Lipase B (gene CalB) from Candida antarctica
Enzyme Description
Sequence
LPSGSDPAFSQPKSVLDAGLTCQGASPSSVSKPILLVPGTGTTGPQSFDSNWIPLSTQLGYTPCWISPPPFMLNDTQVNTEYMVNAITALYAGSGNNKLPVLTWSQGGLVAQWGLTFFPSIRSKVDRLMAFAPDYKGTVLAGPLDALAVSAPSVWQQTTGSALTTALRNAGGLTQIVPTTNLYSATDEIVQPQVSNSPLDSSYLFNGKNVQAQAVCGPLFVIDHAGSLTSQFSYVVGRSALRSTTGQARSADYGITDCNPLPANDLTPEQKVAAAALLAPAAAAIVAGPKQNCEPDLMPYARPFAVGKRTCSGIVTP
Guangnan Ou et al. (2011)
β Lipases are soluble and active in glycerol carbonate as a novel biosolvent
Enzyme and Microbial Technology
Β· doi:10.1016/j.enzmictec.2011.04.011 β
Β· Activity + Stability - Conversion
▼
Database ID
UniProt: P41365 β
Sequence Annotation
Inferred - from protein name (first 25 residues from UniProt sequence removed from mature protein)
Protein Source
Commercially purchased
Experimental Data (3 measurements)
3 measurements
| Property | Assay | Solvent | Solvent Volume | Aqueous Reference | Measured Value | Units | Solution | pH | Temperature | Substrate(s) | Product(s) | Cofactor(s) | Shaking | Comments |
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| Activity + Stability - Conversion | Conversion after incubation (1 hour at 40Β°C) in the presence of organic solvent, measured by HPLC | Glycerol Carbonate | 100 (traces of water)% | 57.4 | 55.1 | % | β | β | 40Β°C | 1.21 mmol/500 Β΅L 1-Butanol, 0.83 mmol/500 Β΅L Ethyl butyrate | Butyl butyrate , Ethanol | β | 300 rpm | Classical aqueous control (in %) |
| Activity + Stability - Conversion | Conversion after incubation (1 hour at 40Β°C) in the presence of organic solvent, measured by HPLC | Dimethylformamide (DMF) | 100 (traces of water)% | 57.4 | 0.0 | % | β | β | 40Β°C | 1.21 mmol/500 Β΅L 1-Butanol, 0.83 mmol/500 Β΅L Ethyl butyrate | Butyl butyrate , Ethanol | β | 300 rpm | Classical aqueous control (in %) |
| Activity + Stability - Conversion | Conversion after incubation (1 hour at 40Β°C) in the presence of organic solvent, measured by HPLC | Acetonitrile | 100 (traces of water)% | 57.4 | 10.8 | % | β | β | 40Β°C | 1.21 mmol/500 Β΅L 1-Butanol, 0.83 mmol/500 Β΅L Ethyl butyrate | Butyl butyrate , Ethanol | β | 300 rpm | Classical aqueous control (in %) |
Visualization : Activity + Stability β Conversion
Guangnan Ou et al. (2011)
One bar per measurement. Colour = solvent, shade = solvent volume.
Hyun June Park et al. (2013)
β Prediction of the solvent affecting site and the computational design of stable Candida antarctica lipase B in a hydrophilic organic solvent
Journal of Biotechnology
Β· doi:10.1016/j.jbiotec.2012.11.006 β
Β· Stability - Incubation Stability - Half-life
▼
Database ID
UniProt: P41365 β
Sequence Annotation
Inferred (first 25 residues from UniProt sequence removed)
Protein Source
Recombinant, host yeast Pichia pastoris X-33
Experimental Data (24 measurements)
24 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 - Half-life | Half-life at 30Β°C, with remaining activity measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) | Methanol | 80% | β | β | 35.0 | hours | Incubation: ?, Assay: 50 mM sodium phosphate, 0.5% Triton X-100 | Incubation: ?, Assay: 8 | Incubation: 30Β°C, Assay: 30Β°C | 3.33 mM p-Nitrophenyl caprylate | Caprylic Acid , p-Nitrophenol | β | β | Wild-type control with organic solvent (in hours) |
| WT | Stability - Incubation | Activity after incubation (72 hours at 30Β°C), measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in aqueous phase | Methanol | 80% | 100.0 | β | 23.9 | % | Incubation: ?, Assay: 50 mM sodium phosphate, 0.5% Triton X-100 | Incubation: ?, Assay: 8 | Incubation: 30Β°C, Assay: 30Β°C | 3.33 mM p-Nitrophenyl caprylate | Caprylic Acid , p-Nitrophenol | β | β | Non-incubated control (in %), reaction in aqueous phase | Uncertainty about whether activity was measured with or without organic solvent |
| WT | Stability - Incubation | Activity after incubation (48 hours at 30Β°C), measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in aqueous phase | Methanol | 80% | 100.0 | β | 42.6 | % | Incubation: ?, Assay: 50 mM sodium phosphate, 0.5% Triton X-100 | Incubation: ?, Assay: 8 | Incubation: 30Β°C, Assay: 30Β°C | 3.33 mM p-Nitrophenyl caprylate | Caprylic Acid , p-Nitrophenol | β | β | Non-incubated control (in %), reaction in aqueous phase | Uncertainty about whether activity was measured with or without organic solvent |
