Lipase BTL2 from Bacillus thermocatenulatus with a pentapeptide
Enzyme Description
Sequence
AASPRANDAPIVLLHGFTGWGREEMLGFKYWGGVRGDIEQWLNDNGYRTYTLAVGPLSSNWDRACEAYAQLVGGTVDYGAAHAAKHGHARFGRTYPGLLPELKRGGRVHIIAHSQGGQTARMLVSLLENGSQEEREYAKAHNVSLSPLFEGGHHFVLSVTTIATPHDGTTLVNMVDFTDRFFDLQKAVLKAAAVASNVPYTSQVYDFKLDQWGLRRQPGESFDHYFERLKRSPVWTSTDTARYDLSIPGAEKLNQWVQASPNTYYLSFSTERTHRGALTGNYYPELGMNAFSAVVCAPFLGSYRNEALGIDDRWLENDGIVNTVSMNGPKRGSSDRIVPYDGTLKKGVWNDMGTCNVDHLEVIGVDPNPSFDIRAFYLRLAEQLASLRP
Mostafa Hosseini et al. (2012)
β In Silico and Experimental Characterization of Chimeric Bacillus thermocatenulatus Lipase with the Complete Conserved Pentapeptide of Candida rugosa Lipase
Applied Biochemistry and Biotechnology
Β· doi:10.1007/s12010-012-0014-0 β
Β· Stability - Incubation
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Database ID
UniProt: Q59260 β
Sequence Annotation
Explicit - Provided GenBank Accession Number (28 residues from UniProt sequence absent from the mature protein)
Protein Source
Recombinant, host bacterium Escherichia coli TOP10
Experimental Data (10 measurements)
10 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 | Stability - Incubation | Activity after incubation (1 hour at room temperature), measured by pH-stat (titration of the released fatty acids with NaOH) in aqueous phase | Acetone | 30% | 100 | β | 97 | % | Incubation: 50 mM Tris-HCl, Assay: 50 mM Tris-HCl, 20 mg/ml arabic gum | Incubation: 8, Assay: 8 | Incubation: Room temperature, Assay: 55Β°C | 2 mg/mL Tricaprylin | Caprylic Acid , glycerol | β | β | Non-incubated control (in %), activity measured in aqueous phase | Uncertainty about whether activity was measured with or without organic solvent |
| WT | Stability - Incubation | Activity after incubation (1 hour at room temperature), measured by pH-stat (titration of the released fatty acids with NaOH) in aqueous phase | Methanol | 30% | 100 | β | 96 | % | Incubation: 50 mM Tris-HCl, Assay: 50 mM Tris-HCl, 20 mg/ml arabic gum | Incubation: 8, Assay: 8 | Incubation: Room temperature, Assay: 55Β°C | 2 mg/mL Tricaprylin | Caprylic Acid , glycerol | β | β | Non-incubated control (in %), activity measured in aqueous phase | Uncertainty about whether activity was measured with or without organic solvent |
| WT | Stability - Incubation | Activity after incubation (1 hour at room temperature), measured by pH-stat (titration of the released fatty acids with NaOH) in aqueous phase | n-Heptane | 30% | 100 | β | 106 | % | Incubation: 50 mM Tris-HCl, Assay: 50 mM Tris-HCl, 20 mg/ml arabic gum | Incubation: 8, Assay: 8 | Incubation: Room temperature, Assay: 55Β°C | 2 mg/mL Tricaprylin | Caprylic Acid , glycerol | β | β | Non-incubated control (in %), activity measured in aqueous phase | Uncertainty about whether activity was measured with or without organic solvent |
| WT | Stability - Incubation | Activity after incubation (1 hour at room temperature), measured by pH-stat (titration of the released fatty acids with NaOH) in aqueous phase | n-Hexane | 30% | 100 | β | 118 | % | Incubation: 50 mM Tris-HCl, Assay: 50 mM Tris-HCl, 20 mg/ml arabic gum | Incubation: 8, Assay: 8 | Incubation: Room temperature, Assay: 55Β°C | 2 mg/mL Tricaprylin | Caprylic Acid , glycerol | β | β | Non-incubated control (in %), activity measured in aqueous phase | Uncertainty about whether activity was measured with or without organic solvent |
