Carboxylesterase from Geobacillus kaustophilus HTA426
Enzyme Description
Extremophile
Yes
thermophilic organism
EC Number
Sequence
MKIVPPKPFFFEAGERAVLLLHGFTGNSADVRMLGRFLESKGYTCHAPIYKGHGVPPEELVHTGPDDWWQDVMNGYQFLKNKGYEKIAVAGLSLGGVFSLKLGYTVPTQGIVTMCAPMYIKSEETMYEGVLEYAREYKKREGKSEEQIEQEMERFKQTPMKTLKALQELIADVRAHLDLVYAPTFVVQARHDEMINPDSANIIYNEIESPVKQIKWYEQSGHVITLDQEKDQLHEDIYAFLESLDW
Silvia Montoro-GarciΜa et al. (2009)
β Characterization of a Novel Thermostable Carboxylesterase from Geobacillus kaustophilus HTA426 Shows the Existence of a New Carboxylesterase Family
Journal of Bacteriology
Β· doi:10.1128/JB.01060-08 β
Β· Activity - Classical Stability - Incubation
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Database ID
UniProt: Q2V6P5 β
Sequence Annotation
Explicit - Provided (first residue removed from UniProt sequence from mature protein, 1 mutation)
Protein Source
Recombinant, host bacterium Escherichia coli Rosetta (DE3)
Experimental Data (36 measurements)
36 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 |
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| T108I | Activity - Classical | Activity measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in the presence of organic solvent | Methanol | 90% | 100 | β | 73 | % | 50 mM sodium phosphate buffer, 0.36% Triton X-100, 0.1% gum arabic | 8 | 30Β°C | 0.8 mM p-Nitrophenyl caprylate | Caprylic Acid , p-Nitrophenol | β | β | Classical aqueous control (in %) |
| T108I | Activity - Classical | Activity measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in the presence of organic solvent | Methanol | 40% | 100 | β | 103 | % | 50 mM sodium phosphate buffer, 0.36% Triton X-100, 0.1% gum arabic | 8 | 30Β°C | 0.8 mM p-Nitrophenyl caprylate | Caprylic Acid , p-Nitrophenol | β | β | Classical aqueous control (in %) |
| T108I | Activity - Classical | Activity measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in the presence of organic solvent | tert-Butyl Methyl Ether (MTBE) | 90% | 100 | β | 33 | % | 50 mM sodium phosphate buffer, 0.36% Triton X-100, 0.1% gum arabic | 8 | 30Β°C | 0.8 mM p-Nitrophenyl caprylate | Caprylic Acid , p-Nitrophenol | β | β | Classical aqueous control (in %) |
| T108I | Activity - Classical | Activity measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in the presence of organic solvent | tert-Butyl Methyl Ether (MTBE) | 40% | 100 | β | 44 | % | 50 mM sodium phosphate buffer, 0.36% Triton X-100, 0.1% gum arabic | 8 | 30Β°C | 0.8 mM p-Nitrophenyl caprylate | Caprylic Acid , p-Nitrophenol | β | β | Classical aqueous control (in %) |
| T108I | Activity - Classical | Activity measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in the presence of organic solvent | Dimethyl Sulfoxide (DMSO) | 90% | 100 | β | 62 | % | 50 mM sodium phosphate buffer, 0.36% Triton X-100, 0.1% gum arabic | 8 | 30Β°C | 0.8 mM p-Nitrophenyl caprylate | Caprylic Acid , p-Nitrophenol | β | β | Classical aqueous control (in %) |
| T108I | Activity - Classical | Activity measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in the presence of organic solvent | Dimethyl Sulfoxide (DMSO) | 40% | 100 | β | 97 | % | 50 mM sodium phosphate buffer, 0.36% Triton X-100, 0.1% gum arabic | 8 | 30Β°C | 0.8 mM p-Nitrophenyl caprylate | Caprylic Acid , p-Nitrophenol | β | β | Classical aqueous control (in %) |
| T108I | Activity - Classical | Activity measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in the presence of organic solvent | Isopropanol | 90% | 100 | β | 87 | % | 50 mM sodium phosphate buffer, 0.36% Triton X-100, 0.1% gum arabic | 8 | 30Β°C | 0.8 mM p-Nitrophenyl caprylate | Caprylic Acid , p-Nitrophenol | β | β | Classical aqueous control (in %) |
| T108I | Activity - Classical | Activity measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in the presence of organic solvent | Acetone | 90% | 100 | β | 50 | % | 50 mM sodium phosphate buffer, 0.36% Triton X-100, 0.1% gum arabic | 8 | 30Β°C | 0.8 mM p-Nitrophenyl caprylate | Caprylic Acid , p-Nitrophenol | β | β | Classical aqueous control (in %) |
| T108I | Activity - Classical | Activity measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in the presence of organic solvent | Acetone | 40% | 100 | β | 74 | % | 50 mM sodium phosphate buffer, 0.36% Triton X-100, 0.1% gum arabic | 8 | 30Β°C | 0.8 mM p-Nitrophenyl caprylate | Caprylic Acid , p-Nitrophenol | β | β | Classical aqueous control (in %) |
| T108I | Activity - Classical | Activity measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in the presence of organic solvent | Ethanol | 90% | 100 | β | 94 | % | 50 mM sodium phosphate buffer, 0.36% Triton X-100, 0.1% gum arabic | 8 | 30Β°C | 0.8 mM p-Nitrophenyl caprylate | Caprylic Acid , p-Nitrophenol | β | β | Classical aqueous control (in %) |
| T108I | Activity - Classical | Activity measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in the presence of organic solvent | Ethanol | 40% | 100 | β | 86 | % | 50 mM sodium phosphate buffer, 0.36% Triton X-100, 0.1% gum arabic | 8 | 30Β°C | 0.8 mM p-Nitrophenyl caprylate | Caprylic Acid , p-Nitrophenol | β | β | Classical aqueous control (in %) |
