Phenylacetone monooxygenase from Thermobifida fusca
Enzyme Description
Species
Extremophile
Yes
thermophilic organism
EC Number
1.14.13.92 β,
1.14.13.92 β (sulfoxidation)
Sequence
MAGQTTVDSRRQPPEEVDVLVVGAGFSGLYALYRLRELGRSVHVIETAGDVGGVWYWNRYPGARCDIESIEYCYSFSEEVLQEWNWTERYASQPEILRYINFVADKFDLRSGITFHTTVTAAAFDEATNTWTVDTNHGDRIRARYLIMASGQLSVPQLPNFPGLKDFAGNLYHTGNWPHEPVDFSGQRVGVIGTGSSGIQVSPQIAKQAAELFVFQRTPHFAVPARNAPLDPEFLADLKKRYAEFREESRNTPGGTHRYQGPKSALEVSDEELVETLERYWQEGGPDILAAYRDILRDRDANERVAEFIRNKIRNTVRDPEVAERLVPKGYPFGTKRLILEIDYYEMFNRDNVHLVDTLSAPIETITPRGVRTSEREYELDSLVLATGFDALTGALFKIDIRGVGNVALKEKWAAGPRTYLGLSTAGFPNLFFIAGPGSPSALSNMLVSIEQHVEWVTDHIAYMFKNGLTRSEAVLEKEDEWVEHVNEIADETLYPMTASWYTGANVPGKPRVFMLYVGGFHRYRQICDEVAAKGYEGFVLT
Francesco Secundo et al. (2010)
β Effects of water miscible organic solvents on the activity and conformation of the baeyerβvilliger monooxygenases from Thermobifida fusca and Acinetobacter calcoaceticus : A comparative study
Biotechnology and Bioengineering
Β· doi:10.1002/bit.22963 β
Β· Activity - Classical Stability - C50 Activity + Stability - Incubation
▼
Database ID
UniProt: Q47PU3 β
Sequence Annotation
Inferred - from protein name
Protein Source
Recombinant, host bacterium Escherichia coli BL21
Experimental Data (39 measurements)
39 measurements β 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 | 25Β°C | Molecular oxygen (O2), Thioanisole | H2O , methyl phenyl sulfoxide | 0.1 mM NADPH | β | 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 | 25Β°C | Molecular oxygen (O2), Thioanisole | H2O , methyl phenyl sulfoxide | 0.1 mM NADPH | β | 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 | 25Β°C | Molecular oxygen (O2), Thioanisole | H2O , methyl phenyl sulfoxide | 0.1 mM NADPH | β | 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 | 25Β°C | Molecular oxygen (O2), Thioanisole | H2O , methyl phenyl sulfoxide | 0.1 mM NADPH | β | 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 | 25Β°C | Molecular oxygen (O2), Thioanisole | H2O , methyl phenyl sulfoxide | 0.1 mM NADPH | β | 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 | 25Β°C | Molecular oxygen (O2), Thioanisole | H2O , methyl phenyl sulfoxide | 0.1 mM NADPH | β | 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 | 25Β°C | Molecular oxygen (O2), Thioanisole | H2O , methyl phenyl sulfoxide | 0.1 mM NADPH | β | 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 | 25Β°C | Molecular oxygen (O2), Thioanisole | H2O , methyl phenyl sulfoxide | 0.1 mM NADPH | β | 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 | 25Β°C | Molecular oxygen (O2), Thioanisole | H2O , methyl phenyl sulfoxide | 0.1 mM NADPH | β | 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 | 25Β°C | Molecular oxygen (O2), Thioanisole | H2O , methyl phenyl sulfoxide | 0.1 mM NADPH | β | 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 | 25Β°C | Molecular oxygen (O2), Thioanisole | H2O , methyl phenyl sulfoxide | 0.1 mM NADPH | β | 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 | 25Β°C | Molecular oxygen (O2), Thioanisole | H2O , methyl phenyl sulfoxide | 0.1 mM NADPH | β | 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 | 25Β°C | Molecular oxygen (O2), Thioanisole | H2O , methyl phenyl sulfoxide | 0.1 mM NADPH | β | 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 | 25Β°C | Molecular oxygen (O2), Thioanisole | H2O , methyl phenyl sulfoxide | 0.1 mM NADPH | β | 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 | 25Β°C | Molecular oxygen (O2), Thioanisole | H2O , methyl phenyl sulfoxide | 0.1 mM NADPH | β | 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 | 25Β°C | Molecular oxygen (O2), Thioanisole | H2O , methyl phenyl sulfoxide | 0.1 mM NADPH | β | 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 | 25Β°C | Molecular oxygen (O2), Thioanisole | H2O , methyl phenyl sulfoxide | 0.1 mM NADPH | β | 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 | 25Β°C | Molecular oxygen (O2), Thioanisole | H2O , methyl phenyl sulfoxide | 0.1 mM NADPH | β | 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 | 25Β°C | Molecular oxygen (O2), Thioanisole | H2O , methyl phenyl sulfoxide | 0.1 mM NADPH | β | 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 | 25Β°C | Molecular oxygen (O2), Thioanisole | H2O , methyl phenyl sulfoxide | 0.1 mM NADPH | β | Classical aqueous control (in U/mg) |
Visualization : Activity β Classical
Francesco Secundo et al. (2010)
One bar per measurement. Colour = solvent, shade = solvent volume.
