	H2O , methyl phenyl sulfoxide	0.1 mM NADPH	—	Classical aqueous control (in U/mg)
Activity - Classical	Activity (in U/mg) measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in the presence of organic solvent	2,2,2-Trifluoroethanol (TFE)	10%	0.1	0.011	U/mg	50 mM Tris-HCl	8.6	25°C	Molecular oxygen (O2), Thioanisole	H2O , methyl phenyl sulfoxide	0.1 mM NADPH	—	Classical aqueous control (in U/mg)
Activity - Classical	Activity (in U/mg) measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in the presence of organic solvent	1,4-Dioxane	60%	0.1	0.004	U/mg	50 mM Tris-HCl	8.6	25°C	Molecular oxygen (O2), Thioanisole	H2O , methyl phenyl sulfoxide	0.1 mM NADPH	—	Classical aqueous control (in U/mg)
Activity - Classical	Activity (in U/mg) measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in the presence of organic solvent	1,4-Dioxane	50%	0.1	0.005	U/mg	50 mM Tris-HCl	8.6	25°C	Molecular oxygen (O2), Thioanisole	H2O , methyl phenyl sulfoxide	0.1 mM NADPH	—	Classical aqueous control (in U/mg)
Activity - Classical	Activity (in U/mg) measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in the presence of organic solvent	1,4-Dioxane	35%	0.1	0.02	U/mg	50 mM Tris-HCl	8.6	25°C	Molecular oxygen (O2), Thioanisole	H2O , methyl phenyl sulfoxide	0.1 mM NADPH	—	Classical aqueous control (in U/mg)
Activity - Classical	Activity (in U/mg) measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in the presence of organic solvent	1,4-Dioxane	25%	0.1	0.05	U/mg	50 mM Tris-HCl	8.6	25°C	Molecular oxygen (O2), Thioanisole	H2O , methyl phenyl sulfoxide	0.1 mM NADPH	—	Classical aqueous control (in U/mg)
Activity - Classical	Activity (in U/mg) measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in the presence of organic solvent	1,4-Dioxane	15%	0.1	0.09	U/mg	50 mM Tris-HCl	8.6	25°C	Molecular oxygen (O2), Thioanisole	H2O , methyl phenyl sulfoxide	0.1 mM NADPH	—	Classical aqueous control (in U/mg)
Activity - Classical	Activity (in U/mg) measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in the presence of organic solvent	1,4-Dioxane	10%	0.1	0.09	U/mg	50 mM Tris-HCl	8.6	25°C	Molecular oxygen (O2), Thioanisole	H2O , methyl phenyl sulfoxide	0.1 mM NADPH	—	Classical aqueous control (in U/mg)
Activity - Classical	Activity (in U/mg) measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in the presence of organic solvent	1,4-Dioxane	5%	0.1	0.08	U/mg	50 mM Tris-HCl	8.6	25°C	Molecular oxygen (O2), Thioanisole	H2O , methyl phenyl sulfoxide	0.1 mM NADPH	—	Classical aqueous control (in U/mg)
Activity - Classical	Activity (in U/mg) measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in the presence of organic solvent	Ethanol	60%	0.1	0.01	U/mg	50 mM Tris-HCl	8.6	25°C	Molecular oxygen (O2), Thioanisole	H2O , methyl phenyl sulfoxide	0.1 mM NADPH	—	Classical aqueous control (in U/mg)
Activity - Classical	Activity (in U/mg) measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in the presence of organic solvent	Ethanol	40%	0.1	0.06	U/mg	50 mM Tris-HCl	8.6	25°C	Molecular oxygen (O2), Thioanisole	H2O , methyl phenyl sulfoxide	0.1 mM NADPH	—	Classical aqueous control (in U/mg)
Activity - Classical	Activity (in U/mg) measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in the presence of organic solvent	Acetonitrile	10%	0.1	0.1	U/mg	50 mM Tris-HCl	8.6	25°C	Molecular oxygen (O2), Thioanisole	H2O , methyl phenyl sulfoxide	0.1 mM NADPH	—	Classical aqueous control (in U/mg)
Activity - Classical	Activity (in U/mg) measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in the presence of organic solvent	Acetonitrile	30%	0.1	0.015	U/mg	50 mM Tris-HCl	8.6	25°C	Molecular oxygen (O2), Thioanisole	H2O , methyl phenyl sulfoxide	0.1 mM NADPH	—	Classical aqueous control (in U/mg)
Activity - Classical	Activity (in U/mg) measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in the presence of organic solvent	Acetonitrile	40%	0.1	0.015	U/mg	50 mM Tris-HCl	8.6	25°C	Molecular oxygen (O2), Thioanisole	H2O , methyl phenyl sulfoxide	0.1 mM NADPH	—	Classical aqueous control (in U/mg)

—

Activity - Classical

Activity (in U/mg) measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in the presence of organic solvent

Acetonitrile

20%

0.1

0.054

U/mg

50 mM Tris-HCl

8.6

25°C

Molecular oxygen (O2), Thioanisole

H2O, methyl phenyl sulfoxide

0.1 mM NADPH

—

Classical aqueous control (in U/mg)

—

Activity - Classical

Activity (in U/mg) measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in the presence of organic solvent

2,2,2-Trifluoroethanol (TFE)

60%

0.1

0.0

U/mg

50 mM Tris-HCl

8.6

25°C

Molecular oxygen (O2), Thioanisole

H2O, methyl phenyl sulfoxide

0.1 mM NADPH

—

Classical aqueous control (in U/mg)

—

Activity - Classical

Activity (in U/mg) measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in the presence of organic solvent

2,2,2-Trifluoroethanol (TFE)

50%

0.1

0.0

U/mg

50 mM Tris-HCl

8.6

25°C

Molecular oxygen (O2), Thioanisole

H2O, methyl phenyl sulfoxide

0.1 mM NADPH

—

Classical aqueous control (in U/mg)

—

Activity - Classical

Activity (in U/mg) measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in the presence of organic solvent

2,2,2-Trifluoroethanol (TFE)

40%

0.1

0.0

U/mg

50 mM Tris-HCl

8.6

25°C

Molecular oxygen (O2), Thioanisole

H2O, methyl phenyl sulfoxide

0.1 mM NADPH

—

Classical aqueous control (in U/mg)

—

Activity - Classical

Activity (in U/mg) measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in the presence of organic solvent

2,2,2-Trifluoroethanol (TFE)

30%

0.1

0.0

U/mg

50 mM Tris-HCl

8.6

25°C

Molecular oxygen (O2), Thioanisole

H2O, methyl phenyl sulfoxide

0.1 mM NADPH

—

Classical aqueous control (in U/mg)

—

Activity - Classical

Activity (in U/mg) measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in the presence of organic solvent

