Fructose bisphosphate aldolase from Edwardsiella ictaluri
Enzyme Description
Sequence
SKIFDFVKPGVIFGDDVQKVFQVAKENKFALPAVNCVGTDSINAVMEAAAKVRAPIIVQFSNGGAAFIAGKGLKLEGQQGAILGAIAGAHHVHQMAEYYGVPVILHTDHCAKKLLPWLDGLLDAGEKHFAATGKPLFSSHMIDLSEESLEENIEICSQYLARMSKIGMTLELELGCTGGEEDGVDNSHLDNSALYTQPEDVDYAFTKLSAISPRFTIAASFGNVHGVYKPGNVQLTPVILKNSQEYVSKKHNLPHNSLNFVFHGGSGSTAAEIKEAVSYGVVKMNIDTDTQWATWEGVLKYYKKNEGYLQGQLGNPEGDDKPNKKYYDPRVWLRAAQTGMIERLEQAFKELNCIDVL
J. Hao et al. (2004)
β A thermostable variant of fructose bisphosphate aldolase constructed by directed evolution also shows increased stability in organic solvents
Protein Engineering Design and Selection
Β· doi:10.1093/protein/gzh081 β
Β· Stability - Incubation Activity + Stability - Incubation
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Database ID
UniProt: O52402 β
Sequence Annotation
Explicit - Provided (first residue from UniProt sequence absent from the mature protein)
Protein Source
Recombinant, host bacterium Escherichia coli KM3
Experimental Data (24 measurements)
24 measurements β page 1 of 2
| 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 + Stability - Incubation | Activity after incubation (1 day at room temperature) measured by absorbance spectrophotometry (glyceraldehyde-3-phosphate and hydrazone reaction product absorbance measurement, 240 nm) in the presence of organic solvent | Toluene | 20% | 79 | β | 60 | % | Incubation: 50 mM Tris-HCl buffer, 0.1M phosphate acetate, Assay: 50 mM Tris-HCl buffer, 0.1M phosphate acetate, 2 mM hydrazine dichloride hydrate | Incubation: 8, Assay: 8 | Incubation: Room temperature, Assay: 30Β°C | 4 mM Fructose 1,6-biphosphate | Dihydroxyacetone phosphate , Glyceraldehyde-3-phosphate | β | β | Water-incubated control (in %), assay in the presence of organic solvent | Clear about assay in the presence of organic solvent |
| WT | Activity + Stability - Incubation | Activity after incubation (1 day at room temperature) measured by absorbance spectrophotometry (glyceraldehyde-3-phosphate and hydrazone reaction product absorbance measurement, 240 nm) in the presence of organic solvent | n-Hexane | 20% | 79 | β | 64 | % | Incubation: 50 mM Tris-HCl buffer, 0.1M phosphate acetate, Assay: 50 mM Tris-HCl buffer, 0.1M phosphate acetate, 2 mM hydrazine dichloride hydrate | Incubation: 8, Assay: 8 | Incubation: Room temperature, Assay: 30Β°C | 4 mM Fructose 1,6-biphosphate | Dihydroxyacetone phosphate , Glyceraldehyde-3-phosphate | β | β | Water-incubated control (in %), assay in the presence of organic solvent | Clear about assay in the presence of organic solvent |
| WT | Activity + Stability - Incubation | Activity after incubation (1 day at room temperature) measured by absorbance spectrophotometry (glyceraldehyde-3-phosphate and hydrazone reaction product absorbance measurement, 240 nm) in the presence of organic solvent | Dimethylformamide (DMF) | 20% | 79 | β | 4 | % | Incubation: 50 mM Tris-HCl buffer, 0.1M phosphate acetate, Assay: 50 mM Tris-HCl buffer, 0.1M phosphate acetate, 2 mM hydrazine dichloride hydrate | Incubation: 8, Assay: 8 | Incubation: Room temperature, Assay: 30Β°C | 4 mM Fructose 1,6-biphosphate | Dihydroxyacetone phosphate , Glyceraldehyde-3-phosphate | β | β | Water-incubated control (in %), assay in the presence of organic solvent | Clear about assay in the presence of organic solvent |
