Lipase (gene ELBn12) from Enterobacter sp. Bn12
Enzyme Description
Sequence
MSTSLKYPIVLVHGLLGFDKIGGIYPYFYGIKEALEKAGAKVYIATLSALNSNEMRGEQLLEFVRKVQAETGAAKVNLIGHSQGPLACRYVAATHPDLIASVTSVNGVNHGSEVADLVRLALKPGRLPESIANAALSAFGQLLSALAGEPRLPQSGVDALDALTSEGVAAFNKKYPQGLPAEWGGQGKELDNGVYYYSWSGIIDYNPLHQGMNNLDPLHVAMLAFSILFTNERFQNDGLVGRYSSHLGKVIGSDYSMDHVDAINQLAGVVANNTDPVKLYVDHLARLQAKGL
Parisa Farrokh et al. (2014)
β Cloning and characterization of newly isolated lipase from Enterobacter sp. Bn12
Brazilian Journal of Microbiology
Β· doi:10.1590/s1517-83822014000200042 β
Β· Stability - Incubation
▼
Database ID
UniProt: K4K3B3 β
Sequence Annotation
Explicit - Provided
Protein Source
Recombinant, host bacterium Escherichia coli BL21 (DE3) pLysS
Experimental Data (6 measurements)
6 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 (1 hour at 30Β°C), measured by pH-stat activity assay in aqueous phase | Acetone | 50% | 100.0 | 119.59 | % | Incubation: 20 mM Tris-HCl,, Assay: 2% gum arabic, deionized water, 0.1M NaOH for titration | Incubation: 8, Assay: ? | Incubation: 30Β°C, Assay: ? | 2 mg/ml Triglyceride | free fatty acids , glycerol | β | β | Non-incubated control (in %), assay measured in aqueous phase | High uncertainty about whether assay in aqueous phase or in the presence of organic solvent, uncertainty about control |
| Stability - Incubation | Activity after incubation (1 hour at 30Β°C), measured by pH-stat activity assay in aqueous phase | Ethanol | 50% | 100.0 | 112.99 | % | Incubation: 20 mM Tris-HCl,, Assay: 2% gum arabic, deionized water, 0.1M NaOH for titration | Incubation: 8, Assay: ? | Incubation: 30Β°C, Assay: ? | 2 mg/ml Triglyceride | free fatty acids , glycerol | β | β | Non-incubated control (in %), assay measured in aqueous phase | High uncertainty about whether assay in aqueous phase or in the presence of organic solvent, uncertainty about control |
| Stability - Incubation | Activity after incubation (1 hour at 30Β°C), measured by pH-stat activity assay in aqueous phase | Methanol | 50% | 100.0 | 105.34 | % | Incubation: 20 mM Tris-HCl,, Assay: 2% gum arabic, deionized water, 0.1M NaOH for titration | Incubation: 8, Assay: ? | Incubation: 30Β°C, Assay: ? | 2 mg/ml Triglyceride | free fatty acids , glycerol | β | β | Non-incubated control (in %), assay measured in aqueous phase | High uncertainty about whether assay in aqueous phase or in the presence of organic solvent, uncertainty about control |
| Stability - Incubation | Activity after incubation (1 hour at 30Β°C), measured by pH-stat activity assay in aqueous phase | Glycerol | 50% | 100.0 | 111.63 | % | Incubation: 20 mM Tris-HCl,, Assay: 2% gum arabic, deionized water, 0.1M NaOH for titration | Incubation: 8, Assay: ? | Incubation: 30Β°C, Assay: ? | 2 mg/ml Triglyceride | free fatty acids , glycerol | β | β | Non-incubated control (in %), assay measured in aqueous phase | High uncertainty about whether assay in aqueous phase or in the presence of organic solvent, uncertainty about control |
| Stability - Incubation | Activity after incubation (1 hour at 30Β°C), measured by pH-stat activity assay in aqueous phase | Isopropanol | 50% | 100.0 | 115.71 | % | Incubation: 20 mM Tris-HCl,, Assay: 2% gum arabic, deionized water, 0.1M NaOH for titration | Incubation: 8, Assay: ? | Incubation: 30Β°C, Assay: ? | 2 mg/ml Triglyceride | free fatty acids , glycerol | β | β | Non-incubated control (in %), assay measured in aqueous phase | High uncertainty about whether assay in aqueous phase or in the presence of organic solvent, uncertainty about control |
| Stability - Incubation | Activity after incubation (1 hour at 30Β°C), measured by pH-stat activity assay in aqueous phase | n-Hexane | 50% | 100.0 | 120.22 | % | Incubation: 20 mM Tris-HCl,, Assay: 2% gum arabic, deionized water, 0.1M NaOH for titration | Incubation: 8, Assay: ? | Incubation: 30Β°C, Assay: ? | 2 mg/ml Triglyceride | free fatty acids , glycerol | β | β | Non-incubated control (in %), assay measured in aqueous phase | High uncertainty about whether assay in aqueous phase or in the presence of organic solvent, uncertainty about control |
Visualization : Stability β Incubation
Parisa Farrokh et al. (2014)
One bar per measurement. Colour = solvent, shade = solvent volume.