| WT | Stability - Incubation | Activity after incubation (24 hours at 30Β°C), measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in aqueous phase | Methanol | 80% | 100.0 | β | 55.9 | % | Incubation: ?, Assay: 50 mM sodium phosphate, 0.5% Triton X-100 | Incubation: ?, Assay: 8 | Incubation: 30Β°C, Assay: 30Β°C | 3.33 mM p-Nitrophenyl caprylate | Caprylic Acid , p-Nitrophenol | β | β | Non-incubated control (in %), reaction in aqueous phase | Uncertainty about whether activity was measured with or without organic solvent |
| A8T | Stability - Half-life | Half-life at 30Β°C, with remaining activity measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) | Methanol | 80% | β | 35.0 | 52.0 | hours | Incubation: ?, Assay: 50 mM sodium phosphate, 0.5% Triton X-100 | Incubation: ?, Assay: 8 | Incubation: 30Β°C, Assay: 30Β°C | 3.33 mM p-Nitrophenyl caprylate | Caprylic Acid , p-Nitrophenol | β | β | Wild-type control with organic solvent (in hours) |
| T244D | Stability - Half-life | Half-life at 30Β°C, with remaining activity measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) | Methanol | 80% | β | 35.0 | 29.0 | hours | Incubation: ?, Assay: 50 mM sodium phosphate, 0.5% Triton X-100 | Incubation: ?, Assay: 8 | Incubation: 30Β°C, Assay: 30Β°C | 3.33 mM p-Nitrophenyl caprylate | Caprylic Acid , p-Nitrophenol | β | β | Wild-type control with organic solvent (in hours) |
| T245S | Stability - Half-life | Half-life at 30Β°C, with remaining activity measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) | Methanol | 80% | β | 35.0 | 59.0 | hours | Incubation: ?, Assay: 50 mM sodium phosphate, 0.5% Triton X-100 | Incubation: ?, Assay: 8 | Incubation: 30Β°C, Assay: 30Β°C | 3.33 mM p-Nitrophenyl caprylate | Caprylic Acid , p-Nitrophenol | β | β | Wild-type control with organic solvent (in hours) |
| N97Q | Stability - Half-life | Half-life at 30Β°C, with remaining activity measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) | Methanol | 80% | β | 35.0 | 52.0 | hours | Incubation: ?, Assay: 50 mM sodium phosphate, 0.5% Triton X-100 | Incubation: ?, Assay: 8 | Incubation: 30Β°C, Assay: 30Β°C | 3.33 mM p-Nitrophenyl caprylate | Caprylic Acid , p-Nitrophenol | β | β | Wild-type control with organic solvent (in hours) |
| A92E | Stability - Half-life | Half-life at 30Β°C, with remaining activity measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) | Methanol | 80% | β | 35.0 | 63.0 | hours | Incubation: ?, Assay: 50 mM sodium phosphate, 0.5% Triton X-100 | Incubation: ?, Assay: 8 | Incubation: 30Β°C, Assay: 30Β°C | 3.33 mM p-Nitrophenyl caprylate | Caprylic Acid , p-Nitrophenol | β | β | Wild-type control with organic solvent (in hours) |
| T245S | Stability - Incubation | Activity after incubation (72 hours at 30Β°C), measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in aqueous phase | Methanol | 80% | 100.0 | 23.9 | 43.4 | % | Incubation: ?, Assay: 50 mM sodium phosphate, 0.5% Triton X-100 | Incubation: ?, Assay: 8 | Incubation: 30Β°C, Assay: 30Β°C | 3.33 mM p-Nitrophenyl caprylate | Caprylic Acid , p-Nitrophenol | β | β | Non-incubated control (in %), reaction in aqueous phase | Uncertainty about whether activity was measured with or without organic solvent |
| T244D | Stability - Incubation | Activity after incubation (72 hours at 30Β°C), measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in aqueous phase | Methanol | 80% | 100.0 | 23.9 | 17.3 | % | Incubation: ?, Assay: 50 mM sodium phosphate, 0.5% Triton X-100 | Incubation: ?, Assay: 8 | Incubation: 30Β°C, Assay: 30Β°C | 3.33 mM p-Nitrophenyl caprylate | Caprylic Acid , p-Nitrophenol | β | β | Non-incubated control (in %), reaction in aqueous phase | Uncertainty about whether activity was measured with or without organic solvent |
| N97Q | Stability - Incubation | Activity after incubation (72 hours at 30Β°C), measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in aqueous phase | Methanol | 80% | 100.0 | 23.9 | 40.8 | % | Incubation: ?, Assay: 50 mM sodium phosphate, 0.5% Triton X-100 | Incubation: ?, Assay: 8 | Incubation: 30Β°C, Assay: 30Β°C | 3.33 mM p-Nitrophenyl caprylate | Caprylic Acid , p-Nitrophenol | β | β | Non-incubated control (in %), reaction in aqueous phase | Uncertainty about whether activity was measured with or without organic solvent |
