Phthalate-degrading esterase from a soil metagenomic library
Enzyme Description
Sequence
MASPQLQMALDAFKTMGEKMAQAGNDVKALRAVMEEMSGFPSAGETKCPTVNAGGVPAEWISGPGAAADDRVILLYHGGGYVMGSIATHRETVARLSKASGARGIALDYRLAPEHPPAAVDDATAAYRWLLSQNIKPAHIVIAGDSAGGGLTLATLIARDAKVPLPAAGVCISPWTDMEGAGESMTTRAKADPVVQKQGLLGMAQLYLGGKDPKSPLAAPLHANLAGLPPILIQVGDAETLLDDSIRVAEKAKKAGVKVDLEWPEMPHVWHLFAPFLPEGQQAIDKIGKYVRQITA
Jiarong Qiu et al. (2021)
β Enhancing the activity and thermal stability of a phthalate-degrading hydrolase by random mutagenesis
Ecotoxicology and Environmental Safety
Β· doi:10.1016/j.ecoenv.2020.111795 β
Β· Activity - Classical
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Database ID
β
Sequence Annotation
Explicit - Provided GenBank Accession Number
Protein Source
Recombinant, host bacterium Escherichia coli BL21 (DE3)
Experimental Data (20 measurements)
20 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 HPLC ( product formation measurement) | Ethanol | 50% | 100 | β | 44 | % | 100 mM Tris-HCl buffer | 8 | 40Β°C | 1 mM Dibutyl phthalate | 1-Butanol , Monobutyl phtalate | β | β | Classical aqueous control (in %) |
| WT | Activity - Classical | Activity measured by HPLC ( product formation measurement) | Dimethyl Sulfoxide (DMSO) | 50% | 100 | β | 58 | % | 100 mM Tris-HCl buffer | 8 | 40Β°C | 1 mM Dibutyl phthalate | 1-Butanol , Monobutyl phtalate | β | β | Classical aqueous control (in %) |
| WT | Activity - Classical | Activity measured by HPLC ( product formation measurement) | Isopropanol | 50% | 100 | β | 53 | % | 100 mM Tris-HCl buffer | 8 | 40Β°C | 1 mM Dibutyl phthalate | 1-Butanol , Monobutyl phtalate | β | β | Classical aqueous control (in %) |
| WT | Activity - Classical | Activity measured by HPLC ( product formation measurement) | Methanol | 50% | 100 | β | 24 | % | 100 mM Tris-HCl buffer | 8 | 40Β°C | 1 mM Dibutyl phthalate | 1-Butanol , Monobutyl phtalate | β | β | Classical aqueous control (in %) |
| WT | Activity - Classical | Activity measured by HPLC ( product formation measurement) | Acetone | 50% | 100 | β | 40 | % | 100 mM Tris-HCl buffer | 8 | 40Β°C | 1 mM Dibutyl phthalate | 1-Butanol , Monobutyl phtalate | β | β | Classical aqueous control (in %) |
| T91M/K163N/L274Q | Activity - Classical | Activity measured by HPLC ( product formation measurement) | Isopropanol | 50% | 100 | 53 | 60 | % | 100 mM Tris-HCl buffer | 8 | 40Β°C | 1 mM Dibutyl phthalate | 1-Butanol , Monobutyl phtalate | β | β | Classical aqueous control (in %) |
| A67V/T91M/V249I | Activity - Classical | Activity measured by HPLC ( product formation measurement) | Dimethyl Sulfoxide (DMSO) | 50% | 100 | 58 | 66 | % | 100 mM Tris-HCl buffer | 8 | 40Β°C | 1 mM Dibutyl phthalate | 1-Butanol , Monobutyl phtalate | β | β | Classical aqueous control (in %) |
| T91M/K163N/L274Q | Activity - Classical | Activity measured by HPLC ( product formation measurement) | Dimethyl Sulfoxide (DMSO) | 50% | 100 | 58 | 60 | % | 100 mM Tris-HCl buffer | 8 | 40Β°C | 1 mM Dibutyl phthalate | 1-Butanol , Monobutyl phtalate | β | β | Classical aqueous control (in %) |
| V75L/K288E | Activity - Classical | Activity measured by HPLC ( product formation measurement) | Dimethyl Sulfoxide (DMSO) | 50% | 100 | 58 | 54 | % | 100 mM Tris-HCl buffer | 8 | 40Β°C | 1 mM Dibutyl phthalate | 1-Butanol , Monobutyl phtalate | β | β | Classical aqueous control (in %) |