2,2,2-Trifluoroethanol (TFE)

20%

0.1

0.0

U/mg

50 mM Tris-HCl

8.6

25°C

Molecular oxygen (O2), Thioanisole

H2O, methyl phenyl sulfoxide

0.1 mM NADPH

—

Classical aqueous control (in U/mg)

—

Activity - Classical

Activity (in U/mg) measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in the presence of organic solvent

2,2,2-Trifluoroethanol (TFE)

15%

0.1

0.06

U/mg

50 mM Tris-HCl

8.6

25°C

Molecular oxygen (O2), Thioanisole

H2O, methyl phenyl sulfoxide

0.1 mM NADPH

—

Classical aqueous control (in U/mg)

—

Activity - Classical

Activity (in U/mg) measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in the presence of organic solvent

2,2,2-Trifluoroethanol (TFE)

10%

0.1

0.011

U/mg

50 mM Tris-HCl

8.6

25°C

Molecular oxygen (O2), Thioanisole

H2O, methyl phenyl sulfoxide

0.1 mM NADPH

—

Classical aqueous control (in U/mg)

—

Activity - Classical

Activity (in U/mg) measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in the presence of organic solvent

1,4-Dioxane

60%

0.1

0.004

U/mg

50 mM Tris-HCl

8.6

25°C

Molecular oxygen (O2), Thioanisole

H2O, methyl phenyl sulfoxide

0.1 mM NADPH

—

Classical aqueous control (in U/mg)

—

Activity - Classical

Activity (in U/mg) measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in the presence of organic solvent

1,4-Dioxane

50%

0.1

0.005

U/mg

50 mM Tris-HCl

8.6

25°C

Molecular oxygen (O2), Thioanisole

H2O, methyl phenyl sulfoxide

0.1 mM NADPH

—

Classical aqueous control (in U/mg)

—

Activity - Classical

Activity (in U/mg) measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in the presence of organic solvent

1,4-Dioxane

35%

0.1

0.02

U/mg

50 mM Tris-HCl

8.6

25°C

Molecular oxygen (O2), Thioanisole

H2O, methyl phenyl sulfoxide

0.1 mM NADPH

—

Classical aqueous control (in U/mg)

—

Activity - Classical

Activity (in U/mg) measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in the presence of organic solvent

1,4-Dioxane

25%

0.1

0.05

U/mg

50 mM Tris-HCl

8.6

25°C

Molecular oxygen (O2), Thioanisole

H2O, methyl phenyl sulfoxide

0.1 mM NADPH

—

Classical aqueous control (in U/mg)

—

Activity - Classical

Activity (in U/mg) measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in the presence of organic solvent

1,4-Dioxane

15%

0.1

0.09

U/mg

50 mM Tris-HCl

8.6

25°C

Molecular oxygen (O2), Thioanisole

H2O, methyl phenyl sulfoxide

0.1 mM NADPH

—

Classical aqueous control (in U/mg)

—

Activity - Classical

Activity (in U/mg) measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in the presence of organic solvent

1,4-Dioxane

10%

0.1

0.09

U/mg

50 mM Tris-HCl

8.6

25°C

Molecular oxygen (O2), Thioanisole

H2O, methyl phenyl sulfoxide

0.1 mM NADPH

—

Classical aqueous control (in U/mg)

—

Activity - Classical

Activity (in U/mg) measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in the presence of organic solvent

1,4-Dioxane

0.1

0.08

U/mg

50 mM Tris-HCl

8.6

25°C

Molecular oxygen (O2), Thioanisole

H2O, methyl phenyl sulfoxide

0.1 mM NADPH

—

Classical aqueous control (in U/mg)

—

Activity - Classical

Activity (in U/mg) measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in the presence of organic solvent

Ethanol

60%

0.1

0.01

U/mg

50 mM Tris-HCl

8.6

25°C

Molecular oxygen (O2), Thioanisole

H2O, methyl phenyl sulfoxide

0.1 mM NADPH

—

Classical aqueous control (in U/mg)

—

Activity - Classical

Activity (in U/mg) measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in the presence of organic solvent

Ethanol

40%

0.1

0.06

U/mg

50 mM Tris-HCl

8.6

25°C

Molecular oxygen (O2), Thioanisole

H2O, methyl phenyl sulfoxide

0.1 mM NADPH

—

Classical aqueous control (in U/mg)

—

Activity - Classical

Activity (in U/mg) measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in the presence of organic solvent

Acetonitrile

10%

0.1

U/mg

50 mM Tris-HCl

8.6

25°C

Molecular oxygen (O2), Thioanisole

H2O, methyl phenyl sulfoxide

0.1 mM NADPH

—

Classical aqueous control (in U/mg)

—

Activity - Classical

Activity (in U/mg) measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in the presence of organic solvent

Acetonitrile

30%

0.1

0.015

U/mg

50 mM Tris-HCl

8.6

25°C

Molecular oxygen (O2), Thioanisole

H2O, methyl phenyl sulfoxide

0.1 mM NADPH

—

Classical aqueous control (in U/mg)

—

Activity - Classical

Activity (in U/mg) measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in the presence of organic solvent

Acetonitrile

40%

0.1

0.015

U/mg

50 mM Tris-HCl

8.6

25°C

Molecular oxygen (O2), Thioanisole

H2O, methyl phenyl sulfoxide

0.1 mM NADPH

—

Classical aqueous control (in U/mg)

—

Activity - Classical

Activity (in U/mg) measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in the presence of organic solvent

Acetonitrile

50%

0.1

0.005

U/mg

50 mM Tris-HCl

8.6

25°C

Molecular oxygen (O2), Thioanisole

H2O, methyl phenyl sulfoxide

0.1 mM NADPH

—

Classical aqueous control (in U/mg)

—

Activity - Classical

Activity (in U/mg) measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in the presence of organic solvent

Acetonitrile

60%

0.1

0.004

U/mg

50 mM Tris-HCl

8.6

25°C

Molecular oxygen (O2), Thioanisole

H2O, methyl phenyl sulfoxide

0.1 mM NADPH

—

Classical aqueous control (in U/mg)

—

Activity - Classical

Activity (in U/mg) measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in the presence of organic solvent

Methanol

10%

0.1

0.28

U/mg

50 mM Tris-HCl

8.6

25°C

Molecular oxygen (O2), Thioanisole

H2O, methyl phenyl sulfoxide

0.1 mM NADPH

—

Classical aqueous control (in U/mg)

—

Activity - Classical

Activity (in U/mg) measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in the presence of organic solvent

Methanol

20%

0.1

0.46

U/mg

50 mM Tris-HCl

8.6

25°C

Molecular oxygen (O2), Thioanisole

H2O, methyl phenyl sulfoxide

0.1 mM NADPH

—

Classical aqueous control (in U/mg)