| A92E | Stability - Incubation | Activity after incubation (72 hours at 30Β°C), measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in aqueous phase | Methanol | 80% | 100.0 | 23.9 | 47.4 | % | Incubation: ?, Assay: 50 mM sodium phosphate, 0.5% Triton X-100 | Incubation: ?, Assay: 8 | Incubation: 30Β°C, Assay: 30Β°C | 3.33 mM p-Nitrophenyl caprylate | Caprylic Acid , p-Nitrophenol | β | β | Non-incubated control (in %), reaction in aqueous phase | Uncertainty about whether activity was measured with or without organic solvent |
| A8T | Stability - Incubation | Activity after incubation (72 hours at 30Β°C), measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in aqueous phase | Methanol | 80% | 100.0 | 23.9 | 42.6 | % | Incubation: ?, Assay: 50 mM sodium phosphate, 0.5% Triton X-100 | Incubation: ?, Assay: 8 | Incubation: 30Β°C, Assay: 30Β°C | 3.33 mM p-Nitrophenyl caprylate | Caprylic Acid , p-Nitrophenol | β | β | Non-incubated control (in %), reaction in aqueous phase | Uncertainty about whether activity was measured with or without organic solvent |
| T245S | Stability - Incubation | Activity after incubation (48 hours at 30Β°C), measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in aqueous phase | Methanol | 80% | 100.0 | 42.6 | 58.3 | % | Incubation: ?, Assay: 50 mM sodium phosphate, 0.5% Triton X-100 | Incubation: ?, Assay: 8 | Incubation: 30Β°C, Assay: 30Β°C | 3.33 mM p-Nitrophenyl caprylate | Caprylic Acid , p-Nitrophenol | β | β | Non-incubated control (in %), reaction in aqueous phase | Uncertainty about whether activity was measured with or without organic solvent |
| T244D | Stability - Incubation | Activity after incubation (48 hours at 30Β°C), measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in aqueous phase | Methanol | 80% | 100.0 | 42.6 | 38.6 | % | Incubation: ?, Assay: 50 mM sodium phosphate, 0.5% Triton X-100 | Incubation: ?, Assay: 8 | Incubation: 30Β°C, Assay: 30Β°C | 3.33 mM p-Nitrophenyl caprylate | Caprylic Acid , p-Nitrophenol | β | β | Non-incubated control (in %), reaction in aqueous phase | Uncertainty about whether activity was measured with or without organic solvent |
| N97Q | Stability - Incubation | Activity after incubation (48 hours at 30Β°C), measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in aqueous phase | Methanol | 80% | 100.0 | 42.6 | 51.6 | % | Incubation: ?, Assay: 50 mM sodium phosphate, 0.5% Triton X-100 | Incubation: ?, Assay: 8 | Incubation: 30Β°C, Assay: 30Β°C | 3.33 mM p-Nitrophenyl caprylate | Caprylic Acid , p-Nitrophenol | β | β | Non-incubated control (in %), reaction in aqueous phase | Uncertainty about whether activity was measured with or without organic solvent |
| A92E | Stability - Incubation | Activity after incubation (48 hours at 30Β°C), measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in aqueous phase | Methanol | 80% | 100.0 | 42.6 | 58.3 | % | Incubation: ?, Assay: 50 mM sodium phosphate, 0.5% Triton X-100 | Incubation: ?, Assay: 8 | Incubation: 30Β°C, Assay: 30Β°C | 3.33 mM p-Nitrophenyl caprylate | Caprylic Acid , p-Nitrophenol | β | β | Non-incubated control (in %), reaction in aqueous phase | Uncertainty about whether activity was measured with or without organic solvent |
| A8T | Stability - Incubation | Activity after incubation (48 hours at 30Β°C), measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in aqueous phase | Methanol | 80% | 100.0 | 42.6 | 50.8 | % | Incubation: ?, Assay: 50 mM sodium phosphate, 0.5% Triton X-100 | Incubation: ?, Assay: 8 | Incubation: 30Β°C, Assay: 30Β°C | 3.33 mM p-Nitrophenyl caprylate | Caprylic Acid , p-Nitrophenol | β | β | Non-incubated control (in %), reaction in aqueous phase | Uncertainty about whether activity was measured with or without organic solvent |
| T245S | Stability - Incubation | Activity after incubation (24 hours at 30Β°C), measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in aqueous phase | Methanol | 80% | 100.0 | 55.9 | 70.0 | % | Incubation: ?, Assay: 50 mM sodium phosphate, 0.5% Triton X-100 | Incubation: ?, Assay: 8 | Incubation: 30Β°C, Assay: 30Β°C | 3.33 mM p-Nitrophenyl caprylate | Caprylic Acid , p-Nitrophenol | β | β | Non-incubated control (in %), reaction in aqueous phase | Uncertainty about whether activity was measured with or without organic solvent |
Mutations in this dataset (6) β Hyun June Park et al. (2013)
WT
A8T
A92E
N97Q
T244D
T245S
Visualization : Stability β Incubation
Hyun June Park et al. (2013)
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
Hyun June Park et al. (2013)
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.
