4-oxalocrotonate tautomerase from Pseudomonas putida

Enzyme Description

Extremophile
No
EC Number
5.3.2.6 β†— (natural activity but non-natural Michaelis addition activity evaluated)

Sequence

Length: 62 amino acids
PIAQIHILEGRSDEQKETLIREVSEAISRSLDAPLTSVRVIITEMAKGHFGIGGELASKVRR
Chao Guo et al. (2020) β€” Tuning Enzyme Activity for Nonaqueous Solvents: Engineering an Enantioselective "Michaelase" for Catalysis in High Concentrations of Ethanol
ChemBioChem  Β· doi:10.1002/cbic.201900721 β†—  Β· Activity - Classical
1104 measurements
Database ID
UniProt: Q01468 β†—
Sequence Annotation
Explicit - Provided (first residue from UniProt sequence removed from mature protein, 1 mutation)
Protein Source
Recombinant, host bacterium Escherichia coli BL21 (DE3)

Experimental Data (1104 measurements)

1104 measurements β€” page 1 of 56
Mutation Property Assay Solvent Solvent Volume Aqueous ReferenceWT Reference Measured Value Units Solution pH Temperature Substrate(s) Product(s) Cofactor(s) Shaking Comments
WT Activity - Classical Activity of cell free extracts measured by absorbance spectrophotometry (decrease in Ξ²-Nitrostyrene absorbance measurement, 320 nm) in the presence of organic solvent Ethanol 25% 100.0 50.0 50.0 % 20 mM sodium phosphate buffer 7.3 Room temperature 50 mM Acetaldehyde, 0.5 mM Ξ²-Nitrostyrene (R)-4-Nitro-3-phenylButyraldehyde β€” 500 rpm for 60 first seconds Classical aqueous control (in %) in 5% ethanol | Categorical data/deducted temperature
WT Activity - Classical Activity of cell free extracts measured by absorbance spectrophotometry (decrease in Ξ²-Nitrostyrene absorbance measurement, 320 nm) in the presence of organic solvent Ethanol 15% 100.0 β€” 80.0 % 20 mM sodium phosphate buffer 7.3 Room temperature 50 mM Acetaldehyde, 2 mM Ξ²-Nitrostyrene (R)-4-Nitro-3-phenylButyraldehyde β€” β€” Classical aqueous control (in %) in 5% ethanol | Deducted temperature
WT Activity - Classical Activity of cell free extracts measured by absorbance spectrophotometry (decrease in Ξ²-Nitrostyrene absorbance measurement, 320 nm) in the presence of organic solvent Ethanol 25% 100.0 β€” 51.0 % 20 mM sodium phosphate buffer 7.3 Room temperature 50 mM Acetaldehyde, 2 mM Ξ²-Nitrostyrene (R)-4-Nitro-3-phenylButyraldehyde β€” β€” Classical aqueous control (in %) in 5% ethanol | Deducted temperature
WT Activity - Classical Activity of cell free extracts measured by absorbance spectrophotometry (decrease in Ξ²-Nitrostyrene absorbance measurement, 320 nm) in the presence of organic solvent Ethanol 40% 100.0 β€” 18.0 % 20 mM sodium phosphate buffer 7.3 Room temperature 50 mM Acetaldehyde, 2 mM Ξ²-Nitrostyrene (R)-4-Nitro-3-phenylButyraldehyde β€” β€” Classical aqueous control (in %) in 5% ethanol | Deducted temperature
WT Activity - Classical Activity of cell free extracts measured by absorbance spectrophotometry (decrease in Ξ²-Nitrostyrene absorbance measurement, 320 nm) in the presence of organic solvent Methanol 15% 100.0 β€” 56.0 % 20 mM sodium phosphate buffer 7.3 Room temperature 50 mM Acetaldehyde, 2 mM Ξ²-Nitrostyrene (R)-4-Nitro-3-phenylButyraldehyde β€” β€” Classical aqueous control (in %) in 5% organic solvent | Deducted temperature
WT Activity - Classical Activity of cell free extracts measured by absorbance spectrophotometry (decrease in Ξ²-Nitrostyrene absorbance measurement, 320 nm) in the presence of organic solvent Methanol 25% 100.0 β€” 37.0 % 20 mM sodium phosphate buffer 7.3 Room temperature 50 mM Acetaldehyde, 2 mM Ξ²-Nitrostyrene (R)-4-Nitro-3-phenylButyraldehyde β€” β€” Classical aqueous control (in %) in 5% organic solvent | Deducted temperature