—

Activity - Classical

Activity (in U/mg) measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in the presence of organic solvent

Methanol

30%

0.1

0.25

U/mg

50 mM Tris-HCl

8.6

25°C

Molecular oxygen (O2), Thioanisole

H2O, methyl phenyl sulfoxide

0.1 mM NADPH

—

Classical aqueous control (in U/mg)

—

Activity - Classical

Activity (in U/mg) measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in the presence of organic solvent

Methanol

40%

0.1

0.15

U/mg

50 mM Tris-HCl

8.6

25°C

Molecular oxygen (O2), Thioanisole

H2O, methyl phenyl sulfoxide

0.1 mM NADPH

—

Classical aqueous control (in U/mg)

—

Activity - Classical

Activity (in U/mg) measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in the presence of organic solvent

Methanol

50%

0.1

0.06

U/mg

50 mM Tris-HCl

8.6

25°C

Molecular oxygen (O2), Thioanisole

H2O, methyl phenyl sulfoxide

0.1 mM NADPH

—

Classical aqueous control (in U/mg)

—

Activity - Classical

Activity (in U/mg) measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in the presence of organic solvent

Methanol

60%

0.1

0.02

U/mg

50 mM Tris-HCl

8.6

25°C

Molecular oxygen (O2), Thioanisole

H2O, methyl phenyl sulfoxide

0.1 mM NADPH

—

Classical aqueous control (in U/mg)

—

Activity - Classical

Activity (in U/mg) measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in the presence of organic solvent

Ethanol

10%

0.1

0.011

U/mg

50 mM Tris-HCl

8.6

25°C

Molecular oxygen (O2), Thioanisole

H2O, methyl phenyl sulfoxide

0.1 mM NADPH

—

Classical aqueous control (in U/mg)

—

Activity - Classical

Activity (in U/mg) measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in the presence of organic solvent

Ethanol

20%

0.1

0.01

U/mg

50 mM Tris-HCl

8.6

25°C

Molecular oxygen (O2), Thioanisole

H2O, methyl phenyl sulfoxide

0.1 mM NADPH

—

Classical aqueous control (in U/mg)

—

Activity - Classical

Activity (in U/mg) measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in the presence of organic solvent

Ethanol

30%

0.1

0.09

U/mg

50 mM Tris-HCl

8.6

25°C

Molecular oxygen (O2), Thioanisole

H2O, methyl phenyl sulfoxide

0.1 mM NADPH

—

Classical aqueous control (in U/mg)

—

Activity - Classical

Activity (in U/mg) measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in the presence of organic solvent

Ethanol

50%

0.1

0.02

U/mg

50 mM Tris-HCl

8.6

25°C

Molecular oxygen (O2), Thioanisole

H2O, methyl phenyl sulfoxide

0.1 mM NADPH

—

Classical aqueous control (in U/mg)

—

Activity + Stability - Incubation

Activity after incubation (24 hours at 25°C) measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in the presence of organic solvent

Methanol

40%

0.088

0.004

U/mg

Incubation: 50 mM Tris-HCl, Assay: 50 mM Tris-HCl

Incubation: 8.6, Assay: 8.6

Incubation: 25°C, Assay: 25°C

Molecular oxygen (O2), Thioanisole

H2O, methyl phenyl sulfoxide

0.1 mM NADPH

—

Water-incubated control in water (in U/mg), assay in organic solvent

—

Activity + Stability - Incubation

Activity after incubation (24 hours at 25°C) measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in the presence of organic solvent

Methanol

20%

0.088

0.43

U/mg

Incubation: 50 mM Tris-HCl, Assay: 50 mM Tris-HCl

Incubation: 8.6, Assay: 8.6

Incubation: 25°C, Assay: 25°C

Molecular oxygen (O2), Thioanisole

H2O, methyl phenyl sulfoxide

0.1 mM NADPH

—

Water-incubated control in water (in U/mg), assay in organic solvent

—

Stability - C50

Concentration (in % v/v) for which 50% of enzyme activity is lost, as measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm)

Methanol

—

55.0

% (v/v)

50 mM Tris-HCl

8.6

25°C

Molecular oxygen (O2), Thioanisole

H2O, methyl phenyl sulfoxide

0.1 mM NADPH

—

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

—

Stability - C50

Concentration (in % v/v) for which 50% of enzyme activity is lost, as measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm)

Ethanol

—

43.0

% (v/v)

50 mM Tris-HCl

8.6

25°C

Molecular oxygen (O2), Thioanisole

H2O, methyl phenyl sulfoxide

0.1 mM NADPH

—

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

—

Stability - C50

Concentration (in % v/v) for which 50% of enzyme activity is lost, as measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm)

1,4-Dioxane

—

26.0

% (v/v)

50 mM Tris-HCl

8.6

25°C

Molecular oxygen (O2), Thioanisole

H2O, methyl phenyl sulfoxide

0.1 mM NADPH

—

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

—

Stability - C50

Concentration (in % v/v) for which 50% of enzyme activity is lost, as measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm)

Acetonitrile

—

22.0

% (v/v)

50 mM Tris-HCl

8.6

25°C

Molecular oxygen (O2), Thioanisole

H2O, methyl phenyl sulfoxide

0.1 mM NADPH

—

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

—

Stability - C50

Concentration (in % v/v) for which 50% of enzyme activity is lost, as measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm)

2,2,2-Trifluoroethanol (TFE)

—

16.0

% (v/v)

50 mM Tris-HCl

8.6

25°C

Molecular oxygen (O2), Thioanisole

H2O, methyl phenyl sulfoxide

0.1 mM NADPH

—

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

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Visualization : Activity — Classical

Francesco Secundo et al. (2010)

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

Visualization : Activity + Stability — Incubation

Francesco Secundo et al. (2010)

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

Visualization : Stability — C₅₀

Francesco Secundo et al. (2010)

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

Gonzalo de Gonzalo et al. (2012) — Improvement of the biocatalytic properties of one phenylacetone monooxygenase mutant in hydrophilic organic solvents

Enzyme and Microbial Technology · doi:10.1016/j.enzmictec.2011.09.006 ↗ · Activity - Classical Stability - Incubation Activity + Stability - Conversion Specificity - Enantioselectivity

67 measurements ▼

Database ID

UniProt: Q47PU3 ↗

Sequence Annotation

Inferred - from protein name (1 mutation)

Protein Source

Recombinant, host bacterium Escherichia coli BL21

Experimental Data (67 measurements)