So Yeon Hong et al. (2013)
β Activity Enhancement of Candida antarctica Lipase B by Flexibility Modulation in Helix Region Surrounding the Active Site
Applied Biochemistry and Biotechnology
Β· doi:10.1007/s12010-013-0237-8 β
Β· Stability - Incubation
▼
Database ID
UniProt: P41365 β
Sequence Annotation
Explicit - Provided PDB Accession Number (25 first residues from UniProt sequence absent from the mature protein)
Protein Source
Recombinant, host yeast Pichia pastoris X-33
Experimental Data (3 measurements)
3 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 (86 hours at 30Β°C), measured by absorbance spectrophotometry (p-nitrophenol absorbance measurment, 405 nm) in aqueous phase | Methanol | 50% | 100.0 | β | 52.6 | % | Incubation: 0.04% Triton X-100, 50 mM phosphate buffer, Assay: 9% acetonitrile, 0.04% Triton X-100, 50 mM phosphate buffer | Incubation: 6, Assay: 6 | Incubation: 30Β°C, Assay: 30Β°C | 3 mM p-Nitrophenyl caprylate | Caprylic Acid , p-Nitrophenol | β | β | Non-incubated control (in %), activity measured in aqueous phase | Uncertainty about whether activity was measured with or without organic solvent |
| V139E | Stability - Incubation | Activity after incubation (86 hours at 30Β°C), measured by absorbance spectrophotometry (p-nitrophenol absorbance measurment, 405 nm) in aqueous phase | Methanol | 50% | 100.0 | 52.6 | 44.1 | % | Incubation: 0.04% Triton X-100, 50 mM phosphate buffer, Assay: 9% acetonitrile, 0.04% Triton X-100, 50 mM phosphate buffer | Incubation: 6, Assay: 6 | Incubation: 30Β°C, Assay: 30Β°C | 3 mM p-Nitrophenyl caprylate | Caprylic Acid , p-Nitrophenol | β | β | Non-incubated control(in %), activity measured in aqueous phase | Uncertainty about whether activity was measured with or without organic solvent |
| I255E | Stability - Incubation | Activity after incubation (86 hours at 30Β°C), measured by absorbance spectrophotometry (p-nitrophenol absorbance measurment, 405 nm) in aqueous phase | Methanol | 50% | 100.0 | 52.6 | 39.0 | % | Incubation: 0.04% Triton X-100, 50 mM phosphate buffer, Assay: 9% acetonitrile, 0.04% Triton X-100, 50 mM phosphate buffer | Incubation: 6, Assay: 6 | Incubation: 30Β°C, Assay: 30Β°C | 3 mM p-Nitrophenyl caprylate | Caprylic Acid , p-Nitrophenol | 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 (3) β So Yeon Hong et al. (2013)
WT
V139E
I255E
Visualization : Stability β Incubation
So Yeon Hong et al. (2013)
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.
Marco Mangiagalli et al. (2020)
β Diverse effects of aqueous polar co-solvents on Candida antarctica lipase B
International Journal of Biological Macromolecules
Β· doi:10.1016/j.ijbiomac.2020.02.145 β
Β· Activity - Classical Stability - Incubation
▼
Database ID
UniProt: P41365 β
Sequence Annotation
Explicit - Provided PDB Accession Number (25 first residues from UniProt sequence removed from mature protein, 1 mutation)
Protein Source
Recombinant, host bacterium Escherichia coli BL21 (DE3)
Experimental Data (39 measurements)
39 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 |
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| N74S | Activity - Classical | Activity measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 410 nm) in the presence of organic solvent | Dimethyl Sulfoxide (DMSO) | 20% | 100.0 | β | 133.0 | % | 10 mM Tris-HCl buffer, 1% Triton X-100 | 8 | 25Β°C | 1 mM p-Nitrophenyl caprylate | Caprylic Acid , p-Nitrophenol | β | β | Classical aqueous control (in %) |
| N74S | Activity - Classical | Activity measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 410 nm) in the presence of organic solvent | Dimethyl Sulfoxide (DMSO) | 40% | 2.0 | β | 2.91 | mM/min | 10 mM Tris-HCl buffer, 1% Triton X-100 | 8 | 25Β°C | 2.5 mM p-Nitrophenyl caprylate | Caprylic Acid , p-Nitrophenol | β | β | Classical aqueous control (in mM/min) |
| N74S | Activity - Classical | Activity measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 410 nm) in the presence of organic solvent | Dimethyl Sulfoxide (DMSO) | 30% | 2.0 | β | 2.59 | mM/min | 10 mM Tris-HCl buffer, 1% Triton X-100 | 8 | 25Β°C | 2.5 mM p-Nitrophenyl caprylate | Caprylic Acid , p-Nitrophenol | β | β | Classical aqueous control (in mM/min) |