Visualization : Activity + Stability β Incubation
Francesco Secundo et al. (2010)
One bar per measurement. Colour = solvent, shade = solvent volume.
Visualization : Stability β C50
Francesco Secundo et al. (2010)
One bar per measurement. Colour = solvent, shade = temperature. Hover for details.
Gonzalo de Gonzalo et al. (2012)
β Improvement of the biocatalytic properties of one phenylacetone monooxygenase mutant in hydrophilic organic solvents
Enzyme and Microbial Technology
Β· doi:10.1016/j.enzmictec.2011.09.006 β
Β· Activity - Classical Stability - Incubation Activity + Stability - Conversion Specificity - Enantioselectivity
▼
Database ID
UniProt: Q47PU3 β
Sequence Annotation
Inferred - from protein name (1 mutation)
Protein Source
Recombinant, host bacterium Escherichia coli BL21
Experimental Data (67 measurements)
67 measurements β page 3 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 30Β°C) in the presence of organic solvent, measured by HPLC | Methanol | 5% | 50.3 | β | 51.2 | % | 50 mM Tris-HCl, 20 mM G6P, 5U G6PDH | 9 | 30Β°C | 10 mM Cyclohexyl methyl sulfide, Oβ | (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 30Β°C) in the presence of organic solvent, measured by HPLC | Methanol | 5% | 30.3 | β | 36.7 | % | 50 mM Tris-HCl, 20 mM G6P, 5U G6PDH | 9 | 30Β°C | 10 mM Cyclohexyl ethyl sulfide, Oβ | (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 30Β°C) in the presence of organic solvent, measured by HPLC | Methanol | 5% | 0.0 | β | 20.8 | % | 50 mM Tris-HCl, 20 mM G6P, 5U G6PDH | 9 | 30Β°C | 10 mM Cyclohexyl propyl sulfide, Oβ | (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 30Β°C) in the presence of organic solvent, measured by HPLC | Methanol | 5% | 89.8 | β | 93.3 | % | 50 mM Tris-HCl, 20 mM G6P, 5U G6PDH | 9 | 30Β°C | Oβ, 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 30Β°C) in the presence of organic solvent, measured by HPLC | PEG | 10% | 59.5 | β | 63.3 | % | 50 mM Tris-HCl, 20 mM G6P, 5U G6PDH | 9 | 30Β°C | Benzyl methyl sulfide, Oβ | (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 30Β°C) in the presence of organic solvent, measured by HPLC | Methanol | 10% | 59.5 | β | 59.5 | % | 50 mM Tris-HCl, 20 mM G6P, 5U G6PDH | 9 | 30Β°C | Benzyl methyl sulfide, Oβ | (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 30Β°C) in the presence of organic solvent, measured by HPLC | Acetonitrile | 10% | 59.5 | β | 57.3 | % | 50 mM Tris-HCl, 20 mM G6P, 5U G6PDH | 9 | 30Β°C | Benzyl methyl sulfide, Oβ | (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 30Β°C) in the presence of organic solvent, measured by HPLC | Ethanol | 10% | 59.5 | β | 61.3 | % | 50 mM Tris-HCl, 20 mM G6P, 5U G6PDH | 9 | 30Β°C | Benzyl methyl sulfide, Oβ | (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 30Β°C) in the presence of organic solvent, measured by HPLC | Isopropanol | 10% | 59.5 | β | 64.8 | % | 50 mM Tris-HCl, 20 mM G6P, 5U G6PDH | 9 | 30Β°C | Benzyl methyl sulfide, Oβ | (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 30Β°C) in the presence of organic solvent, measured by HPLC | Ethyl Acetate | 10% | 59.5 | β | 58.8 | % | 50 mM Tris-HCl, 20 