Lipase A (gene lipA) from Bacillus subtilis

Enzyme Description

Extremophile
No
EC Number

Sequence

Length: 181 amino acids
AEHNPVVMVHGIGGASFNFAGIKSYLVSQGWSRDKLYAVDFWDKTGTNYNNGPVLSRFVQKVLDETGAKKVDIVAHSMGGANTLYYIKNLDGGNKVANVVTLGGANRLTTGKALPGTDPNQKILYTSIYSSADMIVMNYLSRLDGARNVQIHGVGHIGLLYSSQVNSLIKEGLNGGGQNTN
Manfred T. Reetz et al. (2010) β€” Increasing the stability of an enzyme toward hostile organic solvents by directed evolution based on iterative saturation mutagenesis using the B-FIT method
Chemical Communications  Β· doi:10.1039/c0cc02657c β†—  Β· Activity - Classical Stability - Half-life
105 measurements
Database ID
UniProt: P37957 β†—
Sequence Annotation
Inferred - from protein (31 first residues from Uniprot sequence absent from the mature protein)
Protein Source
Recombinant, host bacterium Escherichia coli BL21 (DE3)

Experimental Data (105 measurements)

105 measurements β€” page 3 of 6
Mutation Property Assay Solvent Solvent Volume Aqueous ReferenceWT Reference Measured Value Units Solution pH Temperature Substrate(s) Product(s) Cofactor(s) Shaking Comments
M134D Activity - Classical Activity measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement) in the presence of organic solvent Dimethyl Sulfoxide (DMSO) 10% 100 75 88 % ? ? 30Β°C ? mM p-Nitrophenyl caprylate Caprylic Acid , p-Nitrophenol β€” β€” Classical aqueous control (in %)
M134D Activity - Classical Activity measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement) in the presence of organic solvent Dimethyl Sulfoxide (DMSO) 20% 100 58 78 % ? ? 30Β°C ? mM p-Nitrophenyl caprylate Caprylic Acid , p-Nitrophenol β€” β€” Classical aqueous control (in %)
M134D Activity - Classical Activity measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement) in the presence of organic solvent Dimethyl Sulfoxide (DMSO) 30% 100 56 76 % ? ? 30Β°C ? mM p-Nitrophenyl caprylate Caprylic Acid , p-Nitrophenol β€” β€” Classical aqueous control (in %)
M134D Activity - Classical Activity measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement) in the presence of organic solvent Dimethyl Sulfoxide (DMSO) 40% 100 54 74 % ? ? 30Β°C ? mM p-Nitrophenyl caprylate Caprylic Acid , p-Nitrophenol β€” β€” Classical aqueous control (in %)
M134D Activity - Classical Activity measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement) in the presence of organic solvent Dimethyl Sulfoxide (DMSO) 50% 100 50 70 % ? ? 30Β°C ? mM p-Nitrophenyl caprylate Caprylic Acid , p-Nitrophenol β€” β€” Classical aqueous control (in %)
M134D/I157M Activity - Classical Activity measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement) in the presence of organic solvent Dimethyl Sulfoxide (DMSO) 10% 100 75 89 % ? ? 30Β°C ? mM p-Nitrophenyl caprylate Caprylic Acid , p-Nitrophenol β€” β€” Classical aqueous control (in %)
M134D/I157M Activity - Classical Activity measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement) in the presence of organic solvent Dimethyl Sulfoxide (DMSO) 20% 100 58 80 % ? ? 30Β°C ? mM p-Nitrophenyl caprylate Caprylic Acid , p-Nitrophenol β€” β€” Classical aqueous control (in %)
M134D/I157M Activity - Classical Activity measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement) in the presence of organic solvent Dimethyl Sulfoxide (DMSO) 30% 100 56 80 % ? ? 30Β°C ? mM p-Nitrophenyl caprylate Caprylic Acid , p-Nitrophenol β€” β€” Classical aqueous control (in %)
M134D/I157M Activity - Classical Activity measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement) in the presence of organic solvent Dimethyl Sulfoxide (DMSO) 40% 100 54 76 % ? ? 30Β°C ? mM p-Nitrophenyl caprylate Caprylic Acid , p-Nitrophenol β€” β€” Classical aqueous control (in %)
M134D/I157M Activity - Classical Activity measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement) in the presence of organic solvent Dimethyl Sulfoxide (DMSO) 50% 100 50 73 % ? ? 30Β°C ? mM p-Nitrophenyl caprylate Caprylic Acid , p-Nitrophenol β€” β€” Classical aqueous control (in %)
M134D/Y139C/I157M Activity - Classical Activity measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement) in the presence of organic solvent Dimethyl Sulfoxide (DMSO) 10% 100 75 92 % ? ? 30Β°C ? mM p-Nitrophenyl caprylate Caprylic Acid , p-Nitrophenol β€” β€” Classical aqueous control (in %)
M134D/Y139C/I157M Activity - Classical Activity measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement) in the presence of organic solvent Dimethyl Sulfoxide (DMSO) 20% 100 58 88 % ? ? 30Β°C ? mM p-Nitrophenyl caprylate Caprylic Acid , p-Nitrophenol β€” β€” Classical aqueous control (in %)
M134D/Y139C/I157M Activity - Classical Activity measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement) in the presence of organic solvent Dimethyl Sulfoxide (DMSO) 30% 100 56 84 % ? ? 30Β°C ? mM p-Nitrophenyl caprylate Caprylic Acid , p-Nitrophenol β€” β€” Classical aqueous control (in %)
M134D/Y139C/I157M Activity - Classical Activity measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement) in the presence of organic solvent Dimethyl Sulfoxide (DMSO) 40% 100 54 82 % ? ? 30Β°C ? mM p-Nitrophenyl caprylate Caprylic Acid , p-Nitrophenol β€” β€” Classical aqueous control (in %)
M134D/Y139C/I157M Activity - Classical Activity measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement) in the presence of organic solvent Dimethyl Sulfoxide (DMSO) 50% 100 50 78 % ? ? 30Β°C ? mM p-Nitrophenyl caprylate Caprylic Acid , p-Nitrophenol β€” β€” Classical aqueous control (in %)
K112D/M134D/Y139C/I157M Activity - Classical Activity measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement) in the presence of organic solvent Dimethyl Sulfoxide (DMSO) 10% 100 75 94 % ? ? 30Β°C ? mM p-Nitrophenyl caprylate Caprylic Acid , p-Nitrophenol β€” β€” Classical aqueous control (in %)
K112D/M134D/Y139C/I157M Activity - Classical Activity measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement) in the presence of organic solvent Dimethyl Sulfoxide (DMSO) 20% 100 58 92 % ? ? 30Β°C ? mM p-Nitrophenyl caprylate Caprylic Acid , p-Nitrophenol β€” β€” Classical aqueous control (in %)
K112D/M134D/Y139C/I157M Activity - Classical Activity measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement) in the presence of organic solvent Dimethyl Sulfoxide (DMSO) 30% 100 56 90 % ? ? 30Β°C ? mM p-Nitrophenyl caprylate Caprylic Acid , p-Nitrophenol β€” β€” Classical aqueous control (in %)
K112D/M134D/Y139C/I157M Activity - Classical Activity measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement) in the presence of organic solvent Dimethyl Sulfoxide (DMSO) 40% 100 54 85 % ? ? 30Β°C ? mM p-Nitrophenyl caprylate Caprylic Acid , p-Nitrophenol β€” β€” Classical aqueous control (in %)
K112D/M134D/Y139C/I157M Activity - Classical Activity measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement) in the presence of organic solvent Dimethyl Sulfoxide (DMSO) 50% 100 50 81 % ? ? 30Β°C ? mM p-Nitrophenyl caprylate Caprylic Acid , p-Nitrophenol β€” β€” Classical aqueous control (in %)
1 2 3 4 … 6