| N74S | Activity - Classical | Activity measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 410 nm) in the presence of organic solvent | Dimethyl Sulfoxide (DMSO) | 20% | 2.0 | β | 2.41 | mM/min | 10 mM Tris-HCl buffer, 1% Triton X-100 | 8 | 25Β°C | 2.5 mM p-Nitrophenyl caprylate | Caprylic Acid , p-Nitrophenol | β | β | Classical aqueous control (in mM/min) |
| N74S | Activity - Classical | Activity measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 410 nm) in the presence of organic solvent | Dimethyl Sulfoxide (DMSO) | 10% | 2.0 | β | 2.24 | mM/min | 10 mM Tris-HCl buffer, 1% Triton X-100 | 8 | 25Β°C | 2.5 mM p-Nitrophenyl caprylate | Caprylic Acid , p-Nitrophenol | β | β | Classical aqueous control (in mM/min) |
| N74S | Activity - Classical | Activity measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 410 nm) in the presence of organic solvent | Methanol | 40% | 2.0 | β | 1.05 | mM/min | 10 mM Tris-HCl buffer, 1% Triton X-100 | 8 | 25Β°C | 2.5 mM p-Nitrophenyl caprylate | Caprylic Acid , p-Nitrophenol | β | β | Classical aqueous control (in mM/min) |
| N74S | Activity - Classical | Activity measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 410 nm) in the presence of organic solvent | Methanol | 30% | 2.0 | β | 1.26 | mM/min | 10 mM Tris-HCl buffer, 1% Triton X-100 | 8 | 25Β°C | 2.5 mM p-Nitrophenyl caprylate | Caprylic Acid , p-Nitrophenol | β | β | Classical aqueous control (in mM/min) |
| N74S | Activity - Classical | Activity measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 410 nm) in the presence of organic solvent | Methanol | 20% | 2.0 | β | 1.44 | mM/min | 10 mM Tris-HCl buffer, 1% Triton X-100 | 8 | 25Β°C | 2.5 mM p-Nitrophenyl caprylate | Caprylic Acid , p-Nitrophenol | β | β | Classical aqueous control (in mM/min) |
| N74S | Activity - Classical | Activity measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 410 nm) in the presence of organic solvent | Methanol | 10% | 2.0 | β | 1.73 | mM/min | 10 mM Tris-HCl buffer, 1% Triton X-100 | 8 | 25Β°C | 2.5 mM p-Nitrophenyl caprylate | Caprylic Acid , p-Nitrophenol | β | β | Classical aqueous control (in mM/min) |
| N74S | Activity - Classical | Activity measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 410 nm) in the presence of organic solvent | Acetone | 25% | 1.8 | β | 0.97 | mM/min | 10 mM Tris-HCl buffer, 1% Triton X-100 | 8 | 25Β°C | 1.5 mM p-Nitrophenyl caprylate | Caprylic Acid , p-Nitrophenol | β | β | Classical aqueous control (in mM/min) |
| N74S | Activity - Classical | Activity measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 410 nm) in the presence of organic solvent | Acetone | 20% | 1.8 | β | 1.08 | mM/min | 10 mM Tris-HCl buffer, 1% Triton X-100 | 8 | 25Β°C | 1.5 mM p-Nitrophenyl caprylate | Caprylic Acid , p-Nitrophenol | β | β | Classical aqueous control (in mM/min) |
| N74S | Activity - Classical | Activity measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 410 nm) in the presence of organic solvent | Acetone | 15% | 1.8 | β | 1.18 | mM/min | 10 mM Tris-HCl buffer, 1% Triton X-100 | 8 | 25Β°C | 1.5 mM p-Nitrophenyl caprylate | Caprylic Acid , p-Nitrophenol | β | β | Classical aqueous control (in mM/min) |
| N74S | Activity - Classical | Activity measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 410 nm) in the presence of organic solvent | Acetone | 10% | 1.8 | β | 1.32 | mM/min | 10 mM Tris-HCl buffer, 1% Triton X-100 | 8 | 25Β°C | 1.5 mM p-Nitrophenyl caprylate | Caprylic Acid , p-Nitrophenol | β | β | Classical aqueous control (in mM/min) |
| N74S | Activity - Classical | Activity measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 410 nm) in the presence of organic solvent | Dimethyl Sulfoxide (DMSO) | 10% | 100.0 | β | 125.0 | % | 10 mM Tris-HCl buffer, 1% Triton X-100 | 8 | 25Β°C | 1 mM p-Nitrophenyl caprylate | Caprylic Acid , p-Nitrophenol | β | β | Classical aqueous control (in %) |
| N74S | Activity - Classical | Activity measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 410 nm) in the presence of organic solvent | Dimethyl Sulfoxide (DMSO) | 30% | 100.0 | β | 149.0 | % | 10 mM Tris-HCl buffer, 1% Triton X-100 | 8 | 25Β°C | 1 mM p-Nitrophenyl caprylate | Caprylic Acid , p-Nitrophenol | β | β | Classical aqueous control (in %) |