| A67V/T91M/V249I | Activity - Classical | Activity measured by HPLC ( product formation measurement) | Acetone | 50% | 100 | 40 | 55 | % | 100 mM Tris-HCl buffer | 8 | 40Β°C | 1 mM Dibutyl phthalate | 1-Butanol , Monobutyl phtalate | β | β | Classical aqueous control (in %) |
| T91M/K163N/L274Q | Activity - Classical | Activity measured by HPLC ( product formation measurement) | Acetone | 50% | 100 | 40 | 48 | % | 100 mM Tris-HCl buffer | 8 | 40Β°C | 1 mM Dibutyl phthalate | 1-Butanol , Monobutyl phtalate | β | β | Classical aqueous control (in %) |
| V75L/K288E | Activity - Classical | Activity measured by HPLC ( product formation measurement) | Acetone | 50% | 100 | 40 | 48 | % | 100 mM Tris-HCl buffer | 8 | 40Β°C | 1 mM Dibutyl phthalate | 1-Butanol , Monobutyl phtalate | β | β | Classical aqueous control (in %) |
| A67V/T91M/V249I | Activity - Classical | Activity measured by HPLC ( product formation measurement) | Isopropanol | 50% | 100 | 53 | 58 | % | 100 mM Tris-HCl buffer | 8 | 40Β°C | 1 mM Dibutyl phthalate | 1-Butanol , Monobutyl phtalate | β | β | Classical aqueous control (in %) |
| V75L/K288E | Activity - Classical | Activity measured by HPLC ( product formation measurement) | Isopropanol | 50% | 100 | 53 | 51 | % | 100 mM Tris-HCl buffer | 8 | 40Β°C | 1 mM Dibutyl phthalate | 1-Butanol , Monobutyl phtalate | β | β | Classical aqueous control (in %) |
| A67V/T91M/V249I | Activity - Classical | Activity measured by HPLC ( product formation measurement) | Ethanol | 50% | 100 | 44 | 59 | % | 100 mM Tris-HCl buffer | 8 | 40Β°C | 1 mM Dibutyl phthalate | 1-Butanol , Monobutyl phtalate | β | β | Classical aqueous control (in %) |
| T91M/K163N/L274Q | Activity - Classical | Activity measured by HPLC ( product formation measurement) | Ethanol | 50% | 100 | 44 | 46 | % | 100 mM Tris-HCl buffer | 8 | 40Β°C | 1 mM Dibutyl phthalate | 1-Butanol , Monobutyl phtalate | β | β | Classical aqueous control (in %) |
| V75L/K288E | Activity - Classical | Activity measured by HPLC ( product formation measurement) | Ethanol | 50% | 100 | 44 | 43 | % | 100 mM Tris-HCl buffer | 8 | 40Β°C | 1 mM Dibutyl phthalate | 1-Butanol , Monobutyl phtalate | β | β | Classical aqueous control (in %) |
| A67V/T91M/V249I | Activity - Classical | Activity measured by HPLC ( product formation measurement) | Methanol | 50% | 100 | 24 | 52 | % | 100 mM Tris-HCl buffer | 8 | 40Β°C | 1 mM Dibutyl phthalate | 1-Butanol , Monobutyl phtalate | β | β | Classical aqueous control (in %) |
| T91M/K163N/L274Q | Activity - Classical | Activity measured by HPLC ( product formation measurement) | Methanol | 50% | 100 | 24 | 43 | % | 100 mM Tris-HCl buffer | 8 | 40Β°C | 1 mM Dibutyl phthalate | 1-Butanol , Monobutyl phtalate | β | β | Classical aqueous control (in %) |
| V75L/K288E | Activity - Classical | Activity measured by HPLC ( product formation measurement) | Methanol | 50% | 100 | 24 | 28 | % | 100 mM Tris-HCl buffer | 8 | 40Β°C | 1 mM Dibutyl phthalate | 1-Butanol , Monobutyl phtalate | β | β | Classical aqueous control (in %) |
Mutations in this dataset (4)
WT
A67V/T91M/V249I
V75L/K288E
T91M/K163N/L274Q
Visualization : Activity β Classical
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.