| WT | Activity + Stability - Incubation | Activity after incubation (1 day at room temperature) measured by absorbance spectrophotometry (glyceraldehyde-3-phosphate and hydrazone reaction product absorbance measurement, 240 nm) in the presence of organic solvent | Acetonitrile | 20% | 79 | β | 18 | % | Incubation: 50 mM Tris-HCl buffer, 0.1M phosphate acetate, Assay: 50 mM Tris-HCl buffer, 0.1M phosphate acetate, 2 mM hydrazine dichloride hydrate | Incubation: 8, Assay: 8 | Incubation: Room temperature, Assay: 30Β°C | 4 mM Fructose 1,6-biphosphate | Dihydroxyacetone phosphate , Glyceraldehyde-3-phosphate | β | β | Water-incubated control (in %), assay in the presence of organic solvent | Clear about assay in the presence of organic solvent |
| WT | Activity + Stability - Incubation | Activity after incubation (1 day at room temperature) measured by absorbance spectrophotometry (glyceraldehyde-3-phosphate and hydrazone reaction product absorbance measurement, 240 nm) in the presence of organic solvent | Chloroform | 20% | 79 | β | 21 | % | Incubation: 50 mM Tris-HCl buffer, 0.1M phosphate acetate, Assay: 50 mM Tris-HCl buffer, 0.1M phosphate acetate, 2 mM hydrazine dichloride hydrate | Incubation: 8, Assay: 8 | Incubation: Room temperature, Assay: 30Β°C | 4 mM Fructose 1,6-biphosphate | Dihydroxyacetone phosphate , Glyceraldehyde-3-phosphate | β | β | Water-incubated control (in %), assay in the presence of organic solvent | Clear about assay in the presence of organic solvent |
| WT | Activity + Stability - Incubation | Activity after incubation (1 day at room temperature) measured by absorbance spectrophotometry (glyceraldehyde-3-phosphate and hydrazone reaction product absorbance measurement, 240 nm) in the presence of organic solvent | Methanol | 20% | 79 | β | 58 | % | Incubation: 50 mM Tris-HCl buffer, 0.1M phosphate acetate, Assay: 50 mM Tris-HCl buffer, 0.1M phosphate acetate, 2 mM hydrazine dichloride hydrate | Incubation: 8, Assay: 8 | Incubation: Room temperature, Assay: 30Β°C | 4 mM Fructose 1,6-biphosphate | Dihydroxyacetone phosphate , Glyceraldehyde-3-phosphate | β | β | Water-incubated control (in %), assay in the presence of organic solvent | Clear about assay in the presence of organic solvent |
| WT | Stability - Incubation | Activity after incubation (1 day at room temperature) measured by absorbance spectrophotometry (glyceraldehyde-3-phosphate and hydrazone reaction product absorbance measurement, 240 nm) in aqueous phase | Methanol | 20% | 79 | β | 50 | % | Incubation: 50 mM Tris-HCl buffer, 0.1M phosphate acetate, Assay: 50 mM Tris-HCl buffer, 0.1M phosphate acetate, 2 mM hydrazine dichloride hydrate | Incubation: 8, Assay: 8 | Incubation: Room temperature, Assay: 30Β°C | 4 mM Fructose 1,6-biphosphate | Dihydroxyacetone phosphate , Glyceraldehyde-3-phosphate | β | β | Water-incubated control (in %), assay in aqueous phase | Clear about assay in aqueous phase |
