Deglycosylated lipase BTL2 from Bacillus thermocatenulatus

Enzyme Description

Species
Bacillus thermocatenulatus β†—
Extremophile
No but "thermostable" enzyme
EC Number
3.1.1.3 β†—

Sequence

Length: 389 amino acids
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
6 measurements ▼
Database ID
PDB: 2W22 β†—
Sequence Annotation
Explicit - Provided PDB Accession Number (1 mutation)
Protein Source
Recombinant, host yeast Saccharomyces cerevisiae YPH499

Experimental Data (6 measurements)

6 measurements
Mutation Property Assay Solvent Solvent Volume Aqueous ReferenceWT Reference Measured Value Units Solution pH Temperature Substrate(s) Product(s) Cofactor(s) Shaking Comments
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 β€” 57.1 % 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
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 β€” 70.9 % 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
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 β€” 60.9 % 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
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 β€” 45.0 % 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 β€” 46.8 % 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
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 β€” 63.6 % 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 (6)

Y200N T236N T238N P261N A277N A290N

Visualization : Stability β€” Incubation

One bar per measurement. Colour = solvent, shade = solvent volume.

Structure

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