Mutations in this dataset (6) β€” Manfred T. Reetz et al. (2010)

WT R33G/K112D/M134D/Y139C/I157M K112D/M134D/Y139C/I157M M134D M134D/I157M M134D/Y139C/I157M

Visualization : Activity β€” Classical

Manfred T. Reetz et al. (2010)

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.

Visualization : Stability β€” Half-life

Manfred T. Reetz et al. (2010)

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.

Md. Zahid Kamal et al. (2013) β€” Lipase in aqueous-polar organic solvents: Activity, structure, and stability
Protein Science  Β· doi:10.1002/pro.2271 β†—  Β· Activity - Classical Activity - Michaelis-Menten Stability - Tm
124 measurements
Haiyang Cui et al. (2020) β€” How to Engineer Organic Solvent Resistant Enzymes: Insights from Combined Molecular Dynamics and Directed Evolution Study
ChemCatChem  Β· doi:10.1002/cctc.202000422 β†—  Β· Activity + Stability - Incubation
570 measurements
Kim N. Ingenbosch et al. (2021) β€” Effect of Organic Solvents on the Structure and Activity of a Minimal Lipase
The Journal of Organic Chemistry  Β· doi:10.1021/acs.joc.1c01136 β†—  Β· Activity - Classical Activity - Michaelis-Menten
39 measurements
Haiyang Cui et al. (2021) β€” CompassR Yields Highly Organic-Solvent-Tolerant Enzymes through Recombination of Compatible Substitutions
Chemistry–A European Journal  Β· doi:10.1002/chem.202004471 β†—  Β· Activity + Stability - Incubation
235 measurements
Kyoungseon Min et al. (2021) β€” Improving the organic solvent resistance of lipase a from Bacillus subtilis in water–ethanol solvent through rational surface engineering
Bioresource Technology  Β· doi:10.1016/j.biortech.2021.125394 β†—  Β· Activity - Classical Stability - Incubation
39 measurements
Haiyang Cui et al. (2021) β€” Less Unfavorable Salt Bridges on the Enzyme Surface Result in More Organic Cosolvent Resistance
Angewandte Chemie International Edition  Β· doi:10.1002/anie.202101642 β†—  Β· Activity - Michaelis-Menten Stability - Incubation
1353 measurements
Haiyang Cui et al. (2022) β€” Polar Substitutions on the Surface of a Lipase Substantially Improve Tolerance in Organic Solvents
ChemSusChem  Β· doi:10.1002/cssc.202102551 β†—  Β· Activity + Stability - Incubation
4880 measurements

Structure

Drag to rotate Β· Scroll to zoom Β· Right-click drag to pan Β· Powered by Mol*