Parisa Farrokh et al. (2017)
β Role of Q177A and K173A/Q177A substitutions in thermostability and activity of the ELBn12 lipase
Biotechnology and Applied Biochemistry
Β· doi:10.1002/bab.1576 β
Β· Stability - Incubation
▼
Database ID
UniProt: K4K3B3 β
Sequence Annotation
Explicit - Provided
Protein Source
Recombinant, host bacterium Escherichia coli BL21 (DE3) pLysS
Experimental Data (28 measurements)
28 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 (1 hour at 30Β°C), measured by pH-stat (titration assay, using 0.1 M NaOH to keep the pH constant) in aqueous phase | Isopropanol | 50% | 100.0 | β | 116.0 | % | Incubation: ?, Assay: deionized water, 2% gum arabic | Incubation: ?, Assay: ? | Incubation: 30Β°C, Assay: 60Β°C | 2 mg/ml Tricaprylin | Caprylic Acid , glycerol | β | Yes, "Assay preparation by ultrasonic treatment" | Non-incubated control (in %), assay in aqueous phase | Uncertainty about whether assay in aqueous phase or in the presence of organic solvent |
| WT | Stability - Incubation | Activity after incubation (1 hour at 30Β°C), measured by pH-stat (titration assay, using 0.1 M NaOH to keep the pH constant) in aqueous phase | Glycerol | 50% | 100.0 | β | 140.0 | % | Incubation: ?, Assay: deionized water, 2% gum arabic | Incubation: ?, Assay: ? | Incubation: 30Β°C, Assay: 60Β°C | 2 mg/ml Tricaprylin | Caprylic Acid , glycerol | β | Yes, "Assay preparation by ultrasonic treatment" | Non-incubated control (in %), assay in aqueous phase | Uncertainty about whether assay in aqueous phase or in the presence of organic solvent |
| WT | Stability - Incubation | Activity after incubation (1 hour at 30Β°C), measured by pH-stat (titration assay, using 0.1 M NaOH to keep the pH constant) in aqueous phase | Methanol | 50% | 100.0 | β | 106.0 | % | Incubation: ?, Assay: deionized water, 2% gum arabic | Incubation: ?, Assay: ? | Incubation: 30Β°C, Assay: 60Β°C | 2 mg/ml Tricaprylin | Caprylic Acid , glycerol | β | Yes, "Assay preparation by ultrasonic treatment" | Non-incubated control (in %), assay in aqueous phase | Uncertainty about whether assay in aqueous phase or in the presence of organic solvent |
| WT | Stability - Incubation | Activity after incubation (1 hour at 30Β°C), measured by pH-stat (titration assay, using 0.1 M NaOH to keep the pH constant) in aqueous phase | Acetone | 50% | 100.0 | β | 132.0 | % | Incubation: ?, Assay: deionized water, 2% gum arabic | Incubation: ?, Assay: ? | Incubation: 30Β°C, Assay: 60Β°C | 2 mg/ml Tricaprylin | Caprylic Acid , glycerol | β | Yes, "Assay preparation by ultrasonic treatment" | Non-incubated control (in %), assay in aqueous phase | Uncertainty about whether assay in aqueous phase or in the presence of organic solvent |
| WT | Stability - Incubation | Activity after incubation (1 hour at 30Β°C), measured by pH-stat (titration assay, using 0.1 M NaOH to keep the pH constant) in aqueous phase | Chloroform | 50% | 100.0 | β | 89.9 | % | Incubation: ?, Assay: deionized water, 2% gum arabic | Incubation: ?, Assay: ? | Incubation: 30Β°C, Assay: 60Β°C | 2 mg/ml Tricaprylin | Caprylic Acid , glycerol | β | Yes, "Assay preparation by ultrasonic treatment" | Non-incubated control (in %), assay in aqueous phase | Uncertainty about whether assay in aqueous phase or in the presence of organic solvent |
| WT | Stability - Incubation | Activity after incubation (1 hour at 30Β°C), measured by pH-stat (titration assay, using 0.1 M NaOH to keep the pH constant) in aqueous phase | n-Hexane | 50% | 100.0 | β | 120.0 | % | Incubation: ?, Assay: deionized water, 2% gum arabic | Incubation: ?, Assay: ? | Incubation: 30Β°C, Assay: 60Β°C | 2 mg/ml Tricaprylin | Caprylic Acid , glycerol | β | Yes, "Assay preparation by ultrasonic treatment" | Non-incubated control (in %), assay in aqueous phase | Uncertainty about whether assay in aqueous phase or in the presence of organic solvent |
