Flavin-containing monooxygenase from Nitrincola lacisaponensis
Enzyme Description
Species
Extremophile
Yes
"alkaliphilic extremophilic" organism
EC Number
Sequence
MNTRVAIIGAGPCGLAQLRAFQSAAAKGAAIPELVCFEKQSDWGGMWNYTWRTGVDEHGEPVHGSMYRYLWSNGPKECLEFADYTFDEHFGRPIGSYPPREVLWDYIKGRVEKAGVRDYIRFNTAVRQVSYDETSGLFSVTVQDHSNNRVYTETFDYVVNACGHFSTPNTPYFEGFERFGGRVLHAHDFRDALEFKDKDILVVGSSYSAEDIGSQCYKYGARSITSCYRSAPMGYQWPQNWEEKPLLQRVDSDTAWFADGSHKRIDAIILCTGYQHHFPFLPDSLRLITENRLWPLKLYKGIFWEDNPKFLYLGMQDQWYSFNMFDAQAWYARDVILGRISLPGKAEMQAENQAWREREEKLQTAQEMFEFQGSYIQHLIDATDYPSFDIQAVNETFLAWKHDKYEDIMGYRNKCYRSLMTGTLATPHHTPWLDALDDSLEAYLQETPRLKSAV
Nikola LonΔar et al. (2019)
β Characterization of a thermostable flavin-containing monooxygenase from Nitrincola lacisaponensis (NiFMO)
Applied Microbiology and Biotechnology
Β· doi:10.1007/s00253-018-09579-w β
Β· Activity - Classical Stability - Tm
▼
Database ID
UniProt: A0A063Y6V3 β
Sequence Annotation
Explicit - Provided GenBank Accession Number
Protein Source
Recombinant, host bacterium Escherichia coli NEB10Ξ²
Experimental Data (32 measurements)
32 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 measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in the presence of organic solvent | Ethanol | 5% | 1.05 | 0.5 | 1/s | 50 mM potassium phosphate buffer | 7.5 | 25Β°C | Molecular oxygen, Trimethylamine | H2O , Trimethylamine N-oxide | β | β | Classical aqueous control (in 1/s) |
| Activity - Classical | Activity measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in the presence of organic solvent | Ethanol | 2.5% | 1.05 | 0.3 | 1/s | 50 mM potassium phosphate buffer | 7.5 | 25Β°C | Molecular oxygen, Trimethylamine | H2O , Trimethylamine N-oxide | β | β | Classical aqueous control (in 1/s) |
| Activity - Classical | Activity measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in the presence of organic solvent | Acetonitrile | 10% | 1.05 | 0.6 | 1/s | 50 mM potassium phosphate buffer | 7.5 | 25Β°C | Molecular oxygen, Trimethylamine | H2O , Trimethylamine N-oxide | β | β | Classical aqueous control (in 1/s) |
| Activity - Classical | Activity measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in the presence of organic solvent | Acetonitrile | 7.5% | 1.05 | 0.7 | 1/s | 50 mM potassium phosphate buffer | 7.5 | 25Β°C | Molecular oxygen, Trimethylamine | H2O , Trimethylamine N-oxide | β | β | Classical aqueous control (in 1/s) |
| Activity - Classical | Activity measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in the presence of organic solvent | Acetonitrile | 5% | 1.05 | 0.8 | 1/s | 50 mM potassium phosphate buffer | 7.5 | 25Β°C | Molecular oxygen, Trimethylamine | H2O , Trimethylamine N-oxide | β | β | Classical aqueous control (in 1/s) |
| Activity - Classical | Activity measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in the presence of organic solvent | Acetonitrile | 2.5% | 1.05 | 0.8 | 1/s | 50 mM potassium phosphate buffer | 7.5 | 25Β°C | Molecular oxygen, Trimethylamine | H2O , Trimethylamine N-oxide | β | β | Classical aqueous control (in 1/s) |
| Activity - Classical | Activity measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in the presence of organic solvent | Dimethyl Sulfoxide (DMSO) | 10% | 1.05 | 0.1 | 1/s | 50 mM potassium phosphate buffer | 7.5 | 25Β°C | Molecular oxygen, Trimethylamine | H2O , Trimethylamine N-oxide | β | β | Classical aqueous control (in 1/s) |
| Activity - Classical | Activity measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in the presence of organic solvent | Dimethyl Sulfoxide (DMSO) | 7.5% | 1.05 | 0.2 | 1/s | 50 mM potassium phosphate buffer | 7.5 | 25Β°C | Molecular oxygen, Trimethylamine | H2O , Trimethylamine N-oxide | β | β | Classical aqueous control (in 1/s) |