| N74S | Activity - Classical | Activity measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 410 nm) in the presence of organic solvent | Dimethyl Sulfoxide (DMSO) | 40% | 100.0 | β | 166.0 | % | 10 mM Tris-HCl buffer, 1% Triton X-100 | 8 | 25Β°C | 1 mM p-Nitrophenyl caprylate | Caprylic Acid , p-Nitrophenol | β | β | Classical aqueous control (in %) |
| N74S | Activity - Classical | Activity measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 410 nm) in the presence of organic solvent | Methanol | 10% | 100.0 | β | 72.0 | % | 10 mM Tris-HCl buffer, 1% Triton X-100 | 8 | 25Β°C | 1 mM p-Nitrophenyl caprylate | Caprylic Acid , p-Nitrophenol | β | β | Classical aqueous control (in %) |
| N74S | Activity - Classical | Activity measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 410 nm) in the presence of organic solvent | Methanol | 20% | 100.0 | β | 59.0 | % | 10 mM Tris-HCl buffer, 1% Triton X-100 | 8 | 25Β°C | 1 mM p-Nitrophenyl caprylate | Caprylic Acid , p-Nitrophenol | β | β | Classical aqueous control (in %) |
| N74S | Activity - Classical | Activity measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 410 nm) in the presence of organic solvent | Methanol | 30% | 100.0 | β | 42.0 | % | 10 mM Tris-HCl buffer, 1% Triton X-100 | 8 | 25Β°C | 1 mM p-Nitrophenyl caprylate | Caprylic Acid , p-Nitrophenol | β | β | Classical aqueous control (in %) |
| N74S | Activity - Classical | Activity measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 410 nm) in the presence of organic solvent | Methanol | 40% | 100.0 | β | 37.0 | % | 10 mM Tris-HCl buffer, 1% Triton X-100 | 8 | 25Β°C | 1 mM p-Nitrophenyl caprylate | Caprylic Acid , p-Nitrophenol | β | β | Classical aqueous control (in %) |
Mutations in this dataset (1) β Marco Mangiagalli et al. (2020)
N74S
Visualization : Activity β Classical
Marco Mangiagalli et al. (2020)
One bar per measurement. Colour = solvent, shade = solvent volume.
Visualization : Stability β Incubation
Marco Mangiagalli et al. (2020)
One bar per measurement. Colour = solvent, shade = solvent volume.
Marcela Robles-Machuca et al. (2025)
β Further Characterization of Lipase B from Ustilago maydis Expressed in Pichia pastoris: a Member of the Candida antarctica Lipase B-like Superfamily
Applied Biochemistry and Biotechnology
Β· doi:10.1007/s12010-024-05166-0 β
Β· Stability - Incubation
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Database ID
UniProt: P41365 β
Sequence Annotation
Explicit - Provided GenBank Accession Number
Protein Source
Recombinant, host yeast Pichia pastoris X-33
Experimental Data (10 measurements)
10 measurements
| Property | Assay | Solvent | Solvent Volume | Aqueous Reference | Measured Value | Units | Solution | pH | Temperature | Substrate(s) | Product(s) | Cofactor(s) | Shaking | Comments |
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| Stability - Incubation | Activity after incubation (2 hours at 30Β°C), measured by pH shift caused by fatty acid release (p-nitrophenol indicator) in aqueous phase | Dimethyl Sulfoxide (DMSO) | 30% | 100 | 65 | % | 2.5 mM 4-morpholinepropane- sulfonic acid buffer incubation, 2.5 mM 4-morpholinepropane- sulfonic acid buffer, 150 mM NaCl, 6 mM CaCl2, 0.5 MM sodium taurodeoxycholate, 3 mg/ml Ξ²-cyclodextrin | Incubation: 7.2, Assay: 7 | Incubation: 30Β°C, Assay: ? | 50 mM Tributyrin | 3-butyric acid , glycerol | β | Incubation: 1000 rpm | Non-incubated control (in %), assay in aqueous phase | Clear about assay in aqueous phase ("After incubation, the organic phase was separated by centrifugation at maxi- mum speed for 15 min for samples containing hydrophobic solvents, retaining only the aqueous phase where the enzyme was present. For hydrophilic solvent samples, a fraction of the solvent-buffer mixture was taken directly. In both cases, the sample was then diluted tenfold in 2.5 mmol Lβ1 MOPS buffer at pH 7") |
| Stability - Incubation | Activity after incubation (2 hours at 30Β°C), measured by pH shift caused by fatty acid release (p-nitrophenol indicator) in aqueous phase | Methanol | 30% | 100 | 24 | % | 2.5 mM 4-morpholinepropane- sulfonic acid buffer incubation, 2.5 mM 4-morpholinepropane- sulfonic acid buffer, 150 mM NaCl, 6 mM CaCl2, 0.5 MM sodium taurodeoxycholate, 3 mg/ml Ξ²-cyclodextrin | Incubation: 7.2, Assay: 7 | Incubation: 30Β°C, Assay: ? | 50 mM Tributyrin | 3-butyric acid , glycerol | β | Incubation: 1000 rpm | Non-incubated control (in %), assay in aqueous phase | Clear about assay in aqueous phase ("After incubation, the organic phase was separated by centrifugation at maxi- mum speed for 15 min for samples containing hydrophobic solvents, retaining only the aqueous phase where the enzyme was present. For hydrophilic solvent samples, a fraction of the solvent-buffer mixture was taken directly. In both cases, the sample was then diluted tenfold in 2.5 mmol Lβ1 MOPS buffer at pH 7") |
