Lipase 42 from Geobacillus sp.
Enzyme Description
Sequence
MKCCRIMFVLLGLWFVFGLSVPGGRTEAASLRANDAPIVLLHGFTGWGREEMFGFKYWGGVRGDIEQWLNDNGYRTYTLAVGPLSSNWDRACEAYAQLVGGTVDYGAAHAAKHGHARFGRTYPGLLPELKRGGRIHIIAHSQGGQTARMLVSLLENGSQEEREYAKAHNVVLSPLFEGGHHFVLSVTTIATPHDGTTLVNMVDFTDRFFDLQKAVLEAAAVASNVPYTSQVYDFKLDQWGLRRQPGESFDHYFERLKRSPVWTSTDTARYDLSVSGAEKLNQWVQASPNTYYLSFSTERTYRGALTGNHYPELGMNAFSAVVCAPFLGSYRNPTLGIDDRWLENDGIVNTVSMNGPKRGSSDRIVPYDGTLKKGVWNDMGTYNVDHLEIIGVDPNPSFDIRAFYLRLAEQLASLRP
Ummie Umaiera Mohd Johan et al. (2025)
β Methanol response of V171S lipase from Geobacillus sp. 42 explored via experimental and in silico approaches
AMB Express
Β· doi:10.1186/s13568-025-01966-7 β
Β· Stability - Incubation
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Database ID
UniProt: Q5U780 β
Sequence Annotation
Inferred - from protein name
Protein Source
Recombinant, host bacterium Escherichia coli BL21(DE3) pLysS
Experimental Data (26 measurements)
26 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 | Stability - Incubation | Activity after incubation (30 minutes at 40Β°C), measured by absorbance spectrophotometry (Kwan-Rhee method, colorimetric assay with copper reacting wit the free fatty acid products, 715 nm) in aqueous phase | Methanol | 90% | 100.0 | β | 5.2 | % | Incubation: ?, Assay: 100 mM Tris-HCl buffer, 0.1 MM CaCl2 | Incubation: ?, Assay: 8 | Incubation: 40Β°C, Assay: 70Β°C | 37% Olive oil | free fatty acids , glycerol | β | Incubation: 200 rpm, Assay: 200 rpm | Non-incubated control (in %), assay in aqueous phase | Uncertainty about whether assay really was in aqueous phase |
| WT | Stability - Incubation | Activity after incubation (30 minutes at 40Β°C), measured by absorbance spectrophotometry (Kwan-Rhee method, colorimetric assay with copper reacting wit the free fatty acid products, 715 nm) in aqueous phase | Methanol | 75% | 100.0 | β | 3.2 | % | Incubation: ?, Assay: 100 mM Tris-HCl buffer, 0.1 MM CaCl2 | Incubation: ?, Assay: 8 | Incubation: 40Β°C, Assay: 70Β°C | 37% Olive oil | free fatty acids , glycerol | β | Incubation: 200 rpm, Assay: 200 rpm | Non-incubated control (in %), assay in aqueous phase | Uncertainty about whether assay really was in aqueous phase |
| WT | Stability - Incubation | Activity after incubation (30 minutes at 40Β°C), measured by absorbance spectrophotometry (Kwan-Rhee method, colorimetric assay with copper reacting wit the free fatty acid products, 715 nm) in aqueous phase | Methanol | 60% | 100.0 | β | 5.2 | % | Incubation: ?, Assay: 100 mM Tris-HCl buffer, 0.1 MM CaCl2 | Incubation: ?, Assay: 8 | Incubation: 40Β°C, Assay: 70Β°C | 37% Olive oil | free fatty acids , glycerol | β | Incubation: 200 rpm, Assay: 200 rpm | Non-incubated control (in %), assay in aqueous phase | Uncertainty about whether assay really was in aqueous phase |
| WT | Stability - Incubation | Activity after incubation (30 minutes at 40Β°C), measured by absorbance spectrophotometry (Kwan-Rhee method, colorimetric assay with copper reacting wit the free fatty acid products, 715 nm) in aqueous phase | Methanol | 45% | 100.0 | β | 83.1 | % | Incubation: ?, Assay: 100 mM Tris-HCl buffer, 0.1 MM CaCl2 | Incubation: ?, Assay: 8 | Incubation: 40Β°C, Assay: 70Β°C | 37% Olive oil | free fatty acids , glycerol | β | Incubation: 200 rpm, Assay: 200 rpm | Non-incubated control (in %), assay in aqueous phase | Uncertainty about whether assay really was in aqueous phase |