| WT | Stability - Incubation | Activity after incubation (1 hour at room temperature), measured by pH-stat (titration of the released fatty acids with NaOH) in aqueous phase | Chloroform | 30% | 100 | β | 129 | % | Incubation: 50 mM Tris-HCl, Assay: 50 mM Tris-HCl, 20 mg/ml arabic gum | Incubation: 8, Assay: 8 | Incubation: Room temperature, Assay: 55Β°C | 2 mg/mL Tricaprylin | Caprylic Acid , glycerol | β | β | Non-incubated control (in %), activity measured in aqueous phase | Uncertainty about whether activity was measured with or without organic solvent |
| A112G/H113E/Q115A | Stability - Incubation | Activity after incubation (1 hour at room temperature), measured by pH-stat (titration of the released fatty acids with NaOH) in aqueous phase | Acetone | 30% | 100 | 97 | 141 | % | Incubation: 50 mM Tris-HCl, Assay: 50 mM Tris-HCl, 20 mg/ml arabic gum | Incubation: 8, Assay: 8 | Incubation: Room temperature, Assay: 55Β°C | 2 mg/mL Tricaprylin | Caprylic Acid , glycerol | β | β | Non-incubated control (in %), activity measured in aqueous phase | Uncertainty about whether activity was measured with or without organic solvent |
| A112G/H113E/Q115A | Stability - Incubation | Activity after incubation (1 hour at room temperature), measured by pH-stat (titration of the released fatty acids with NaOH) in aqueous phase | Methanol | 30% | 100 | 96 | 138 | % | Incubation: 50 mM Tris-HCl, Assay: 50 mM Tris-HCl, 20 mg/ml arabic gum | Incubation: 8, Assay: 8 | Incubation: Room temperature, Assay: 55Β°C | 2 mg/mL Tricaprylin | Caprylic Acid , glycerol | β | β | Non-incubated control (in %), activity measured in aqueous phase | Uncertainty about whether activity was measured with or without organic solvent |
| A112G/H113E/Q115A | Stability - Incubation | Activity after incubation (1 hour at room temperature), measured by pH-stat (titration of the released fatty acids with NaOH) in aqueous phase | n-Heptane | 30% | 100 | 106 | 161 | % | Incubation: 50 mM Tris-HCl, Assay: 50 mM Tris-HCl, 20 mg/ml arabic gum | Incubation: 8, Assay: 8 | Incubation: Room temperature, Assay: 55Β°C | 2 mg/mL Tricaprylin | Caprylic Acid , glycerol | β | β | Non-incubated control (in %), activity measured in aqueous phase | Uncertainty about whether activity was measured with or without organic solvent |
| A112G/H113E/Q115A | Stability - Incubation | Activity after incubation (1 hour at room temperature), measured by pH-stat (titration of the released fatty acids with NaOH) in aqueous phase | n-Hexane | 30% | 100 | 118 | 157 | % | Incubation: 50 mM Tris-HCl, Assay: 50 mM Tris-HCl, 20 mg/ml arabic gum | Incubation: 8, Assay: 8 | Incubation: Room temperature, Assay: 55Β°C | 2 mg/mL Tricaprylin | Caprylic Acid , glycerol | β | β | Non-incubated control (in %), activity measured in aqueous phase | Uncertainty about whether activity was measured with or without organic solvent |
| A112G/H113E/Q115A | Stability - Incubation | Activity after incubation (1 hour at room temperature), measured by pH-stat (titration of the released fatty acids with NaOH) in aqueous phase | Chloroform | 30% | 100 | 129 | 160 | % | Incubation: 50 mM Tris-HCl, Assay: 50 mM Tris-HCl, 20 mg/ml arabic gum | Incubation: 8, Assay: 8 | Incubation: Room temperature, Assay: 55Β°C | 2 mg/mL Tricaprylin | Caprylic Acid , glycerol | β | β | Non-incubated control (in %), activity measured in aqueous phase | Uncertainty about whether activity was measured with or without organic solvent |
Mutations in this dataset (2)
WT
A112G/H113E/Q115A
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.