| Stability - Incubation | Activity after incubation (2 hours at 30Β°C), measured by pH shift caused by fatty acid release (p-nitrophenol indicator) in aqueous phase | Acetonitrile | 30% | 100 | 98 | % | 2.5 mM 4-morpholinepropane- sulfonic acid buffer incubation, 2.5 mM 4-morpholinepropane- sulfonic acid buffer, 150 mM NaCl, 6 mM CaCl2, 0.5 MM sodium taurodeoxycholate, 3 mg/ml Ξ²-cyclodextrin | Incubation: 7.2, Assay: 7 | Incubation: 30Β°C, Assay: ? | 50 mM Tributyrin | 3-butyric acid , glycerol | β | Incubation: 1000 rpm | Non-incubated control (in %), assay in aqueous phase | Clear about assay in aqueous phase ("After incubation, the organic phase was separated by centrifugation at maxi- mum speed for 15 min for samples containing hydrophobic solvents, retaining only the aqueous phase where the enzyme was present. For hydrophilic solvent samples, a fraction of the solvent-buffer mixture was taken directly. In both cases, the sample was then diluted tenfold in 2.5 mmol Lβ1 MOPS buffer at pH 7") |
| Stability - Incubation | Activity after incubation (2 hours at 30Β°C), measured by pH shift caused by fatty acid release (p-nitrophenol indicator) in aqueous phase | Acetone | 30% | 100 | 160 | % | 2.5 mM 4-morpholinepropane- sulfonic acid buffer incubation, 2.5 mM 4-morpholinepropane- sulfonic acid buffer, 150 mM NaCl, 6 mM CaCl2, 0.5 MM sodium taurodeoxycholate, 3 mg/ml Ξ²-cyclodextrin | Incubation: 7.2, Assay: 7 | Incubation: 30Β°C, Assay: ? | 50 mM Tributyrin | 3-butyric acid , glycerol | β | Incubation: 1000 rpm | Non-incubated control (in %), assay in aqueous phase | Clear about assay in aqueous phase ("After incubation, the organic phase was separated by centrifugation at maxi- mum speed for 15 min for samples containing hydrophobic solvents, retaining only the aqueous phase where the enzyme was present. For hydrophilic solvent samples, a fraction of the solvent-buffer mixture was taken directly. In both cases, the sample was then diluted tenfold in 2.5 mmol Lβ1 MOPS buffer at pH 7") |
| Stability - Incubation | Activity after incubation (2 hours at 30Β°C), measured by pH shift caused by fatty acid release (p-nitrophenol indicator) in aqueous phase | Tetrahydrofuran (THF) | 30% | 100 | 78 | % | 2.5 mM 4-morpholinepropane- sulfonic acid buffer incubation, 2.5 mM 4-morpholinepropane- sulfonic acid buffer, 150 mM NaCl, 6 mM CaCl2, 0.5 MM sodium taurodeoxycholate, 3 mg/ml Ξ²-cyclodextrin | Incubation: 7.2, Assay: 7 | Incubation: 30Β°C, Assay: ? | 50 mM Tributyrin | 3-butyric acid , glycerol | β | Incubation: 1000 rpm | Non-incubated control (in %), assay in aqueous phase | Clear about assay in aqueous phase ("After incubation, the organic phase was separated by centrifugation at maxi- mum speed for 15 min for samples containing hydrophobic solvents, retaining only the aqueous phase where the enzyme was present. For hydrophilic solvent samples, a fraction of the solvent-buffer mixture was taken directly. In both cases, the sample was then diluted tenfold in 2.5 mmol Lβ1 MOPS buffer at pH 7") |
| Stability - Incubation | Activity after incubation (2 hours at 30Β°C), measured by pH shift caused by fatty acid release (p-nitrophenol indicator) in aqueous phase | 1-Butanol | 30% | 100 | 34 | % | 2.5 mM 4-morpholinepropane- sulfonic acid buffer incubation, 2.5 mM 4-morpholinepropane- sulfonic acid buffer, 150 mM NaCl, 6 mM CaCl2, 0.5 MM sodium taurodeoxycholate, 3 mg/ml Ξ²-cyclodextrin | Incubation: 7.2, Assay: 7 | Incubation: 30Β°C, Assay: ? | 50 mM Tributyrin | 3-butyric acid , glycerol | β | Incubation: 1000 rpm | Non-incubated control (in %), assay in aqueous phase | Clear about assay in aqueous phase ("After incubation, the organic phase was separated by centrifugation at maxi- mum speed for 15 min for samples containing hydrophobic solvents, retaining only the aqueous phase where the enzyme was present. For hydrophilic solvent samples, a fraction of the solvent-buffer mixture was taken directly. In both cases, the sample was then diluted tenfold in 2.5 mmol Lβ1 MOPS buffer at pH 7") |