| WT | Stability - Incubation | Activity after incubation (30 minutes at 40Β°C), measured by absorbance spectrophotometry (Kwan-Rhee method, colorimetric assay with copper reacting wit the free fatty acid products, 715 nm) in aqueous phase | Methanol | 30% | 100.0 | β | 76.3 | % | Incubation: ?, Assay: 100 mM Tris-HCl buffer, 0.1 MM CaCl2 | Incubation: ?, Assay: 8 | Incubation: 40Β°C, Assay: 70Β°C | 37% Olive oil | free fatty acids , glycerol | β | Incubation: 200 rpm, Assay: 200 rpm | Non-incubated control (in %), assay in aqueous phase | Uncertainty about whether assay really was in aqueous phase |
| WT | Stability - Incubation | Activity after incubation (30 minutes at 40Β°C), measured by absorbance spectrophotometry (Kwan-Rhee method, colorimetric assay with copper reacting wit the free fatty acid products, 715 nm) in aqueous phase | Methanol | 15% | 100.0 | β | 81.8 | % | Incubation: ?, Assay: 100 mM Tris-HCl buffer, 0.1 MM CaCl2 | Incubation: ?, Assay: 8 | Incubation: 40Β°C, Assay: 70Β°C | 37% Olive oil | free fatty acids , glycerol | β | Incubation: 200 rpm, Assay: 200 rpm | Non-incubated control (in %), assay in aqueous phase | Uncertainty about whether assay really was in aqueous phase |
| V171S | Stability - Incubation | Activity after incubation (30 minutes at 65Β°C), measured by absorbance spectrophotometry (Kwan-Rhee method, colorimetric assay with copper reacting wit the free fatty acid products, 715 nm) in aqueous phase | n-Hexadecane | 25% | 100.0 | β | 82.3 | % | Incubation: ?, Assay: 100 mM Tris-HCl buffer, 0.1 MM CaCl2 | Incubation: ?, Assay: 8 | Incubation: 65Β°C, Assay: 70Β°C | 37% Olive oil | free fatty acids , glycerol | β | Incubation: 200 rpm | Non-incubated control (in %), assay in aqueous phase | Uncertainty about whether assay really was in aqueous phase |
| V171S | Stability - Incubation | Activity after incubation (30 minutes at 40Β°C), measured by absorbance spectrophotometry (Kwan-Rhee method, colorimetric assay with copper reacting wit the free fatty acid products, 715 nm) in aqueous phase | Methanol | 90% | 100.0 | 5.2 | 5.2 | % | Incubation: ?, Assay: 100 mM Tris-HCl buffer, 0.1 MM CaCl2 | Incubation: ?, Assay: 8 | Incubation: 40Β°C, Assay: 70Β°C | 37% Olive oil | free fatty acids , glycerol | β | Incubation: 200 rpm, Assay: 200 rpm | Non-incubated control (in %), assay in aqueous phase | Uncertainty about whether assay really was in aqueous phase |
| V171S | Stability - Incubation | Activity after incubation (30 minutes at 40Β°C), measured by absorbance spectrophotometry (Kwan-Rhee method, colorimetric assay with copper reacting wit the free fatty acid products, 715 nm) in aqueous phase | Methanol | 75% | 100.0 | 3.2 | 5.2 | % | Incubation: ?, Assay: 100 mM Tris-HCl buffer, 0.1 MM CaCl2 | Incubation: ?, Assay: 8 | Incubation: 40Β°C, Assay: 70Β°C | 37% Olive oil | free fatty acids , glycerol | β | Incubation: 200 rpm, Assay: 200 rpm | Non-incubated control (in %), assay in aqueous phase | Uncertainty about whether assay really was in aqueous phase |