67 measurements — page 4 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
M446G	Specificity - Enantioselectivity	Enantiomeric excess (toward the S form) after incubation (24 hours at 30°C) in the presence of organic solvent, measured by HPLC	Methanol	5%	79.8	—	84.8	%	50 mM Tris-HCl, 20 mM G6P, 5U G6PDH	9	30°C	10 mM methyl phenyl sulfoxide, O₂	(S)-para-nitrophenyl methyl sulfoxide	0.2 mM NADPH	250 rpm	Classical aqueous control (in %)
M446G	Stability - Incubation	Activity after incubation (24 hours at 25°C), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in the presence of organic solvent	PEG	5%	100.0	—	89.0	%	Incubation: 50 mM Tris-HCl, Assay: 50 mM Tris-HCl	9	30°C	Benzyl methyl sulfide, O₂	(S)-Benzyl dimethyl sulfoxonium	0.1 mM NADPH	Incubation: 250 rpm	Water-incubated control (in %), 100% value and assay in the presence of organic solvent
M446G	Stability - Incubation	Activity after incubation (24 hours at 25°C), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in the presence of organic solvent	Methanol	5%	100.0	—	84.0	%	Incubation: 50 mM Tris-HCl, Assay: 50 mM Tris-HCl	9	30°C	Benzyl methyl sulfide, O₂	(S)-Benzyl dimethyl sulfoxonium	0.1 mM NADPH	Incubation: 250 rpm	Water-incubated control (in %), 100% value and assay in the presence of organic solvent
M446G	Stability - Incubation	Activity after incubation (24 hours at 25°C), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in the presence of organic solvent	Isopropanol	5%	100.0	—	41.0	%	Incubation: 50 mM Tris-HCl, Assay: 50 mM Tris-HCl	9	30°C	Benzyl methyl sulfide, O₂	(S)-Benzyl dimethyl sulfoxonium	0.1 mM NADPH	Incubation: 250 rpm	Water-incubated control (in %), 100% value and assay in the presence of organic solvent
M446G	Stability - Incubation	Activity after incubation (24 hours at 25°C), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in the presence of organic solvent	PEG	30%	100.0	—	11.0	%	Incubation: 50 mM Tris-HCl, Assay: 50 mM Tris-HCl	9	30°C	Benzyl methyl sulfide, O₂	(S)-Benzyl dimethyl sulfoxonium	0.1 mM NADPH	Incubation: 250 rpm	Water-incubated control (in %), 100% value and assay in the presence of organic solvent
M446G	Stability - Incubation	Activity after incubation (24 hours at 25°C), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in the presence of organic solvent	Methanol	30%	100.0	—	7.0	%	Incubation: 50 mM Tris-HCl, Assay: 50 mM Tris-HCl	9	30°C	Benzyl methyl sulfide, O₂	(S)-Benzyl dimethyl sulfoxonium	0.1 mM NADPH	Incubation: 250 rpm	Water-incubated control (in %), 100% value and assay in the presence of organic solvent
M446G	Stability - Incubation	Activity after incubation (24 hours at 25°C), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in the presence of organic solvent	Isopropanol	30%	100.0	—	0.0	%	Incubation: 50 mM Tris-HCl, Assay: 50 mM Tris-HCl	9	30°C	Benzyl methyl sulfide, O₂	(S)-Benzyl dimethyl sulfoxonium	0.1 mM NADPH	Incubation: 250 rpm	Water-incubated control (in %), 100% value and assay in the presence of organic solvent

M446G

Activity - Classical

Activity (k_obs in s^-1) measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in the presence of organic solvent

Methanol

30%

2.9

—

1.5

1/s

50 mM Tris-HCl

30°C

Benzyl methyl sulfide, O₂

(S)-Benzyl dimethyl sulfoxonium

0.2 mM NADPH

—

Classical aqueous control (in 1/s)

M446G

Activity - Classical

Activity (k_obs in s^-1) measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in the presence of organic solvent

Isopropanol

10%

2.9

—

2.2

1/s

50 mM Tris-HCl

30°C

Benzyl methyl sulfide, O₂

(S)-Benzyl dimethyl sulfoxonium

0.2 mM NADPH

—

Classical aqueous control (in 1/s)

M446G

Activity - Classical

Activity (k_obs in s^-1) measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in the presence of organic solvent

Isopropanol

30%

2.9

—

0.24

1/s

50 mM Tris-HCl

30°C

Benzyl methyl sulfide, O₂

(S)-Benzyl dimethyl sulfoxonium

0.2 mM NADPH

—

Classical aqueous control (in 1/s)

M446G

Activity - Classical

Activity (k_obs in s^-1) measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in the presence of organic solvent

Acetonitrile

2.9

—

2.7

1/s

50 mM Tris-HCl

30°C

Benzyl methyl sulfide, O₂

(S)-Benzyl dimethyl sulfoxonium

0.2 mM NADPH

—

Classical aqueous control (in 1/s)

M446G

Activity - Classical

Activity (k_obs in s^-1) measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in the presence of organic solvent

Acetonitrile

10%

2.9

—

0.66

1/s

50 mM Tris-HCl

30°C

Benzyl methyl sulfide, O₂

(S)-Benzyl dimethyl sulfoxonium

0.2 mM NADPH

—

Classical aqueous control (in 1/s)

M446G

Activity - Classical

Activity (k_obs in s^-1) measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in the presence of organic solvent

Acetonitrile

30%

2.9

—

0.17

1/s

50 mM Tris-HCl

30°C

Benzyl methyl sulfide, O₂

(S)-Benzyl dimethyl sulfoxonium

0.2 mM NADPH

—

Classical aqueous control (in 1/s)

M446G

Activity - Classical

Activity (k_obs in s^-1) measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in the presence of organic solvent

Isopropanol

2.9

—

2.5

1/s

50 mM Tris-HCl

30°C

Benzyl methyl sulfide, O₂

(S)-Benzyl dimethyl sulfoxonium

0.2 mM NADPH

—

Classical aqueous control (in 1/s)

M446G

Activity - Classical

Activity (k_obs in s^-1) measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in the presence of organic solvent

PEG

50%

2.9

—

0.21

1/s

50 mM Tris-HCl

30°C

Benzyl methyl sulfide, O₂

(S)-Benzyl dimethyl sulfoxonium

0.2 mM NADPH

—

Classical aqueous control (in 1/s)

M446G

Activity - Classical

Activity (k_obs in s^-1) measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in the presence of organic solvent

PEG

30%

2.9

—

2.1

1/s

50 mM Tris-HCl

30°C

Benzyl methyl sulfide, O₂

(S)-Benzyl dimethyl sulfoxonium

0.2 mM NADPH

—

Classical aqueous control (in 1/s)

M446G

Activity - Classical

Activity (k_obs in s^-1) measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in the presence of organic solvent

PEG

10%

2.9

—

3.0

1/s

50 mM Tris-HCl

30°C

Benzyl methyl sulfide, O₂

(S)-Benzyl dimethyl sulfoxonium

0.2 mM NADPH

—

Classical aqueous control (in 1/s)