| Stability - Incubation | Activity after incubation (2 hours at 30Β°C), measured by pH shift caused by fatty acid release (p-nitrophenol indicator) in aqueous phase | tert-Butanol | 30% | 100 | 76 | % | 2.5 mM 4-morpholinepropane- sulfonic acid buffer incubation, 2.5 mM 4-morpholinepropane- sulfonic acid buffer, 150 mM NaCl, 6 mM CaCl2, 0.5 MM sodium taurodeoxycholate, 3 mg/ml Ξ²-cyclodextrin | Incubation: 7.2, Assay: 7 | Incubation: 30Β°C, Assay: ? | 50 mM Tributyrin | 3-butyric acid , glycerol | β | Incubation: 1000 rpm | Non-incubated control (in %), assay in aqueous phase | Clear about assay in aqueous phase ("After incubation, the organic phase was separated by centrifugation at maxi- mum speed for 15 min for samples containing hydrophobic solvents, retaining only the aqueous phase where the enzyme was present. For hydrophilic solvent samples, a fraction of the solvent-buffer mixture was taken directly. In both cases, the sample was then diluted tenfold in 2.5 mmol Lβ1 MOPS buffer at pH 7") |
| Stability - Incubation | Activity after incubation (2 hours at 30Β°C), measured by pH shift caused by fatty acid release (p-nitrophenol indicator) in aqueous phase | Toluene | 30% | 100 | 405 | % | 2.5 mM 4-morpholinepropane- sulfonic acid buffer incubation, 2.5 mM 4-morpholinepropane- sulfonic acid buffer, 150 mM NaCl, 6 mM CaCl2, 0.5 MM sodium taurodeoxycholate, 3 mg/ml Ξ²-cyclodextrin | Incubation: 7.2, Assay: 7 | Incubation: 30Β°C, Assay: ? | 50 mM Tributyrin | 3-butyric acid , glycerol | β | Incubation: 1000 rpm | Non-incubated control (in %), assay in aqueous phase | Clear about assay in aqueous phase ("After incubation, the organic phase was separated by centrifugation at maxi- mum speed for 15 min for samples containing hydrophobic solvents, retaining only the aqueous phase where the enzyme was present. For hydrophilic solvent samples, a fraction of the solvent-buffer mixture was taken directly. In both cases, the sample was then diluted tenfold in 2.5 mmol Lβ1 MOPS buffer at pH 7") |
| Stability - Incubation | Activity after incubation (2 hours at 30Β°C), measured by pH shift caused by fatty acid release (p-nitrophenol indicator) in aqueous phase | n-Hexane | 30% | 100 | 345 | % | 2.5 mM 4-morpholinepropane- sulfonic acid buffer incubation, 2.5 mM 4-morpholinepropane- sulfonic acid buffer, 150 mM NaCl, 6 mM CaCl2, 0.5 MM sodium taurodeoxycholate, 3 mg/ml Ξ²-cyclodextrin | Incubation: 7.2, Assay: 7 | Incubation: 30Β°C, Assay: ? | 50 mM Tributyrin | 3-butyric acid , glycerol | β | Incubation: 1000 rpm | Non-incubated control (in %), assay in aqueous phase | Clear about assay in aqueous phase ("After incubation, the organic phase was separated by centrifugation at maxi- mum speed for 15 min for samples containing hydrophobic solvents, retaining only the aqueous phase where the enzyme was present. For hydrophilic solvent samples, a fraction of the solvent-buffer mixture was taken directly. In both cases, the sample was then diluted tenfold in 2.5 mmol Lβ1 MOPS buffer at pH 7") |
| Stability - Incubation | Activity after incubation (2 hours at 30Β°C), measured by pH shift caused by fatty acid release (p-nitrophenol indicator) in aqueous phase | Isooctane | 30% | 100 | 338 | % | 2.5 mM 4-morpholinepropane- sulfonic acid buffer incubation, 2.5 mM 4-morpholinepropane- sulfonic acid buffer, 150 mM NaCl, 6 mM CaCl2, 0.5 MM sodium taurodeoxycholate, 3 mg/ml Ξ²-cyclodextrin | Incubation: 7.2, Assay: 7 | Incubation: 30Β°C, Assay: ? | 50 mM Tributyrin | 3-butyric acid , glycerol | β | Incubation: 1000 rpm | Non-incubated control (in %), assay in aqueous phase | Clear about assay in aqueous phase ("After incubation, the organic phase was separated by centrifugation at maxi- mum speed for 15 min for samples containing hydrophobic solvents, retaining only the aqueous phase where the enzyme was present. For hydrophilic solvent samples, a fraction of the solvent-buffer mixture was taken directly. In both cases, the sample was then diluted tenfold in 2.5 mmol Lβ1 MOPS buffer at pH 7") |
Visualization : Stability β Incubation
Marcela Robles-Machuca et al. (2025)
One bar per measurement. Colour = solvent, shade = solvent volume.