| WT | Stability - Incubation | Activity after incubation (1 hour at 30Β°C), measured by pH-stat (titration assay, using 0.1 M NaOH to keep the pH constant) in aqueous phase | Ethanol | 50% | 100.0 | β | 113.0 | % | Incubation: ?, Assay: deionized water, 2% gum arabic | Incubation: ?, Assay: ? | Incubation: 30Β°C, Assay: 60Β°C | 2 mg/ml Tricaprylin | Caprylic Acid , glycerol | β | Yes, "Assay preparation by ultrasonic treatment" | Non-incubated control (in %), assay in aqueous phase | Uncertainty about whether assay in aqueous phase or in the presence of organic solvent |
| K173A/Q177A | Stability - Incubation | Activity after incubation (1 hour at 30Β°C), measured by pH-stat (titration assay, using 0.1 M NaOH to keep the pH constant) in aqueous phase | Chloroform | 50% | 100.0 | 90.0 | 107.0 | % | Incubation: ?, Assay: deionized water, 2% gum arabic | Incubation: ?, Assay: ? | Incubation: 30Β°C, Assay: 60Β°C | 2 mg/ml Tricaprylin | Caprylic Acid , glycerol | β | Yes, "Assay preparation by ultrasonic treatment" | Non-incubated control (in %), assay in aqueous phase | Uncertainty about whether assay in aqueous phase or in the presence of organic solvent |
| Q177A | Stability - Incubation | Activity after incubation (1 hour at 30Β°C), measured by pH-stat (titration assay, using 0.1 M NaOH to keep the pH constant) in aqueous phase | Chloroform | 50% | 100.0 | 90.0 | 121.0 | % | Incubation: ?, Assay: deionized water, 2% gum arabic | Incubation: ?, Assay: ? | Incubation: 30Β°C, Assay: 60Β°C | 2 mg/ml Tricaprylin | Caprylic Acid , glycerol | β | Yes, "Assay preparation by ultrasonic treatment" | Non-incubated control (in %), assay in aqueous phase | Uncertainty about whether assay in aqueous phase or in the presence of organic solvent |
| Q177A | Stability - Incubation | Activity after incubation (1 hour at 30Β°C), measured by pH-stat (titration assay, using 0.1 M NaOH to keep the pH constant) in aqueous phase | Acetone | 50% | 100.0 | 132.0 | 119.0 | % | Incubation: ?, Assay: deionized water, 2% gum arabic | Incubation: ?, Assay: ? | Incubation: 30Β°C, Assay: 60Β°C | 2 mg/ml Tricaprylin | Caprylic Acid , glycerol | β | Yes, "Assay preparation by ultrasonic treatment" | Non-incubated control (in %), assay in aqueous phase | Uncertainty about whether assay in aqueous phase or in the presence of organic solvent |
| K173A | Stability - Incubation | Activity after incubation (1 hour at 30Β°C), measured by pH-stat (titration assay, using 0.1 M NaOH to keep the pH constant) in aqueous phase | Chloroform | 50% | 100.0 | 90.0 | 121.0 | % | Incubation: ?, Assay: deionized water, 2% gum arabic | Incubation: ?, Assay: ? | Incubation: 30Β°C, Assay: 60Β°C | 2 mg/ml Tricaprylin | Caprylic Acid , glycerol | β | Yes, "Assay preparation by ultrasonic treatment" | Non-incubated control (in %), assay in aqueous phase | Uncertainty about whether assay in aqueous phase or in the presence of organic solvent |
| K173A/Q177A | Stability - Incubation | Activity after incubation (1 hour at 30Β°C), measured by pH-stat (titration assay, using 0.1 M NaOH to keep the pH constant) in aqueous phase | Glycerol | 50% | 100.0 | 140.0 | 104.0 | % | Incubation: ?, Assay: deionized water, 2% gum arabic | Incubation: ?, Assay: ? | Incubation: 30Β°C, Assay: 60Β°C | 2 mg/ml Tricaprylin | Caprylic Acid , glycerol | β | Yes, "Assay preparation by ultrasonic treatment" | Non-incubated control (in %), assay in aqueous phase | Uncertainty about whether assay in aqueous phase or in the presence of organic solvent |
| K173A | Stability - Incubation | Activity after incubation (1 hour at 30Β°C), measured by pH-stat (titration assay, using 0.1 M NaOH to keep the pH constant) in aqueous phase | n-Hexane | 50% | 100.0 | 120.0 | 102.0 | % | Incubation: ?, Assay: deionized water, 2% gum arabic | Incubation: ?, Assay: ? | Incubation: 30Β°C, Assay: 60Β°C | 2 mg/ml Tricaprylin | Caprylic Acid , glycerol | β | Yes, "Assay preparation by ultrasonic treatment" | Non-incubated control (in %), assay in aqueous phase | Uncertainty about whether assay in aqueous phase or in the presence of organic solvent |
