Cytochrome P450 CYP119 from Sulfolobus acidocaldarius
Enzyme Description
Species
Extremophile
Yes
thermoacidophilic organism
EC Number
Sequence
MYDWFSEMRKKDPVYYDGNIWQVFSYRYTKEVLNNFSKFSSDLTGYHERLEDLRNGKIRFDIPTRYTMLTSDPPLHDELRSMSADIFSPQKLQTLETFIRETTRSLLDSIDPREDDIVKKLAVPLPIIVISKILGLPIEDKEKFKEWSDLVAFRLGKPGEIFELGKKYLELIGYVKDHLNSGTEVVSRVVNSNLSDIEKLGYIILLLIAGNETTTNLISNSVIDFTRFNLWQRIREENLYLKAIEEALRYSPPVMRTVRKTKERVKLGDQTIEEGEYVRVWIASANRDEEVFHDGEKFIPDRNPNPHLSFGSGIHLCLGAPLARLEARIAIEEFSKRFRHIEILDTEKVPNEVLNGYKRLVVRLKSNE
Kersten S. Rabe et al. (2010)
β Peroxidase activity of bacterial cytochrome P450 enzymes: Modulation by fatty acids and organic solvents
Biotechnology Journal
Β· doi:10.1002/biot.201000028 β
Β· Activity - Classical
▼
Database ID
UniProt: Q55080 β
Sequence Annotation
Inferred
Protein Source
Recombinant, host bacterium Escherichia coli BL21 (DE3)
Experimental Data (4 measurements)
4 measurements
| 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 nmol substrate per nmol enzyme per second) measured by fluorescence spectrophotometry (resorufin fluorescence measurement) in the presence of organic solvent | Methanol | 10% | 0.0005 | 0.0025 | nmol substrate per nmol enzyme per second | 50 mM potassium phosphate buffer | 7 | 25Β°C | 1 Β΅M Amplex red, 1 mM HβOβ (Hydrogen Peroxide) | H2O , Resorufin | β | β | Classical aqueous control (in nmol substrate per nmol enzyme per second)) | Here peroxidase activity measured, hence the EC number choice |
| Activity - Classical | Activity (in nmol substrate per nmol enzyme per second) measured by fluorescence spectrophotometry (resorufin fluorescence measurement) in the presence of organic solvent | Ethanol | 10% | 0.0005 | 0.0021 | nmol substrate per nmol enzyme per second | 50 mM potassium phosphate buffer | 7 | 25Β°C | 1 Β΅M Amplex red, 1 mM HβOβ (Hydrogen Peroxide) | H2O , Resorufin | β | β | Classical aqueous control (in nmol substrate per nmol enzyme per second)) | Here peroxidase activity measured, hence the EC number choice |
| Activity - Classical | Activity (in nmol substrate per nmol enzyme per second) measured by fluorescence spectrophotometry (resorufin fluorescence measurement) in the presence of organic solvent | Isopropanol | 10% | 0.0005 | 0.0034 | nmol substrate per nmol enzyme per second | 50 mM potassium phosphate buffer | 7 | 25Β°C | 1 Β΅M Amplex red, 1 mM HβOβ (Hydrogen Peroxide) | H2O , Resorufin | β | β | Classical aqueous control (in nmol substrate per nmol enzyme per second)) | Here peroxidase activity measured, hence the EC number choice |
| Activity - Classical | Activity (in nmol substrate per nmol enzyme per second) measured by fluorescence spectrophotometry (resorufin fluorescence measurement) in the presence of organic solvent | Acetonitrile | 10% | 0.0005 | 0.0022 | nmol substrate per nmol enzyme per second | 50 mM potassium phosphate buffer | 7 | 25Β°C | 1 Β΅M Amplex red, 1 mM HβOβ (Hydrogen Peroxide) | H2O , Resorufin | β | β | Classical aqueous control (in nmol substrate per nmol enzyme per second)) | Here peroxidase activity measured, hence the EC number choice |
Visualization : Activity β Classical
Kersten S. Rabe et al. (2010)
One bar per measurement. Colour = solvent, shade = solvent volume.