| V171S | Stability - Incubation | Activity after incubation (30 minutes at 40Β°C), measured by absorbance spectrophotometry (Kwan-Rhee method, colorimetric assay with copper reacting wit the free fatty acid products, 715 nm) in aqueous phase | Methanol | 60% | 100.0 | 5.2 | 11.0 | % | Incubation: ?, Assay: 100 mM Tris-HCl buffer, 0.1 MM CaCl2 | Incubation: ?, Assay: 8 | Incubation: 40Β°C, Assay: 70Β°C | 37% Olive oil | free fatty acids , glycerol | β | Incubation: 200 rpm, Assay: 200 rpm | Non-incubated control (in %), assay in aqueous phase | Uncertainty about whether assay really was in aqueous phase |
| V171S | Stability - Incubation | Activity after incubation (30 minutes at 40Β°C), measured by absorbance spectrophotometry (Kwan-Rhee method, colorimetric assay with copper reacting wit the free fatty acid products, 715 nm) in aqueous phase | Methanol | 45% | 100.0 | 83.1 | 122.6 | % | Incubation: ?, Assay: 100 mM Tris-HCl buffer, 0.1 MM CaCl2 | Incubation: ?, Assay: 8 | Incubation: 40Β°C, Assay: 70Β°C | 37% Olive oil | free fatty acids , glycerol | β | Incubation: 200 rpm, Assay: 200 rpm | Non-incubated control (in %), assay in aqueous phase | Uncertainty about whether assay really was in aqueous phase |
| V171S | Stability - Incubation | Activity after incubation (30 minutes at 40Β°C), measured by absorbance spectrophotometry (Kwan-Rhee method, colorimetric assay with copper reacting wit the free fatty acid products, 715 nm) in aqueous phase | Methanol | 30% | 100.0 | 76.3 | 112.6 | % | Incubation: ?, Assay: 100 mM Tris-HCl buffer, 0.1 MM CaCl2 | Incubation: ?, Assay: 8 | Incubation: 40Β°C, Assay: 70Β°C | 37% Olive oil | free fatty acids , glycerol | β | Incubation: 200 rpm, Assay: 200 rpm | Non-incubated control (in %), assay in aqueous phase | Uncertainty about whether assay really was in aqueous phase |
| V171S | Stability - Incubation | Activity after incubation (30 minutes at 40Β°C), measured by absorbance spectrophotometry (Kwan-Rhee method, colorimetric assay with copper reacting wit the free fatty acid products, 715 nm) in aqueous phase | Methanol | 15% | 100.0 | 81.8 | 101.9 | % | Incubation: ?, Assay: 100 mM Tris-HCl buffer, 0.1 MM CaCl2 | Incubation: ?, Assay: 8 | Incubation: 40Β°C, Assay: 70Β°C | 37% Olive oil | free fatty acids , glycerol | β | Incubation: 200 rpm, Assay: 200 rpm | Non-incubated control (in %), assay in aqueous phase | Uncertainty about whether assay really was in aqueous phase |
| V171S | Stability - Incubation | Activity after incubation (30 minutes at 65Β°C), measured by absorbance spectrophotometry (Kwan-Rhee method, colorimetric assay with copper reacting wit the free fatty acid products, 715 nm) in aqueous phase | Methanol | 25% | 100.0 | β | 71.9 | % | Incubation: ?, Assay: 100 mM Tris-HCl buffer, 0.1 MM CaCl2 | Incubation: ?, Assay: 8 | Incubation: 65Β°C, Assay: 70Β°C | 37% Olive oil | free fatty acids , glycerol | β | Incubation: 200 rpm | Non-incubated control (in %), assay in aqueous phase | Uncertainty about whether assay really was in aqueous phase |
| V171S | Stability - Incubation | Activity after incubation (30 minutes at 65Β°C), measured by absorbance spectrophotometry (Kwan-Rhee method, colorimetric assay with copper reacting wit the free fatty acid products, 715 nm) in aqueous phase | n-Dodecane | 25% | 100.0 | β | 93.6 | % | Incubation: ?, Assay: 100 mM Tris-HCl buffer, 0.1 MM CaCl2 | Incubation: ?, Assay: 8 | Incubation: 65Β°C, Assay: 70Β°C | 37% Olive oil | free fatty acids , glycerol | β | Incubation: 200 rpm | Non-incubated control (in %), assay in aqueous phase | Uncertainty about whether assay really was in aqueous phase |