| WT | Stability - Incubation | Activity after incubation (1 day at room temperature) measured by absorbance spectrophotometry (glyceraldehyde-3-phosphate and hydrazone reaction product absorbance measurement, 240 nm) in aqueous phase | Chloroform | 20% | 79 | β | 50 | % | Incubation: 50 mM Tris-HCl buffer, 0.1M phosphate acetate, Assay: 50 mM Tris-HCl buffer, 0.1M phosphate acetate, 2 mM hydrazine dichloride hydrate | Incubation: 8, Assay: 8 | Incubation: Room temperature, Assay: 30Β°C | 4 mM Fructose 1,6-biphosphate | Dihydroxyacetone phosphate , Glyceraldehyde-3-phosphate | β | β | Water-incubated control (in %), assay in aqueous phase | Clear about assay in aqueous phase |
| WT | Stability - Incubation | Activity after incubation (1 day at room temperature) measured by absorbance spectrophotometry (glyceraldehyde-3-phosphate and hydrazone reaction product absorbance measurement, 240 nm) in aqueous phase | Acetonitrile | 20% | 79 | β | 7 | % | Incubation: 50 mM Tris-HCl buffer, 0.1M phosphate acetate, Assay: 50 mM Tris-HCl buffer, 0.1M phosphate acetate, 2 mM hydrazine dichloride hydrate | Incubation: 8, Assay: 8 | Incubation: Room temperature, Assay: 30Β°C | 4 mM Fructose 1,6-biphosphate | Dihydroxyacetone phosphate , Glyceraldehyde-3-phosphate | β | β | Water-incubated control (in %), assay in aqueous phase | Clear about assay in aqueous phase |
| WT | Stability - Incubation | Activity after incubation (1 day at room temperature) measured by absorbance spectrophotometry (glyceraldehyde-3-phosphate and hydrazone reaction product absorbance measurement, 240 nm) in aqueous phase | Dimethylformamide (DMF) | 20% | 79 | β | 20 | % | Incubation: 50 mM Tris-HCl buffer, 0.1M phosphate acetate, Assay: 50 mM Tris-HCl buffer, 0.1M phosphate acetate, 2 mM hydrazine dichloride hydrate | Incubation: 8, Assay: 8 | Incubation: Room temperature, Assay: 30Β°C | 4 mM Fructose 1,6-biphosphate | Dihydroxyacetone phosphate , Glyceraldehyde-3-phosphate | β | β | Water-incubated control (in %), assay in aqueous phase | Clear about assay in aqueous phase |
| WT | Stability - Incubation | Activity after incubation (1 day at room temperature) measured by absorbance spectrophotometry (glyceraldehyde-3-phosphate and hydrazone reaction product absorbance measurement, 240 nm) in aqueous phase | n-Hexane | 20% | 79 | β | 66 | % | Incubation: 50 mM Tris-HCl buffer, 0.1M phosphate acetate, Assay: 50 mM Tris-HCl buffer, 0.1M phosphate acetate, 2 mM hydrazine dichloride hydrate | Incubation: 8, Assay: 8 | Incubation: Room temperature, Assay: 30Β°C | 4 mM Fructose 1,6-biphosphate | Dihydroxyacetone phosphate , Glyceraldehyde-3-phosphate | β | β | Water-incubated control (in %), assay in aqueous phase | Clear about assay in aqueous phase |
| WT | Stability - Incubation | Activity after incubation (1 day at room temperature) measured by absorbance spectrophotometry (glyceraldehyde-3-phosphate and hydrazone reaction product absorbance measurement, 240 nm) in aqueous phase | Toluene | 20% | 79 | β | 57 | % | Incubation: 50 mM Tris-HCl buffer, 0.1M phosphate acetate, Assay: 50 mM Tris-HCl buffer, 0.1M phosphate acetate, 2 mM hydrazine dichloride hydrate | Incubation: 8, Assay: 8 | Incubation: Room temperature, Assay: 30Β°C | 4 mM Fructose 1,6-biphosphate | Dihydroxyacetone phosphate , Glyceraldehyde-3-phosphate | β | β | Water-incubated control (in %), assay in aqueous phase | Clear about assay in aqueous phase |