M446G

Activity - Classical

Activity (k_obs in s^-1) measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in the presence of organic solvent

PEG

2.9

—

3.2

1/s

50 mM Tris-HCl

30°C

Benzyl methyl sulfide, O₂

(S)-Benzyl dimethyl sulfoxonium

0.2 mM NADPH

—

Classical aqueous control (in 1/s)

M446G

Activity - Classical

Activity (k_obs in s^-1) measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in the presence of organic solvent

Methanol

10%

2.9

—

2.9

1/s

50 mM Tris-HCl

30°C

Benzyl methyl sulfide, O₂

(S)-Benzyl dimethyl sulfoxonium

0.2 mM NADPH

—

Classical aqueous control (in 1/s)

M446G

Activity - Classical

Activity (k_obs in s^-1) measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in the presence of organic solvent

Methanol

2.9

—

3.25

1/s

50 mM Tris-HCl

30°C

Benzyl methyl sulfide, O₂

(S)-Benzyl dimethyl sulfoxonium

0.2 mM NADPH

—

Classical aqueous control (in 1/s)

M446G

Activity + Stability - Conversion

Conversion after incubation (24 hours at 30°C) in the presence of organic solvent, measured by gas chromatography

Methanol

10%

81.1

—

97.9

50 mM Tris-HCl, 20 mM G6P, 5U G6PDH

30°C

Benzyl methyl sulfide, O₂

(S)-Benzyl dimethyl sulfoxonium

0.2 mM NADPH

250 rpm

Classical aqueous control (in %)

M446G

Activity + Stability - Conversion

Conversion after incubation (24 hours at 30°C) in the presence of organic solvent, measured by gas chromatography

PEG

10%

81.1

—

94.7

50 mM Tris-HCl, 20 mM G6P, 5U G6PDH

30°C

Benzyl methyl sulfide, O₂

(S)-Benzyl dimethyl sulfoxonium

0.2 mM NADPH

250 rpm

Classical aqueous control (in %)

M446G

Activity + Stability - Conversion

Conversion after incubation (24 hours at 30°C) in the presence of organic solvent, measured by gas chromatography

Methanol

78.1

—

96.2

50 mM Tris-HCl, 20 mM G6P, 5U G6PDH

30°C

O₂, 10 mM Thioanisole

(S)-methyl phenyl sulfoxide

0.2 mM NADPH

250 rpm

Classical aqueous control (in %)

M446G

Activity + Stability - Conversion

Conversion after incubation (24 hours at 30°C) in the presence of organic solvent, measured by gas chromatography

Methanol

34.9

—

70.2

50 mM Tris-HCl, 20 mM G6P, 5U G6PDH

30°C

O₂, 10 mM para-nitrophenyl methyl sulfide

(S)-para-nitrophenyl methyl sulfoxide

0.2 mM NADPH

250 rpm

Classical aqueous control (in %)

M446G

Activity + Stability - Conversion

Conversion after incubation (24 hours at 30°C) in the presence of organic solvent, measured by gas chromatography

Methanol

90.9

—

98.4

50 mM Tris-HCl, 20 mM G6P, 5U G6PDH

30°C

O₂, 10 mM Phenyl methyl sulfide

(S)-Phenyl methyl sulfoxide

0.2 mM NADPH

250 rpm

Classical aqueous control (in %)

M446G

Activity + Stability - Conversion

Conversion after incubation (24 hours at 30°C) in the presence of organic solvent, measured by gas chromatography

Methanol

62.4

—

96.4

50 mM Tris-HCl, 20 mM G6P, 5U G6PDH

30°C

10 mM Furfuryl methyl sulfide, O₂

(S)-Furfuryl methyl sulfoxide

0.2 mM NADPH

250 rpm

Classical aqueous control (in %)

M446G

Activity + Stability - Conversion

Conversion after incubation (24 hours at 30°C) in the presence of organic solvent, measured by gas chromatography

Methanol

68.9

—

71.2

50 mM Tris-HCl, 20 mM G6P, 5U G6PDH

30°C

10 mM Cyclohexyl methyl sulfide, O₂

(S)-Cyclohexyl methyl sulfoxide

0.2 mM NADPH

250 rpm

Classical aqueous control (in %)

M446G

Activity + Stability - Conversion

Conversion after incubation (24 hours at 30°C) in the presence of organic solvent, measured by gas chromatography

Methanol

17.9

—

36.1

50 mM Tris-HCl, 20 mM G6P, 5U G6PDH

30°C

10 mM Cyclohexyl ethyl sulfide, O₂

(S)-Cyclohexyl ethyl sulfoxide

0.2 mM NADPH

250 rpm

Classical aqueous control (in %)

M446G

Activity + Stability - Conversion

Conversion after incubation (24 hours at 30°C) in the presence of organic solvent, measured by gas chromatography

Methanol

<3.0

—

11.1

50 mM Tris-HCl, 20 mM G6P, 5U G6PDH

30°C

10 mM Cyclohexyl propyl sulfide, O₂

(S)-Cyclohexyl propyl sulfoxide

0.2 mM NADPH

250 rpm

Classical aqueous control (in %)

M446G

Activity + Stability - Conversion

Conversion after incubation (24 hours at 30°C) in the presence of organic solvent, measured by gas chromatography

Ethanol

30%

81.1

—

85.8

50 mM Tris-HCl, 20 mM G6P, 5U G6PDH

30°C

Benzyl methyl sulfide, O₂

(S)-Benzyl dimethyl sulfoxonium

0.2 mM NADPH

250 rpm

Classical aqueous control (in %)

M446G

Activity + Stability - Conversion

Conversion after incubation (24 hours at 30°C) in the presence of organic solvent, measured by gas chromatography

Acetonitrile

10%

81.1

—

>99.0

50 mM Tris-HCl, 20 mM G6P, 5U G6PDH

30°C

Benzyl methyl sulfide, O₂

(S)-Benzyl dimethyl sulfoxonium

0.2 mM NADPH

250 rpm

Classical aqueous control (in %)

M446G

Activity + Stability - Conversion

Conversion after incubation (24 hours at 30°C) in the presence of organic solvent, measured by gas chromatography

Ethanol

10%

81.1

—

>99.0

50 mM Tris-HCl, 20 mM G6P, 5U G6PDH

30°C

Benzyl methyl sulfide, O₂

(S)-Benzyl dimethyl sulfoxonium

0.2 mM NADPH

250 rpm

Classical aqueous control (in %)

M446G

Activity + Stability - Conversion

Conversion after incubation (24 hours at 30°C) in the presence of organic solvent, measured by gas chromatography

