Glycosylated lipase BTL2 from Bacillus thermocatenulatus
Enzyme Description
Species
Extremophile
No
but "thermostable" enzyme
EC Number
Sequence
MASPRANDAPIVLLHGFTGWGREEMLGFKYWGGVRGDIEQWLNDNGYRTYTLAVGPLSSNWDRACEAYAQLVGGTVDYGAAHAAKHGHARFGRTYPGLLPELKRGGRVHIIAHSQGGQTARMLVSLLENGSQEEREYAKAHNVSLSPLFEGGHHFVLSVTTIATPHDGTTLVNMVDFTDRFFDLQKAVLKAAAVASNVPYTSQVYDFKLDQWGLRRQPGESFDHYFERLKRSPVWTSTDTARYDLSIPGAEKLNQWVQASPNTYYLSFSTERTHRGALTGNYYPELGMNAFSAVVCAPFLGSYRNEALGIDDRWLENDGIVNTVSMNGPKRGSSDRIVPYDGTLKKGVWNDMGTYNVDHLEVIGVDPNPSFDIRAFYLRLAEQLASLRP
Shota Kajiwara et al. (2019)
β N-linked glycosylation of thermostable lipase from Bacillus thermocatenulatus to improve organic solvent stability
Enzyme and Microbial Technology
Β· doi:10.1016/j.enzmictec.2019.109416 β
Β· Stability - Incubation
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Database ID
PDB: 2W22 β
Sequence Annotation
Explicit - Provided PDB Accession Number
Protein Source
Recombinant, host yeast Saccharomyces cerevisiae YPH499
Experimental Data (7 measurements)
7 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 (20 hours at 45Β°C), measured by colorimetric assay with Ellman's reagent (5-thio-2-nitrobenzoate anion absorbance measurement, 412 nm) in aqueous phase | Dimethyl Sulfoxide (DMSO) | 25% | 100.0 | β | 56.1 | % | Incubation: 100 mM Tris-HCl buffer, Assay: 100 mM Tris-HCl buffer | Incubation: 8, Assay: 8 | Incubation: 45Β°C, Assay: 30Β°C | ? mM 2,3-Dimercapto-1-propanol tributyrate | butyrate , Free thiols | β | Incubation: 160 rpm | Non-incubated control (in %), assay in aqueous phase | Uncertainty about whether assay really was in aqueous phase |
| A277N | Stability - Incubation | Activity after incubation (20 hours at 45Β°C), measured by colorimetric assay with Ellman's reagent (5-thio-2-nitrobenzoate anion absorbance measurement, 412 nm) in aqueous phase | Dimethyl Sulfoxide (DMSO) | 25% | 100.0 | 56.1 | 74.3 | % | Incubation: 100 mM Tris-HCl buffer, Assay: 100 mM Tris-HCl buffer | Incubation: 8, Assay: 8 | Incubation: 45Β°C, Assay: 30Β°C | ? mM 2,3-Dimercapto-1-propanol tributyrate | butyrate , Free thiols | β | Incubation: 160 rpm | Non-incubated control (in %), assay in aqueous phase | Uncertainty about whether assay really was in aqueous phase |
| A290N | Stability - Incubation | Activity after incubation (20 hours at 45Β°C), measured by colorimetric assay with Ellman's reagent (5-thio-2-nitrobenzoate anion absorbance measurement, 412 nm) in aqueous phase | Dimethyl Sulfoxide (DMSO) | 25% | 100.0 | 56.1 | 69.5 | % | Incubation: 100 mM Tris-HCl buffer, Assay: 100 mM Tris-HCl buffer | Incubation: 8, Assay: 8 | Incubation: 45Β°C, Assay: 30Β°C | ? mM 2,3-Dimercapto-1-propanol tributyrate | butyrate , Free thiols | β | Incubation: 160 rpm | Non-incubated control (in %), assay in aqueous phase | Uncertainty about whether assay really was in aqueous phase |
| Y200N | Stability - Incubation | Activity after incubation (20 hours at 45Β°C), measured by colorimetric assay with Ellman's reagent (5-thio-2-nitrobenzoate anion absorbance measurement, 412 nm) in aqueous phase | Dimethyl Sulfoxide (DMSO) | 25% | 100.0 | 56.1 | 64.1 | % | Incubation: 100 mM Tris-HCl buffer, Assay: 100 mM Tris-HCl buffer | Incubation: 8, Assay: 8 | Incubation: 45Β°C, Assay: 30Β°C | ? mM 2,3-Dimercapto-1-propanol tributyrate | butyrate , Free thiols | β | Incubation: 160 rpm | Non-incubated control (in %), assay in aqueous phase | Uncertainty about whether assay really was in aqueous phase |
| T236N | Stability - Incubation | Activity after incubation (20 hours at 45Β°C), measured by colorimetric assay with Ellman's reagent (5-thio-2-nitrobenzoate anion absorbance measurement, 412 nm) in aqueous phase | Dimethyl Sulfoxide (DMSO) | 25% | 100.0 | 56.1 | 72.8 | % | Incubation: 100 mM Tris-HCl buffer, Assay: 100 mM Tris-HCl buffer | Incubation: 8, Assay: 8 | Incubation: 45Β°C, Assay: 30Β°C | ? mM 2,3-Dimercapto-1-propanol tributyrate | butyrate , Free thiols | β | Incubation: 160 rpm | Non-incubated control (in %), assay in aqueous phase | Uncertainty about whether assay really was in aqueous phase |
| T238N | Stability - Incubation | Activity after incubation (20 hours at 45Β°C), measured by colorimetric assay with Ellman's reagent (5-thio-2-nitrobenzoate anion absorbance measurement, 412 nm) in aqueous phase | Dimethyl Sulfoxide (DMSO) | 25% | 100.0 | 56.1 | 38.9 | % | Incubation: 100 mM Tris-HCl buffer, Assay: 100 mM Tris-HCl buffer | Incubation: 8, Assay: 8 | Incubation: 45Β°C, Assay: 30Β°C | ? mM 2,3-Dimercapto-1-propanol tributyrate | butyrate , Free thiols | β | Incubation: 160 rpm | Non-incubated control (in %), assay in aqueous phase | Uncertainty about whether assay really was in aqueous phase |
| P261N | Stability - Incubation | Activity after incubation (20 hours at 45Β°C), measured by colorimetric assay with Ellman's reagent (5-thio-2-nitrobenzoate anion absorbance measurement, 412 nm) in aqueous phase | Dimethyl Sulfoxide (DMSO) | 25% | 100.0 | 56.1 | 43.6 | % | Incubation: 100 mM Tris-HCl buffer, Assay: 100 mM Tris-HCl buffer | Incubation: 8, Assay: 8 | Incubation: 45Β°C, Assay: 30Β°C | ? mM 2,3-Dimercapto-1-propanol tributyrate | butyrate , Free thiols | β | Incubation: 160 rpm | Non-incubated control (in %), assay in aqueous phase | Uncertainty about whether assay really was in aqueous phase |
Mutations in this dataset (7)
WT
Y200N
T236N
T238N
P261N
A277N
A290N
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.