Tugce Sakalli et al. (2021)
β Functional characterization of a novel CYP119 variant to explore its biocatalytic potential
Biotechnology and Applied Biochemistry
Β· doi:10.1002/bab.2243 β
Β· Activity - Classical
▼
Database ID
PDB: 1F4T β
Sequence Annotation
Explicit - Provided PDB Accession Number
Protein Source
Recombinant, host bacterium Escherichia coli BL21
Experimental Data (18 measurements)
18 measurements
| Mutation | Property | Assay | Solvent | Solvent Volume | Aqueous Reference | WT Reference | Measured Value | Units | Solution | pH | Temperature | Substrate(s) | Product(s) | Cofactor(s) | Shaking | Comments |
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| WT | Activity - Classical | Activity measured by fluorescence spectrophotometry (resorufin fluorescence measurement, excited at 545 nm and emission at 595 nm) in the presence of organic solvent | Methanol | 5% | 100.0 | β | 87.7 | % | 50 mM potassium phosphate buffer | 7.4 | Room temperature | 10 Β΅M Amplex red, HβOβ (Hydrogen Peroxide) | H2O , Resorufin | β | β | Classical aqueous control (in %) |
| WT | Activity - Classical | Activity measured by fluorescence spectrophotometry (resorufin fluorescence measurement, excited at 545 nm and emission at 595 nm) in the presence of organic solvent | Acetonitrile | 7.5% | 100.0 | β | 58.9 | % | 50 mM potassium phosphate buffer | 7.4 | Room temperature | 10 Β΅M Amplex red, HβOβ (Hydrogen Peroxide) | H2O , Resorufin | β | β | Classical aqueous control (in %) |
| WT | Activity - Classical | Activity measured by fluorescence spectrophotometry (resorufin fluorescence measurement, excited at 545 nm and emission at 595 nm) in the presence of organic solvent | Methanol | 7.5% | 100.0 | β | 65.0 | % | 50 mM potassium phosphate buffer | 7.4 | Room temperature | 10 Β΅M Amplex red, HβOβ (Hydrogen Peroxide) | H2O , Resorufin | β | β | Classical aqueous control (in %) |
| WT | Activity - Classical | Activity measured by fluorescence spectrophotometry (resorufin fluorescence measurement, excited at 545 nm and emission at 595 nm) in the presence of organic solvent | Isopropanol | 2.5% | 100.0 | β | 87.1 | % | 50 mM potassium phosphate buffer | 7.4 | Room temperature | 10 Β΅M Amplex red, HβOβ (Hydrogen Peroxide) | H2O , Resorufin | β | β | Classical aqueous control (in %) |
| WT | Activity - Classical | Activity measured by fluorescence spectrophotometry (resorufin fluorescence measurement, excited at 545 nm and emission at 595 nm) in the presence of organic solvent | Acetonitrile | 5% | 100.0 | β | 76.7 | % | 50 mM potassium phosphate buffer | 7.4 | Room temperature | 10 Β΅M Amplex red, HβOβ (Hydrogen Peroxide) | H2O , Resorufin | β | β | Classical aqueous control (in %) |
| WT | Activity - Classical | Activity measured by fluorescence spectrophotometry (resorufin fluorescence measurement, excited at 545 nm and emission at 595 nm) in the presence of organic solvent | Isopropanol | 5% | 100.0 | β | 63.9 | % | 50 mM potassium phosphate buffer | 7.4 | Room temperature | 10 Β΅M Amplex red, HβOβ (Hydrogen Peroxide) | H2O , Resorufin | β | β | Classical aqueous control (in %) |
| WT | Activity - Classical | Activity measured by fluorescence spectrophotometry (resorufin fluorescence measurement, excited at 545 nm and emission at 595 nm) in the presence of organic solvent | Methanol | 2.5% | 100.0 | β | 90.2 | % | 50 mM potassium phosphate buffer | 7.4 | Room temperature | 10 Β΅M Amplex red, HβOβ (Hydrogen Peroxide) | H2O , Resorufin | β | β | Classical aqueous control (in %) |
| WT | Activity - Classical | Activity measured by fluorescence spectrophotometry (resorufin fluorescence measurement, excited at 545 nm and emission at 595 nm) in the presence of organic solvent | Isopropanol | 7.5% | 100.0 | β | 49.7 | % | 50 mM potassium phosphate buffer | 7.4 | Room temperature | 10 Β΅M Amplex red, HβOβ (Hydrogen Peroxide) | H2O , Resorufin | β | β | Classical aqueous control (in %) |
| WT | Activity - Classical | Activity measured by fluorescence spectrophotometry (resorufin fluorescence measurement, excited at 545 nm and emission at 595 nm) in the presence of organic solvent | Acetonitrile | 2.5% | 100.0 | β | 92.0 | % | 50 mM potassium phosphate buffer | 7.4 | Room temperature | 10 Β΅M Amplex red, HβOβ (Hydrogen Peroxide) | H2O , Resorufin | β | β | Classical aqueous control (in %) |