Isopropanol

10%

81.1

—

>99.0

50 mM Tris-HCl, 20 mM G6P, 5U G6PDH

30°C

Benzyl methyl sulfide, O₂

(S)-Benzyl dimethyl sulfoxonium

0.2 mM NADPH

250 rpm

Classical aqueous control (in %)

M446G

Activity + Stability - Conversion

Conversion after incubation (24 hours at 30°C) in the presence of organic solvent, measured by gas chromatography

Ethyl Acetate

10%

81.1

—

67.4

50 mM Tris-HCl, 20 mM G6P, 5U G6PDH

30°C

Benzyl methyl sulfide, O₂

(S)-Benzyl dimethyl sulfoxonium

0.2 mM NADPH

250 rpm

Classical aqueous control (in %)

M446G

Activity + Stability - Conversion

Conversion after incubation (24 hours at 30°C) in the presence of organic solvent, measured by gas chromatography

Diisopropyl Ether

10%

81.1

—

62.0

50 mM Tris-HCl, 20 mM G6P, 5U G6PDH

30°C

Benzyl methyl sulfide, O₂

(S)-Benzyl dimethyl sulfoxonium

0.2 mM NADPH

250 rpm

Classical aqueous control (in %)

M446G

Activity + Stability - Conversion

Conversion after incubation (24 hours at 30°C) in the presence of organic solvent, measured by gas chromatography

Toluene

10%

81.1

—

46.7

50 mM Tris-HCl, 20 mM G6P, 5U G6PDH

30°C

Benzyl methyl sulfide, O₂

(S)-Benzyl dimethyl sulfoxonium

0.2 mM NADPH

250 rpm

Classical aqueous control (in %)

M446G

Activity + Stability - Conversion

Conversion after incubation (24 hours at 30°C) in the presence of organic solvent, measured by gas chromatography

n-Hexane

10%

81.1

—

21.1

50 mM Tris-HCl, 20 mM G6P, 5U G6PDH

30°C

Benzyl methyl sulfide, O₂

(S)-Benzyl dimethyl sulfoxonium

0.2 mM NADPH

250 rpm

Classical aqueous control (in %)

M446G

Activity + Stability - Conversion

Conversion after incubation (24 hours at 30°C) in the presence of organic solvent, measured by gas chromatography

Isopropanol

30%

81.1

—

10.1

50 mM Tris-HCl, 20 mM G6P, 5U G6PDH

30°C

Benzyl methyl sulfide, O₂

(S)-Benzyl dimethyl sulfoxonium

0.2 mM NADPH

250 rpm

Classical aqueous control (in %)

M446G

Activity + Stability - Conversion

Conversion after incubation (24 hours at 30°C) in the presence of organic solvent, measured by gas chromatography

Methanol

81.1

—

95.1

50 mM Tris-HCl, 20 mM G6P, 5U G6PDH

30°C

Benzyl methyl sulfide, O₂

(S)-Benzyl dimethyl sulfoxonium

0.2 mM NADPH

250 rpm

Classical aqueous control (in %)

M446G

Activity + Stability - Conversion

Conversion after incubation (24 hours at 30°C) in the presence of organic solvent, measured by gas chromatography

Methanol

30%

81.1

—

82.3

50 mM Tris-HCl, 20 mM G6P, 5U G6PDH

30°C

Benzyl methyl sulfide, O₂

(S)-Benzyl dimethyl sulfoxonium

0.2 mM NADPH

250 rpm

Classical aqueous control (in %)

M446G

Activity + Stability - Conversion

Conversion after incubation (24 hours at 30°C) in the presence of organic solvent, measured by gas chromatography

Methanol

50%

81.1

—

4.3

50 mM Tris-HCl, 20 mM G6P, 5U G6PDH

30°C

Benzyl methyl sulfide, O₂

(S)-Benzyl dimethyl sulfoxonium

0.2 mM NADPH

250 rpm

Classical aqueous control (in %)

M446G

Activity + Stability - Conversion

Conversion after incubation (24 hours at 30°C) in the presence of organic solvent, measured by gas chromatography

Methanol

70%

81.1

—

<3.0

50 mM Tris-HCl, 20 mM G6P, 5U G6PDH

30°C

Benzyl methyl sulfide, O₂

(S)-Benzyl dimethyl sulfoxonium

0.2 mM NADPH

250 rpm

Classical aqueous control (in %)

M446G

Activity + Stability - Conversion

Conversion after incubation (24 hours at 30°C) in the presence of organic solvent, measured by gas chromatography

PEG

30%

81.1

—

73.5

50 mM Tris-HCl, 20 mM G6P, 5U G6PDH

30°C

Benzyl methyl sulfide, O₂

(S)-Benzyl dimethyl sulfoxonium

0.2 mM NADPH

250 rpm

Classical aqueous control (in %)

M446G

Activity + Stability - Conversion

Conversion after incubation (24 hours at 30°C) in the presence of organic solvent, measured by gas chromatography

Acetonitrile

30%

81.1

—

8.7

50 mM Tris-HCl, 20 mM G6P, 5U G6PDH

30°C

Benzyl methyl sulfide, O₂

(S)-Benzyl dimethyl sulfoxonium

0.2 mM NADPH

250 rpm

Classical aqueous control (in %)

M446G

Specificity - Enantioselectivity

Enantiomeric excess (toward the S form) after incubation (24 hours at 30°C) in the presence of organic solvent, measured by HPLC

Methanol

25.3

—

27.8

50 mM Tris-HCl, 20 mM G6P, 5U G6PDH

30°C

10 mM Furfuryl methyl sulfide, O₂

(S)-Furfuryl methyl sulfoxide

0.2 mM NADPH

250 rpm

Classical aqueous control (in %)

M446G

Specificity - Enantioselectivity

Enantiomeric excess (toward the S form) after incubation (24 hours at 30°C) in the presence of organic solvent, measured by HPLC

Isopropanol

30%

59.5

—

49.5

50 mM Tris-HCl, 20 mM G6P, 5U G6PDH

30°C

Benzyl methyl sulfide, O₂

(S)-Benzyl dimethyl sulfoxonium

0.2 mM NADPH

250 rpm

Classical aqueous control (in %)

M446G

Specificity - Enantioselectivity

Enantiomeric excess (toward the S form) after incubation (24 hours at 30°C) in the presence of organic solvent, measured by HPLC

Methanol

73.3

—

73.5

50 mM Tris-HCl, 20 mM G6P, 5U G6PDH

30°C

O₂, 10 mM Phenyl methyl sulfide

(S)-Phenyl methyl sulfoxide

0.2 mM NADPH

250 rpm

Classical aqueous control (in %)

M446G

Specificity - Enantioselectivity

Enantiomeric excess (toward the S form) after incubation (24 hours at 30°C) in the presence of organic solvent, measured by HPLC