mM G6P, 5U G6PDH | 9 | 30Β°C | Benzyl methyl sulfide, Oβ | (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 30Β°C) in the presence of organic solvent, measured by HPLC | Diisopropyl Ether | 10% | 59.5 | β | 56.0 | % | 50 mM Tris-HCl, 20 mM G6P, 5U G6PDH | 9 | 30Β°C | Benzyl methyl sulfide, Oβ | (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 30Β°C) in the presence of organic solvent, measured by HPLC | Toluene | 10% | 59.5 | β | 59.0 | % | 50 mM Tris-HCl, 20 mM G6P, 5U G6PDH | 9 | 30Β°C | Benzyl methyl sulfide, Oβ | (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 30Β°C) in the presence of organic solvent, measured by HPLC | n-Hexane | 10% | 59.5 | β | 58.8 | % | 50 mM Tris-HCl, 20 mM G6P, 5U G6PDH | 9 | 30Β°C | Benzyl methyl sulfide, Oβ | (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 30Β°C) in the presence of organic solvent, measured by HPLC | Methanol | 5% | 59.5 | β | 62.5 | % | 50 mM Tris-HCl, 20 mM G6P, 5U G6PDH | 9 | 30Β°C | Benzyl methyl sulfide, Oβ | (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 30Β°C) in the presence of organic solvent, measured by HPLC | Methanol | 30% | 59.5 | β | 18.8 | % | 50 mM Tris-HCl, 20 mM G6P, 5U G6PDH | 9 | 30Β°C | Benzyl methyl sulfide, Oβ | (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 30Β°C) in the presence of organic solvent, measured by HPLC | Methanol | 50% | 59.5 | β | 6.3 | % | 50 mM Tris-HCl, 20 mM G6P, 5U G6PDH | 9 | 30Β°C | Benzyl methyl sulfide, Oβ | (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 30Β°C) in the presence of organic solvent, measured by HPLC | Methanol | 70% | 59.5 | β | 0.0 | % | 50 mM Tris-HCl, 20 mM G6P, 5U G6PDH | 9 | 30Β°C | Benzyl methyl sulfide, Oβ | (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 30Β°C) in the presence of organic solvent, measured by HPLC | PEG | 30% | 59.5 | β | 40.1 | % | 50 mM Tris-HCl, 20 mM G6P, 5U G6PDH | 9 | 30Β°C | Benzyl methyl sulfide, Oβ | (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 30Β°C) in the presence of organic solvent, measured by HPLC | Acetonitrile | 30% | 59.5 | β | 43.5 | % | 50 mM Tris-HCl, 20 mM G6P, 5U G6PDH | 9 | 30Β°C | Benzyl methyl sulfide, Oβ | (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 30Β°C) in the presence of organic solvent, measured by HPLC | Ethanol | 30% | 59.5 | β | 31.8 | % | 50 mM Tris-HCl, 20 mM G6P, 5U G6PDH | 9 | 30Β°C | Benzyl methyl sulfide, Oβ | (S)-Benzyl dimethyl sulfoxonium | 0.2 mM NADPH | 250 rpm | Classical aqueous control (in %) |
Mutations in this dataset (1) β Gonzalo de Gonzalo et al. (2012)
M446G
Visualization : Activity β Classical
Gonzalo de Gonzalo et al. (2012)
One bar per measurement. Colour = solvent, shade = solvent volume.
Visualization : Stability β Incubation
Gonzalo de Gonzalo et al. (2012)
One bar per measurement. Colour = solvent, shade = solvent volume.
Visualization : Activity + Stability β Conversion
Gonzalo de Gonzalo et al. (2012)
One bar per measurement. Colour = solvent, shade = solvent volume.
Visualization : Specificity β Enantioselectivity
Gonzalo de Gonzalo et al. (2012)
One bar per measurement. Colour = solvent, shade = solvent volume.