WT Activity - Classical Activity of cell free extracts measured by absorbance spectrophotometry (decrease in Ξ²-Nitrostyrene absorbance measurement, 320 nm) in the presence of organic solvent Methanol 40% 100.0 β€” 9.0 % 20 mM sodium phosphate buffer 7.3 Room temperature 50 mM Acetaldehyde, 2 mM Ξ²-Nitrostyrene (R)-4-Nitro-3-phenylButyraldehyde β€” β€” Classical aqueous control (in %) in 5% organic solvent | Deducted temperature
WT Activity - Classical Activity of cell free extracts measured by absorbance spectrophotometry (decrease in Ξ²-Nitrostyrene absorbance measurement, 320 nm) in the presence of organic solvent 1,3-Propanediol 15% 100.0 β€” 83.0 % 20 mM sodium phosphate buffer 7.3 Room temperature 50 mM Acetaldehyde, 2 mM Ξ²-Nitrostyrene (R)-4-Nitro-3-phenylButyraldehyde β€” β€” Classical aqueous control (in %) in 5% organic solvent | Deducted temperature
WT Activity - Classical Activity of cell free extracts measured by absorbance spectrophotometry (decrease in Ξ²-Nitrostyrene absorbance measurement, 320 nm) in the presence of organic solvent 1,3-Propanediol 25% 100.0 β€” 63.0 % 20 mM sodium phosphate buffer 7.3 Room temperature 50 mM Acetaldehyde, 2 mM Ξ²-Nitrostyrene (R)-4-Nitro-3-phenylButyraldehyde β€” β€” Classical aqueous control (in %) in 5% organic solvent | Deducted temperature
WT Activity - Classical Activity of cell free extracts measured by absorbance spectrophotometry (decrease in Ξ²-Nitrostyrene absorbance measurement, 320 nm) in the presence of organic solvent 1,3-Propanediol 40% 100.0 β€” 45.0 % 20 mM sodium phosphate buffer 7.3 Room temperature 50 mM Acetaldehyde, 2 mM Ξ²-Nitrostyrene (R)-4-Nitro-3-phenylButyraldehyde β€” β€” Classical aqueous control (in %) in 5% organic solvent | Deducted temperature
WT Activity - Classical Activity of cell free extracts measured by absorbance spectrophotometry (decrease in Ξ²-Nitrostyrene absorbance measurement, 320 nm) in the presence of organic solvent Dimethyl Sulfoxide (DMSO) 15% 100.0 β€” 115.0 % 20 mM sodium phosphate buffer 7.3 Room temperature 50 mM Acetaldehyde, 2 mM Ξ²-Nitrostyrene (R)-4-Nitro-3-phenylButyraldehyde β€” β€” Classical aqueous control (in %) in 5% organic solvent | Deducted temperature
WT Activity - Classical Activity of cell free extracts measured by absorbance spectrophotometry (decrease in Ξ²-Nitrostyrene absorbance measurement, 320 nm) in the presence of organic solvent Dimethyl Sulfoxide (DMSO) 25% 100.0 β€” 119.0 % 20 mM sodium phosphate buffer 7.3 Room temperature 50 mM Acetaldehyde, 2 mM Ξ²-Nitrostyrene (R)-4-Nitro-3-phenylButyraldehyde β€” β€” Classical aqueous control (in %) in 5% organic solvent | Deducted temperature
WT Activity - Classical Activity of cell free extracts measured by absorbance spectrophotometry (decrease in Ξ²-Nitrostyrene absorbance measurement, 320 nm) in the presence of organic solvent Dimethyl Sulfoxide (DMSO) 40% 100.0 β€” 94.0 % 20 mM sodium phosphate buffer 7.3 Room temperature 50 mM Acetaldehyde, 2 mM Ξ²-Nitrostyrene (R)-4-Nitro-3-phenylButyraldehyde β€” β€” Classical aqueous control (in %) in 5% organic solvent | Deducted temperature