Methanol

50.3

—

51.2

50 mM Tris-HCl, 20 mM G6P, 5U G6PDH

30°C

10 mM Cyclohexyl methyl sulfide, O₂

(S)-Cyclohexyl methyl sulfoxide

0.2 mM NADPH

250 rpm

Classical aqueous control (in %)

M446G

Specificity - Enantioselectivity

Enantiomeric excess (toward the S form) after incubation (24 hours at 30°C) in the presence of organic solvent, measured by HPLC

Methanol

30.3

—

36.7

50 mM Tris-HCl, 20 mM G6P, 5U G6PDH

30°C

10 mM Cyclohexyl ethyl sulfide, O₂

(S)-Cyclohexyl ethyl sulfoxide

0.2 mM NADPH

250 rpm

Classical aqueous control (in %)

M446G

Specificity - Enantioselectivity

Enantiomeric excess (toward the S form) after incubation (24 hours at 30°C) in the presence of organic solvent, measured by HPLC

Methanol

0.0

—

20.8

50 mM Tris-HCl, 20 mM G6P, 5U G6PDH

30°C

10 mM Cyclohexyl propyl sulfide, O₂

(S)-Cyclohexyl propyl sulfoxide

0.2 mM NADPH

250 rpm

Classical aqueous control (in %)

M446G

Specificity - Enantioselectivity

Enantiomeric excess (toward the S form) after incubation (24 hours at 30°C) in the presence of organic solvent, measured by HPLC

Methanol

89.8

—

93.3

50 mM Tris-HCl, 20 mM G6P, 5U G6PDH

30°C

O₂, 10 mM Thioanisole

(S)-methyl phenyl sulfoxide

0.2 mM NADPH

250 rpm

Classical aqueous control (in %)

M446G

Specificity - Enantioselectivity

Enantiomeric excess (toward the S form) after incubation (24 hours at 30°C) in the presence of organic solvent, measured by HPLC

PEG

10%

59.5

—

63.3

50 mM Tris-HCl, 20 mM G6P, 5U G6PDH

30°C

Benzyl methyl sulfide, O₂

(S)-Benzyl dimethyl sulfoxonium

0.2 mM NADPH

250 rpm

Classical aqueous control (in %)

M446G

Specificity - Enantioselectivity

Enantiomeric excess (toward the S form) after incubation (24 hours at 30°C) in the presence of organic solvent, measured by HPLC

Methanol

10%

59.5

—

59.5

50 mM Tris-HCl, 20 mM G6P, 5U G6PDH

30°C

Benzyl methyl sulfide, O₂

(S)-Benzyl dimethyl sulfoxonium

0.2 mM NADPH

250 rpm

Classical aqueous control (in %)

M446G

Specificity - Enantioselectivity

Enantiomeric excess (toward the S form) after incubation (24 hours at 30°C) in the presence of organic solvent, measured by HPLC

Acetonitrile

10%

59.5

—

57.3

50 mM Tris-HCl, 20 mM G6P, 5U G6PDH

30°C

Benzyl methyl sulfide, O₂

(S)-Benzyl dimethyl sulfoxonium

0.2 mM NADPH

250 rpm

Classical aqueous control (in %)

M446G

Specificity - Enantioselectivity

Enantiomeric excess (toward the S form) after incubation (24 hours at 30°C) in the presence of organic solvent, measured by HPLC

Ethanol

10%

59.5

—

61.3

50 mM Tris-HCl, 20 mM G6P, 5U G6PDH

30°C

Benzyl methyl sulfide, O₂

(S)-Benzyl dimethyl sulfoxonium

0.2 mM NADPH

250 rpm

Classical aqueous control (in %)

M446G

Specificity - Enantioselectivity

Enantiomeric excess (toward the S form) after incubation (24 hours at 30°C) in the presence of organic solvent, measured by HPLC

Isopropanol

10%

59.5

—

64.8

50 mM Tris-HCl, 20 mM G6P, 5U G6PDH

30°C

Benzyl methyl sulfide, O₂

(S)-Benzyl dimethyl sulfoxonium

0.2 mM NADPH

250 rpm

Classical aqueous control (in %)

M446G

Specificity - Enantioselectivity

Enantiomeric excess (toward the S form) after incubation (24 hours at 30°C) in the presence of organic solvent, measured by HPLC

Ethyl Acetate

10%

59.5

—

58.8

50 mM Tris-HCl, 20 mM G6P, 5U G6PDH

30°C

Benzyl methyl sulfide, O₂

(S)-Benzyl dimethyl sulfoxonium

0.2 mM NADPH

250 rpm

Classical aqueous control (in %)

M446G

Specificity - Enantioselectivity

Enantiomeric excess (toward the S form) after incubation (24 hours at 30°C) in the presence of organic solvent, measured by HPLC

Diisopropyl Ether

10%

59.5

—

56.0

50 mM Tris-HCl, 20 mM G6P, 5U G6PDH

30°C

Benzyl methyl sulfide, O₂

(S)-Benzyl dimethyl sulfoxonium

0.2 mM NADPH

250 rpm

Classical aqueous control (in %)

M446G

Specificity - Enantioselectivity

Enantiomeric excess (toward the S form) after incubation (24 hours at 30°C) in the presence of organic solvent, measured by HPLC

Toluene

10%

59.5

—

59.0

50 mM Tris-HCl, 20 mM G6P, 5U G6PDH

30°C

Benzyl methyl sulfide, O₂

(S)-Benzyl dimethyl sulfoxonium

0.2 mM NADPH

250 rpm

Classical aqueous control (in %)

M446G

Specificity - Enantioselectivity

Enantiomeric excess (toward the S form) after incubation (24 hours at 30°C) in the presence of organic solvent, measured by HPLC

n-Hexane

10%

59.5

—

58.8

50 mM Tris-HCl, 20 mM G6P, 5U G6PDH

30°C

Benzyl methyl sulfide, O₂

(S)-Benzyl dimethyl sulfoxonium

0.2 mM NADPH

250 rpm

Classical aqueous control (in %)

M446G

Specificity - Enantioselectivity

Enantiomeric excess (toward the S form) after incubation (24 hours at 30°C) in the presence of organic solvent, measured by HPLC

Methanol

59.5

—

62.5

50 mM Tris-HCl, 20 mM G6P, 5U G6PDH

30°C

Benzyl methyl sulfide, O₂

(S)-Benzyl dimethyl sulfoxonium

0.2 mM NADPH

250 rpm

Classical aqueous control (in %)

M446G

Specificity - Enantioselectivity

Enantiomeric excess (toward the S form) after incubation (24 hours at 30°C) in the presence of organic solvent, measured by HPLC

Methanol

30%

59.5

—

18.8

50 mM Tris-HCl, 20 mM G6P, 5U G6PDH

30°C

Benzyl methyl sulfide, O₂

(S)-Benzyl dimethyl sulfoxonium

0.2 mM NADPH

250 rpm

Classical aqueous control (in %)