| V45A/K71I/Q79L/A111T/A124V/A210V/Q346R | Activity + Stability - Incubation | Activity after incubation (1 day at room temperature) measured by absorbance spectrophotometry (glyceraldehyde-3-phosphate and hydrazone reaction product absorbance measurement, 240 nm) in the presence of organic solvent | Toluene | 20% | 96 | 60 | 86 | % | Incubation: 50 mM Tris-HCl buffer, 0.1M phosphate acetate, Assay: 50 mM Tris-HCl buffer, 0.1M phosphate acetate, 2 mM hydrazine dichloride hydrate | Incubation: 8, Assay: 8 | Incubation: Room temperature, Assay: 30Β°C | 4 mM Fructose 1,6-biphosphate | Dihydroxyacetone phosphate , Glyceraldehyde-3-phosphate | β | β | Water-incubated control (in %), assay in the presence of organic solvent | Clear about assay in the presence of organic solvent |
| V45A/K71I/Q79L/A111T/A124V/A210V/Q346R | Activity + Stability - Incubation | Activity after incubation (1 day at room temperature) measured by absorbance spectrophotometry (glyceraldehyde-3-phosphate and hydrazone reaction product absorbance measurement, 240 nm) in the presence of organic solvent | Methanol | 20% | 96 | 58 | 102 | % | Incubation: 50 mM Tris-HCl buffer, 0.1M phosphate acetate, Assay: 50 mM Tris-HCl buffer, 0.1M phosphate acetate, 2 mM hydrazine dichloride hydrate | Incubation: 8, Assay: 8 | Incubation: Room temperature, Assay: 30Β°C | 4 mM Fructose 1,6-biphosphate | Dihydroxyacetone phosphate , Glyceraldehyde-3-phosphate | β | β | Water-incubated control (in %), assay in the presence of organic solvent | Clear about assay in the presence of organic solvent |
| V45A/K71I/Q79L/A111T/A124V/A210V/Q346R | Activity + Stability - Incubation | Activity after incubation (1 day at room temperature) measured by absorbance spectrophotometry (glyceraldehyde-3-phosphate and hydrazone reaction product absorbance measurement, 240 nm) in the presence of organic solvent | Chloroform | 20% | 96 | 21 | 84 | % | Incubation: 50 mM Tris-HCl buffer, 0.1M phosphate acetate, Assay: 50 mM Tris-HCl buffer, 0.1M phosphate acetate, 2 mM hydrazine dichloride hydrate | Incubation: 8, Assay: 8 | Incubation: Room temperature, Assay: 30Β°C | 4 mM Fructose 1,6-biphosphate | Dihydroxyacetone phosphate , Glyceraldehyde-3-phosphate | β | β | Water-incubated control (in %), assay in the presence of organic solvent | Clear about assay in the presence of organic solvent |
| V45A/K71I/Q79L/A111T/A124V/A210V/Q346R | Activity + Stability - Incubation | Activity after incubation (1 day at room temperature) measured by absorbance spectrophotometry (glyceraldehyde-3-phosphate and hydrazone reaction product absorbance measurement, 240 nm) in the presence of organic solvent | Acetonitrile | 20% | 96 | 18 | 53 | % | Incubation: 50 mM Tris-HCl buffer, 0.1M phosphate acetate, Assay: 50 mM Tris-HCl buffer, 0.1M phosphate acetate, 2 mM hydrazine dichloride hydrate | Incubation: 8, Assay: 8 | Incubation: Room temperature, Assay: 30Β°C | 4 mM Fructose 1,6-biphosphate | Dihydroxyacetone phosphate , Glyceraldehyde-3-phosphate | β | β | Water-incubated control (in %), assay in the presence of organic solvent | Clear about assay in the presence of organic solvent |