| T108I | Activity - Classical | Activity measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in the presence of organic solvent | Isopropanol | 40% | 100 | β | 88 | % | 50 mM sodium phosphate buffer, 0.36% Triton X-100, 0.1% gum arabic | 8 | 30Β°C | 0.8 mM p-Nitrophenyl caprylate | Caprylic Acid , p-Nitrophenol | β | β | Classical aqueous control (in %) |
| T108I | Stability - Incubation | Activity after incubation (1 hour at 60Β°C) measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in aqueous phase | Methanol | 40% | 100 | β | 3 | % | Incubation: 50 mM sodium phosphate buffer, Assay: 50 mM sodium phosphate buffer, 0.36% Triton X-100, 0.1% gum arabic | Incubation: 8, Assay: 8 | Incubation: 60Β°C, Assay: 30Β°C | 0.8 mM p-Nitrophenyl caprylate | Caprylic Acid , p-Nitrophenol | β | β | Water-incubated control (in %), activity measured in the absence of organic solvent |
| T108I | Stability - Incubation | Activity after incubation (1 hour at 60Β°C) measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in aqueous phase | Methanol | 90% | 100 | β | 0 | % | Incubation: 50 mM sodium phosphate buffer, Assay: 50 mM sodium phosphate buffer, 0.36% Triton X-100, 0.1% gum arabic | Incubation: 8, Assay: 8 | Incubation: 60Β°C, Assay: 30Β°C | 0.8 mM p-Nitrophenyl caprylate | Caprylic Acid , p-Nitrophenol | β | β | Water-incubated control (in %), activity measured in the absence of organic solvent |
| T108I | Stability - Incubation | Activity after incubation (1 hour at 60Β°C) measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in aqueous phase | Ethanol | 40% | 100 | β | 8 | % | Incubation: 50 mM sodium phosphate buffer, Assay: 50 mM sodium phosphate buffer, 0.36% Triton X-100, 0.1% gum arabic | Incubation: 8, Assay: 8 | Incubation: 60Β°C, Assay: 30Β°C | 0.8 mM p-Nitrophenyl caprylate | Caprylic Acid , p-Nitrophenol | β | β | Water-incubated control (in %), activity measured in the absence of organic solvent |
| T108I | Stability - Incubation | Activity after incubation (1 hour at 60Β°C) measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in aqueous phase | Ethanol | 90% | 100 | β | 0 | % | Incubation: 50 mM sodium phosphate buffer, Assay: 50 mM sodium phosphate buffer, 0.36% Triton X-100, 0.1% gum arabic | Incubation: 8, Assay: 8 | Incubation: 60Β°C, Assay: 30Β°C | 0.8 mM p-Nitrophenyl caprylate | Caprylic Acid , p-Nitrophenol | β | β | Water-incubated control (in %), activity measured in the absence of organic solvent |
| T108I | Stability - Incubation | Activity after incubation (1 hour at 60Β°C) measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in aqueous phase | Acetone | 40% | 100 | β | 2 | % | Incubation: 50 mM sodium phosphate buffer, Assay: 50 mM sodium phosphate buffer, 0.36% Triton X-100, 0.1% gum arabic | Incubation: 8, Assay: 8 | Incubation: 60Β°C, Assay: 30Β°C | 0.8 mM p-Nitrophenyl caprylate | Caprylic Acid , p-Nitrophenol | β | β | Water-incubated control (in %), activity measured in the absence of organic solvent |
| T108I | Stability - Incubation | Activity after incubation (1 hour at 60Β°C) measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in aqueous phase | Acetone | 90% | 100 | β | 2 | % | Incubation: 50 mM sodium phosphate buffer, Assay: 50 mM sodium phosphate buffer, 0.36% Triton X-100, 0.1% gum arabic | Incubation: 8, Assay: 8 | Incubation: 60Β°C, Assay: 30Β°C | 0.8 mM p-Nitrophenyl caprylate | Caprylic Acid , p-Nitrophenol | β | β | Water-incubated control (in %), activity measured in the absence of organic solvent |
| T108I | Stability - Incubation | Activity after incubation (1 hour at 60Β°C) measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in aqueous phase | Isopropanol | 40% | 100 | β | 16 | % | Incubation: 50 mM sodium phosphate buffer, Assay: 50 mM sodium phosphate buffer, 0.36% Triton X-100, 0.1% gum arabic | Incubation: 8, Assay: 8 | Incubation: 60Β°C, Assay: 30Β°C | 0.8 mM p-Nitrophenyl caprylate | Caprylic Acid , p-Nitrophenol | β | β | Water-incubated control (in %), activity measured in the absence of organic solvent |
| T108I | Stability - Incubation | Activity after incubation (1 hour at 60Β°C) measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in aqueous phase | Isopropanol | 90% | 100 | β | 0 | % | Incubation: 50 mM sodium phosphate buffer, Assay: 50 mM sodium phosphate buffer, 0.36% Triton X-100, 0.1% gum arabic | Incubation: 8, Assay: 8 | Incubation: 60Β°C, Assay: 30Β°C | 0.8 mM p-Nitrophenyl caprylate | Caprylic Acid , p-Nitrophenol | β | β | Water-incubated control (in %), activity measured in the absence of organic solvent |
Mutations in this dataset (1)
T108I
Visualization : Activity β Classical
One bar per measurement. Colour = solvent, shade = solvent volume.
Visualization : Stability β Incubation
One bar per measurement. Colour = solvent, shade = solvent volume.