| T213R/T214I | Activity - Classical | Activity measured by fluorescence spectrophotometry (resorufin fluorescence measurement, excited at 545 nm and emission at 595 nm) in the presence of organic solvent | Isopropanol | 5% | 100.0 | 63.9 | 46.4 | % | 50 mM potassium phosphate buffer | 7.4 | Room temperature | 10 Β΅M Amplex red, HβOβ (Hydrogen Peroxide) | H2O , Resorufin | β | β | Classical aqueous control (in %) |
| T213R/T214I | Activity - Classical | Activity measured by fluorescence spectrophotometry (resorufin fluorescence measurement, excited at 545 nm and emission at 595 nm) in the presence of organic solvent | Acetonitrile | 7.5% | 100.0 | 58.9 | 73.6 | % | 50 mM potassium phosphate buffer | 7.4 | Room temperature | 10 Β΅M Amplex red, HβOβ (Hydrogen Peroxide) | H2O , Resorufin | β | β | Classical aqueous control (in %) |
| T213R/T214I | Activity - Classical | Activity measured by fluorescence spectrophotometry (resorufin fluorescence measurement, excited at 545 nm and emission at 595 nm) in the presence of organic solvent | Acetonitrile | 5% | 100.0 | 76.7 | 79.1 | % | 50 mM potassium phosphate buffer | 7.4 | Room temperature | 10 Β΅M Amplex red, HβOβ (Hydrogen Peroxide) | H2O , Resorufin | β | β | Classical aqueous control (in %) |
| T213R/T214I | Activity - Classical | Activity measured by fluorescence spectrophotometry (resorufin fluorescence measurement, excited at 545 nm and emission at 595 nm) in the presence of organic solvent | Acetonitrile | 2.5% | 100.0 | 92.0 | 90.2 | % | 50 mM potassium phosphate buffer | 7.4 | Room temperature | 10 Β΅M Amplex red, HβOβ (Hydrogen Peroxide) | H2O , Resorufin | β | β | Classical aqueous control (in %) |
| T213R/T214I | Activity - Classical | Activity measured by fluorescence spectrophotometry (resorufin fluorescence measurement, excited at 545 nm and emission at 595 nm) in the presence of organic solvent | Isopropanol | 7.5% | 100.0 | 49.7 | 49.0 | % | 50 mM potassium phosphate buffer | 7.4 | Room temperature | 10 Β΅M Amplex red, HβOβ (Hydrogen Peroxide) | H2O , Resorufin | β | β | Classical aqueous control (in %) |
| T213R/T214I | Activity - Classical | Activity measured by fluorescence spectrophotometry (resorufin fluorescence measurement, excited at 545 nm and emission at 595 nm) in the presence of organic solvent | Isopropanol | 2.5% | 100.0 | 87.1 | 59.4 | % | 50 mM potassium phosphate buffer | 7.4 | Room temperature | 10 Β΅M Amplex red, HβOβ (Hydrogen Peroxide) | H2O , Resorufin | β | β | Classical aqueous control (in %) |
| T213R/T214I | Activity - Classical | Activity measured by fluorescence spectrophotometry (resorufin fluorescence measurement, excited at 545 nm and emission at 595 nm) in the presence of organic solvent | Methanol | 7.5% | 100.0 | 65.0 | 77.9 | % | 50 mM potassium phosphate buffer | 7.4 | Room temperature | 10 Β΅M Amplex red, HβOβ (Hydrogen Peroxide) | H2O , Resorufin | β | β | Classical aqueous control (in %) |
| T213R/T214I | Activity - Classical | Activity measured by fluorescence spectrophotometry (resorufin fluorescence measurement, excited at 545 nm and emission at 595 nm) in the presence of organic solvent | Methanol | 5% | 100.0 | 87.7 | 77.3 | % | 50 mM potassium phosphate buffer | 7.4 | Room temperature | 10 Β΅M Amplex red, HβOβ (Hydrogen Peroxide) | H2O , Resorufin | β | β | Classical aqueous control (in %) |
| T213R/T214I | Activity - Classical | Activity measured by fluorescence spectrophotometry (resorufin fluorescence measurement, excited at 545 nm and emission at 595 nm) in the presence of organic solvent | Methanol | 2.5% | 100.0 | 90.2 | 91.4 | % | 50 mM potassium phosphate buffer | 7.4 | Room temperature | 10 Β΅M Amplex red, HβOβ (Hydrogen Peroxide) | H2O , Resorufin | β | β | Classical aqueous control (in %) |
Mutations in this dataset (2) β Tugce Sakalli et al. (2021)
WT
T213R/T214I
Visualization : Activity β Classical
Tugce Sakalli et al. (2021)
One bar per measurement. Colour = solvent, shade = solvent volume. β β β 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.