M446G

Specificity - Enantioselectivity

Enantiomeric excess (toward the S form) after incubation (24 hours at 30°C) in the presence of organic solvent, measured by HPLC

Methanol

50%

59.5

—

6.3

50 mM Tris-HCl, 20 mM G6P, 5U G6PDH

30°C

Benzyl methyl sulfide, O₂

(S)-Benzyl dimethyl sulfoxonium

0.2 mM NADPH

250 rpm

Classical aqueous control (in %)

M446G

Specificity - Enantioselectivity

Enantiomeric excess (toward the S form) after incubation (24 hours at 30°C) in the presence of organic solvent, measured by HPLC

Methanol

70%

59.5

—

0.0

50 mM Tris-HCl, 20 mM G6P, 5U G6PDH

30°C

Benzyl methyl sulfide, O₂

(S)-Benzyl dimethyl sulfoxonium

0.2 mM NADPH

250 rpm

Classical aqueous control (in %)

M446G

Specificity - Enantioselectivity

Enantiomeric excess (toward the S form) after incubation (24 hours at 30°C) in the presence of organic solvent, measured by HPLC

PEG

30%

59.5

—

40.1

50 mM Tris-HCl, 20 mM G6P, 5U G6PDH

30°C

Benzyl methyl sulfide, O₂

(S)-Benzyl dimethyl sulfoxonium

0.2 mM NADPH

250 rpm

Classical aqueous control (in %)

M446G

Specificity - Enantioselectivity

Enantiomeric excess (toward the S form) after incubation (24 hours at 30°C) in the presence of organic solvent, measured by HPLC

Acetonitrile

30%

59.5

—

43.5

50 mM Tris-HCl, 20 mM G6P, 5U G6PDH

30°C

Benzyl methyl sulfide, O₂

(S)-Benzyl dimethyl sulfoxonium

0.2 mM NADPH

250 rpm

Classical aqueous control (in %)

M446G

Specificity - Enantioselectivity

Enantiomeric excess (toward the S form) after incubation (24 hours at 30°C) in the presence of organic solvent, measured by HPLC

Ethanol

30%

59.5

—

31.8

50 mM Tris-HCl, 20 mM G6P, 5U G6PDH

30°C

Benzyl methyl sulfide, O₂

(S)-Benzyl dimethyl sulfoxonium

0.2 mM NADPH

250 rpm

Classical aqueous control (in %)

M446G

Specificity - Enantioselectivity

Enantiomeric excess (toward the S form) after incubation (24 hours at 30°C) in the presence of organic solvent, measured by HPLC

Methanol

79.8

—

84.8

50 mM Tris-HCl, 20 mM G6P, 5U G6PDH

30°C

10 mM methyl phenyl sulfoxide, O₂

(S)-para-nitrophenyl methyl sulfoxide

0.2 mM NADPH

250 rpm

Classical aqueous control (in %)

M446G

Stability - Incubation

Activity after incubation (24 hours at 25°C), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in the presence of organic solvent

PEG

100.0

—

89.0

Incubation: 50 mM Tris-HCl, Assay: 50 mM Tris-HCl

30°C

Benzyl methyl sulfide, O₂

(S)-Benzyl dimethyl sulfoxonium

0.1 mM NADPH

Incubation: 250 rpm

Water-incubated control (in %), 100% value and assay in the presence of organic solvent

M446G

Stability - Incubation

Activity after incubation (24 hours at 25°C), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in the presence of organic solvent

Methanol

100.0

—

84.0

Incubation: 50 mM Tris-HCl, Assay: 50 mM Tris-HCl

30°C

Benzyl methyl sulfide, O₂

(S)-Benzyl dimethyl sulfoxonium

0.1 mM NADPH

Incubation: 250 rpm

Water-incubated control (in %), 100% value and assay in the presence of organic solvent

M446G

Stability - Incubation

Activity after incubation (24 hours at 25°C), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in the presence of organic solvent

Isopropanol

100.0

—

41.0

Incubation: 50 mM Tris-HCl, Assay: 50 mM Tris-HCl

30°C

Benzyl methyl sulfide, O₂

(S)-Benzyl dimethyl sulfoxonium

0.1 mM NADPH

Incubation: 250 rpm

Water-incubated control (in %), 100% value and assay in the presence of organic solvent

M446G

Stability - Incubation

Activity after incubation (24 hours at 25°C), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in the presence of organic solvent

PEG

30%

100.0

—

11.0

Incubation: 50 mM Tris-HCl, Assay: 50 mM Tris-HCl

30°C

Benzyl methyl sulfide, O₂

(S)-Benzyl dimethyl sulfoxonium

0.1 mM NADPH

Incubation: 250 rpm

Water-incubated control (in %), 100% value and assay in the presence of organic solvent

M446G

Stability - Incubation

Activity after incubation (24 hours at 25°C), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in the presence of organic solvent

Methanol

30%

100.0

—

7.0

Incubation: 50 mM Tris-HCl, Assay: 50 mM Tris-HCl

30°C

Benzyl methyl sulfide, O₂

(S)-Benzyl dimethyl sulfoxonium

0.1 mM NADPH

Incubation: 250 rpm

Water-incubated control (in %), 100% value and assay in the presence of organic solvent

M446G

Stability - Incubation

Activity after incubation (24 hours at 25°C), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in the presence of organic solvent

Isopropanol

30%

100.0

—

0.0

Incubation: 50 mM Tris-HCl, Assay: 50 mM Tris-HCl

30°C

Benzyl methyl sulfide, O₂

(S)-Benzyl dimethyl sulfoxonium

0.1 mM NADPH

Incubation: 250 rpm

Water-incubated control (in %), 100% value and assay in the presence of organic solvent

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Mutations in this dataset (1) — Gonzalo de Gonzalo et al. (2012)

M446G

Visualization : Activity — Classical

Gonzalo de Gonzalo et al. (2012)

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

Visualization : Stability — Incubation

Gonzalo de Gonzalo et al. (2012)

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

Visualization : Activity + Stability — Conversion

Gonzalo de Gonzalo et al. (2012)

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

Visualization : Specificity — Enantioselectivity

Gonzalo de Gonzalo et al. (2012)

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

Structure

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Phenylacetone monooxygenase from Thermobifida fusca

Enzyme Description

Sequence

Experimental Data (39 measurements)

Visualization : Activity — Classical

Visualization : Activity + Stability — Incubation

Visualization : Stability — C50

Experimental Data (67 measurements)

Visualization : Activity — Classical

Visualization : Stability — Incubation

Visualization : Activity + Stability — Conversion

Visualization : Specificity — Enantioselectivity

Structure

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Visualization : Stability — C₅₀