| Activity - Classical | Activity measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in the presence of organic solvent | Dimethyl Sulfoxide (DMSO) | 5% | 1.05 | 0.2 | 1/s | 50 mM potassium phosphate buffer | 7.5 | 25Β°C | Molecular oxygen, Trimethylamine | H2O , Trimethylamine N-oxide | β | β | Classical aqueous control (in 1/s) |
| Activity - Classical | Activity measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in the presence of organic solvent | Dimethyl Sulfoxide (DMSO) | 2.5% | 1.05 | 0.3 | 1/s | 50 mM potassium phosphate buffer | 7.5 | 25Β°C | Molecular oxygen, Trimethylamine | H2O , Trimethylamine N-oxide | β | β | Classical aqueous control (in 1/s) |
| Activity - Classical | Activity measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in the presence of organic solvent | Methanol | 10% | 1.05 | 0.7 | 1/s | 50 mM potassium phosphate buffer | 7.5 | 25Β°C | Molecular oxygen, Trimethylamine | H2O , Trimethylamine N-oxide | β | β | Classical aqueous control (in 1/s) |
| Activity - Classical | Activity measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in the presence of organic solvent | Methanol | 7.5% | 1.05 | 0.8 | 1/s | 50 mM potassium phosphate buffer | 7.5 | 25Β°C | Molecular oxygen, Trimethylamine | H2O , Trimethylamine N-oxide | β | β | Classical aqueous control (in 1/s) |
| Activity - Classical | Activity measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in the presence of organic solvent | Methanol | 5% | 1.05 | 0.9 | 1/s | 50 mM potassium phosphate buffer | 7.5 | 25Β°C | Molecular oxygen, Trimethylamine | H2O , Trimethylamine N-oxide | β | β | Classical aqueous control (in 1/s) |
| Activity - Classical | Activity measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in the presence of organic solvent | Methanol | 2.5% | 1.05 | 0.8 | 1/s | 50 mM potassium phosphate buffer | 7.5 | 25Β°C | Molecular oxygen, Trimethylamine | H2O , Trimethylamine N-oxide | β | β | Classical aqueous control (in 1/s) |
| Activity - Classical | Activity measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in the presence of organic solvent | Ethanol | 10% | 1.05 | 0.4 | 1/s | 50 mM potassium phosphate buffer | 7.5 | 25Β°C | Molecular oxygen, Trimethylamine | H2O , Trimethylamine N-oxide | β | β | Classical aqueous control (in 1/s) |
| Activity - Classical | Activity measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in the presence of organic solvent | Ethanol | 7.5% | 1.05 | 0.5 | 1/s | 50 mM potassium phosphate buffer | 7.5 | 25Β°C | Molecular oxygen, Trimethylamine | H2O , Trimethylamine N-oxide | β | β | Classical aqueous control (in 1/s) |
| Stability - Tm | Tm measured by ThermoFAD (detection of FAD fluorescence change upon protein unfolding) in the presence of organic solvent | Acetonitrile | 10% | 51.0 | 39.3 | Β°C | 50 mM potassium phosphate buffer | 7.5 | β | β | β | β | β | Classical aqueous control (in Β°C) |
| Stability - Tm | Tm measured by ThermoFAD (detection of FAD fluorescence change upon protein unfolding) in the presence of organic solvent | Acetonitrile | 7.5% | 51.0 | 41.3 | Β°C | 50 mM potassium phosphate buffer | 7.5 | β | β | β | β | β | Classical aqueous control (in Β°C) |
| Stability - Tm | Tm measured by ThermoFAD (detection of FAD fluorescence change upon protein unfolding) in the presence of organic solvent | Acetonitrile | 5% | 51.0 | 44.5 | Β°C | 50 mM potassium phosphate buffer | 7.5 | β | β | β | β | β | Classical aqueous control (in Β°C) |
| Stability - Tm | Tm measured by ThermoFAD (detection of FAD fluorescence change upon protein unfolding) in the presence of organic solvent | Acetonitrile | 2.5% | 51.0 | 45.5 | Β°C | 50 mM potassium phosphate buffer | 7.5 | β | β | β | β | β | Classical aqueous control (in Β°C) |
Visualization : Activity β Classical
One bar per measurement. Colour = solvent, shade = solvent volume.
Visualization : Stability β Tm
One bar per measurement. Colour = solvent, shade = solvent volume.