| V45A/K71I/Q79L/A111T/A124V/A210V/Q346R | Activity + Stability - Incubation | Activity after incubation (1 day at room temperature) measured by absorbance spectrophotometry (glyceraldehyde-3-phosphate and hydrazone reaction product absorbance measurement, 240 nm) in the presence of organic solvent | Dimethylformamide (DMF) | 20% | 96 | 4 | 8 | % | Incubation: 50 mM Tris-HCl buffer, 0.1M phosphate acetate, Assay: 50 mM Tris-HCl buffer, 0.1M phosphate acetate, 2 mM hydrazine dichloride hydrate | Incubation: 8, Assay: 8 | Incubation: Room temperature, Assay: 30Β°C | 4 mM Fructose 1,6-biphosphate | Dihydroxyacetone phosphate , Glyceraldehyde-3-phosphate | β | β | Water-incubated control (in %), assay in the presence of organic solvent | Clear about assay in the presence of organic solvent |
| V45A/K71I/Q79L/A111T/A124V/A210V/Q346R | Activity + Stability - Incubation | Activity after incubation (1 day at room temperature) measured by absorbance spectrophotometry (glyceraldehyde-3-phosphate and hydrazone reaction product absorbance measurement, 240 nm) in the presence of organic solvent | n-Hexane | 20% | 96 | 64 | 87 | % | Incubation: 50 mM Tris-HCl buffer, 0.1M phosphate acetate, Assay: 50 mM Tris-HCl buffer, 0.1M phosphate acetate, 2 mM hydrazine dichloride hydrate | Incubation: 8, Assay: 8 | Incubation: Room temperature, Assay: 30Β°C | 4 mM Fructose 1,6-biphosphate | Dihydroxyacetone phosphate , Glyceraldehyde-3-phosphate | β | β | Water-incubated control (in %), assay in the presence of organic solvent | Clear about assay in the presence of organic solvent |
| V45A/K71I/Q79L/A111T/A124V/A210V/Q346R | Stability - Incubation | Activity after incubation (1 day at room temperature) measured by absorbance spectrophotometry (glyceraldehyde-3-phosphate and hydrazone reaction product absorbance measurement, 240 nm) in aqueous phase | Toluene | 20% | 96 | 57 | 102 | % | Incubation: 50 mM Tris-HCl buffer, 0.1M phosphate acetate, Assay: 50 mM Tris-HCl buffer, 0.1M phosphate acetate, 2 mM hydrazine dichloride hydrate | Incubation: 8, Assay: 8 | Incubation: Room temperature, Assay: 30Β°C | 4 mM Fructose 1,6-biphosphate | Dihydroxyacetone phosphate , Glyceraldehyde-3-phosphate | β | β | Water-incubated control (in %), assay in aqueous phase | Clear about assay in aqueous phase |
| V45A/K71I/Q79L/A111T/A124V/A210V/Q346R | Stability - Incubation | Activity after incubation (1 day at room temperature) measured by absorbance spectrophotometry (glyceraldehyde-3-phosphate and hydrazone reaction product absorbance measurement, 240 nm) in aqueous phase | n-Hexane | 20% | 96 | 66 | 100 | % | Incubation: 50 mM Tris-HCl buffer, 0.1M phosphate acetate, Assay: 50 mM Tris-HCl buffer, 0.1M phosphate acetate, 2 mM hydrazine dichloride hydrate | Incubation: 8, Assay: 8 | Incubation: Room temperature, Assay: 30Β°C | 4 mM Fructose 1,6-biphosphate | Dihydroxyacetone phosphate , Glyceraldehyde-3-phosphate | β | β | Water-incubated control (in %), assay in aqueous phase | Clear about assay in aqueous phase |
Mutations in this dataset (2)
WT
V45A/K71I/Q79L/A111T/A124V/A210V/Q346R
Visualization : Stability β Incubation
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.
Visualization : Activity + Stability β Incubation
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.