| V171S | Stability - Incubation | Activity after incubation (30 minutes at 65Β°C), measured by absorbance spectrophotometry (Kwan-Rhee method, colorimetric assay with copper reacting wit the free fatty acid products, 715 nm) in aqueous phase | Isooctane | 25% | 100.0 | β | 74.1 | % | Incubation: ?, Assay: 100 mM Tris-HCl buffer, 0.1 MM CaCl2 | Incubation: ?, Assay: 8 | Incubation: 65Β°C, Assay: 70Β°C | 37% Olive oil | free fatty acids , glycerol | β | Incubation: 200 rpm | Non-incubated control (in %), assay in aqueous phase | Uncertainty about whether assay really was in aqueous phase |
| V171S | Stability - Incubation | Activity after incubation (30 minutes at 65Β°C), measured by absorbance spectrophotometry (Kwan-Rhee method, colorimetric assay with copper reacting wit the free fatty acid products, 715 nm) in aqueous phase | n-Heptane | 25% | 100.0 | β | 71.4 | % | Incubation: ?, Assay: 100 mM Tris-HCl buffer, 0.1 MM CaCl2 | Incubation: ?, Assay: 8 | Incubation: 65Β°C, Assay: 70Β°C | 37% Olive oil | free fatty acids , glycerol | β | Incubation: 200 rpm | Non-incubated control (in %), assay in aqueous phase | Uncertainty about whether assay really was in aqueous phase |
| V171S | Stability - Incubation | Activity after incubation (30 minutes at 65Β°C), measured by absorbance spectrophotometry (Kwan-Rhee method, colorimetric assay with copper reacting wit the free fatty acid products, 715 nm) in aqueous phase | n-Hexane | 25% | 100.0 | β | 83.3 | % | Incubation: ?, Assay: 100 mM Tris-HCl buffer, 0.1 MM CaCl2 | Incubation: ?, Assay: 8 | Incubation: 65Β°C, Assay: 70Β°C | 37% Olive oil | free fatty acids , glycerol | β | Incubation: 200 rpm | Non-incubated control (in %), assay in aqueous phase | Uncertainty about whether assay really was in aqueous phase |
| V171S | Stability - Incubation | Activity after incubation (30 minutes at 65Β°C), measured by absorbance spectrophotometry (Kwan-Rhee method, colorimetric assay with copper reacting wit the free fatty acid products, 715 nm) in aqueous phase | 1-Octanol | 25% | 100.0 | β | 86.1 | % | Incubation: ?, Assay: 100 mM Tris-HCl buffer, 0.1 MM CaCl2 | Incubation: ?, Assay: 8 | Incubation: 65Β°C, Assay: 70Β°C | 37% Olive oil | free fatty acids , glycerol | β | Incubation: 200 rpm | Non-incubated control (in %), assay in aqueous phase | Uncertainty about whether assay really was in aqueous phase |
| V171S | Stability - Incubation | Activity after incubation (30 minutes at 65Β°C), measured by absorbance spectrophotometry (Kwan-Rhee method, colorimetric assay with copper reacting wit the free fatty acid products, 715 nm) in aqueous phase | 1-Heptanol | 25% | 100.0 | β | 106.8 | % | Incubation: ?, Assay: 100 mM Tris-HCl buffer, 0.1 MM CaCl2 | Incubation: ?, Assay: 8 | Incubation: 65Β°C, Assay: 70Β°C | 37% Olive oil | free fatty acids , glycerol | β | Incubation: 200 rpm | Non-incubated control (in %), assay in aqueous phase | Uncertainty about whether assay really was in aqueous phase |
Mutations in this dataset (2)
WT
V171S
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.