WT Activity - Classical Activity of cell free extracts measured by absorbance spectrophotometry (decrease in Ξ²-Nitrostyrene absorbance measurement, 320 nm) in the presence of organic solvent Isopropanol 15% 100.0 β€” 104.0 % 20 mM sodium phosphate buffer 7.3 Room temperature 50 mM Acetaldehyde, 2 mM Ξ²-Nitrostyrene (R)-4-Nitro-3-phenylButyraldehyde β€” β€” Classical aqueous control (in %) in 5% organic solvent | Deducted temperature
WT Activity - Classical Activity of cell free extracts measured by absorbance spectrophotometry (decrease in Ξ²-Nitrostyrene absorbance measurement, 320 nm) in the presence of organic solvent Isopropanol 25% 100.0 β€” 76.0 % 20 mM sodium phosphate buffer 7.3 Room temperature 50 mM Acetaldehyde, 2 mM Ξ²-Nitrostyrene (R)-4-Nitro-3-phenylButyraldehyde β€” β€” Classical aqueous control (in %) in 5% organic solvent | Deducted temperature
WT Activity - Classical Activity of cell free extracts measured by absorbance spectrophotometry (decrease in Ξ²-Nitrostyrene absorbance measurement, 320 nm) in the presence of organic solvent Isopropanol 40% 100.0 β€” 19.0 % 20 mM sodium phosphate buffer 7.3 Room temperature 50 mM Acetaldehyde, 2 mM Ξ²-Nitrostyrene (R)-4-Nitro-3-phenylButyraldehyde β€” β€” Classical aqueous control (in %) in 5% organic solvent | Deducted temperature
WT Activity - Classical Activity of cell free extracts measured by absorbance spectrophotometry (decrease in Ξ²-Nitrostyrene absorbance measurement, 320 nm) in the presence of organic solvent tert-Butanol 15% 100.0 β€” 119.0 % 20 mM sodium phosphate buffer 7.3 Room temperature 50 mM Acetaldehyde, 2 mM Ξ²-Nitrostyrene (R)-4-Nitro-3-phenylButyraldehyde β€” β€” Classical aqueous control (in %) in 5% organic solvent | Deducted temperature
WT Activity - Classical Activity of cell free extracts measured by absorbance spectrophotometry (decrease in Ξ²-Nitrostyrene absorbance measurement, 320 nm) in the presence of organic solvent tert-Butanol 25% 100.0 β€” 78.0 % 20 mM sodium phosphate buffer 7.3 Room temperature 50 mM Acetaldehyde, 2 mM Ξ²-Nitrostyrene (R)-4-Nitro-3-phenylButyraldehyde β€” β€” Classical aqueous control (in %) in 5% organic solvent | Deducted temperature
I2G Activity - Classical Activity of cell free extracts measured by absorbance spectrophotometry (decrease in Ξ²-Nitrostyrene absorbance measurement, 320 nm) in the presence of organic solvent Ethanol 25% 100.0 50.0 0.0 % 20 mM sodium phosphate buffer 7.3 Room temperature 50 mM Acetaldehyde, 0.5 mM Ξ²-Nitrostyrene (R)-4-Nitro-3-phenylButyraldehyde β€” 500 rpm for 60 first seconds Classical aqueous control (in %) in 5% ethanol | Categorical data/deducted temperature
A3G Activity - Classical Activity of cell free extracts measured by absorbance spectrophotometry (decrease in Ξ²-Nitrostyrene absorbance measurement, 320 nm) in the presence of organic solvent Ethanol 25% 100.0 50.0 50.0 % 20 mM sodium phosphate buffer 7.3 Room temperature 50 mM Acetaldehyde, 0.5 mM Ξ²-Nitrostyrene (R)-4-Nitro-3-phenylButyraldehyde β€” 500 rpm for 60 first seconds Classical aqueous control (in %) in 5% ethanol | Categorical data/deducted temperature
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1 2 3 4 … 56
Mutations in this dataset (1036)

Visualization : Activity β€” Classical

Mutation Effect

Select a condition below. Conditions with >50 mutations show a heatmap, ranked lollipop, or ranked lollipop; others show paired bars per mutation.

Structure

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