| Q177A | Stability - Incubation | Activity after incubation (1 hour at 30Β°C), measured by pH-stat (titration assay, using 0.1 M NaOH to keep the pH constant) in aqueous phase | Glycerol | 50% | 100.0 | 140.0 | 133.0 | % | Incubation: ?, Assay: deionized water, 2% gum arabic | Incubation: ?, Assay: ? | Incubation: 30Β°C, Assay: 60Β°C | 2 mg/ml Tricaprylin | Caprylic Acid , glycerol | β | Yes, "Assay preparation by ultrasonic treatment" | Non-incubated control (in %), assay in aqueous phase | Uncertainty about whether assay in aqueous phase or in the presence of organic solvent |
| Q177A | Stability - Incubation | Activity after incubation (1 hour at 30Β°C), measured by pH-stat (titration assay, using 0.1 M NaOH to keep the pH constant) in aqueous phase | n-Hexane | 50% | 100.0 | 120.0 | 146.0 | % | Incubation: ?, Assay: deionized water, 2% gum arabic | Incubation: ?, Assay: ? | Incubation: 30Β°C, Assay: 60Β°C | 2 mg/ml Tricaprylin | Caprylic Acid , glycerol | β | Yes, "Assay preparation by ultrasonic treatment" | Non-incubated control (in %), assay in aqueous phase | Uncertainty about whether assay in aqueous phase or in the presence of organic solvent |
| K173A | Stability - Incubation | Activity after incubation (1 hour at 30Β°C), measured by pH-stat (titration assay, using 0.1 M NaOH to keep the pH constant) in aqueous phase | Glycerol | 50% | 100.0 | 140.0 | 112.0 | % | Incubation: ?, Assay: deionized water, 2% gum arabic | Incubation: ?, Assay: ? | Incubation: 30Β°C, Assay: 60Β°C | 2 mg/ml Tricaprylin | Caprylic Acid , glycerol | β | Yes, "Assay preparation by ultrasonic treatment" | Non-incubated control (in %), assay in aqueous phase | Uncertainty about whether assay in aqueous phase or in the presence of organic solvent |
| K173A/Q177A | Stability - Incubation | Activity after incubation (1 hour at 30Β°C), measured by pH-stat (titration assay, using 0.1 M NaOH to keep the pH constant) in aqueous phase | n-Hexane | 50% | 100.0 | 120.0 | 108.0 | % | Incubation: ?, Assay: deionized water, 2% gum arabic | Incubation: ?, Assay: ? | Incubation: 30Β°C, Assay: 60Β°C | 2 mg/ml Tricaprylin | Caprylic Acid , glycerol | β | Yes, "Assay preparation by ultrasonic treatment" | Non-incubated control (in %), assay in aqueous phase | Uncertainty about whether assay in aqueous phase or in the presence of organic solvent |
| K173A/Q177A | Stability - Incubation | Activity after incubation (1 hour at 30Β°C), measured by pH-stat (titration assay, using 0.1 M NaOH to keep the pH constant) in aqueous phase | Acetone | 50% | 100.0 | 132.0 | 74.3 | % | Incubation: ?, Assay: deionized water, 2% gum arabic | Incubation: ?, Assay: ? | Incubation: 30Β°C, Assay: 60Β°C | 2 mg/ml Tricaprylin | Caprylic Acid , glycerol | β | Yes, "Assay preparation by ultrasonic treatment" | Non-incubated control (in %), assay in aqueous phase | Uncertainty about whether assay in aqueous phase or in the presence of organic solvent |
| K173A | Stability - Incubation | Activity after incubation (1 hour at 30Β°C), measured by pH-stat (titration assay, using 0.1 M NaOH to keep the pH constant) in aqueous phase | Acetone | 50% | 100.0 | 132.0 | 94.6 | % | Incubation: ?, Assay: deionized water, 2% gum arabic | Incubation: ?, Assay: ? | Incubation: 30Β°C, Assay: 60Β°C | 2 mg/ml Tricaprylin | Caprylic Acid , glycerol | β | Yes, "Assay preparation by ultrasonic treatment" | Non-incubated control (in %), assay in aqueous phase | Uncertainty about whether assay in aqueous phase or in the presence of organic solvent |
| K173A/Q177A | Stability - Incubation | Activity after incubation (1 hour at 30Β°C), measured by pH-stat (titration assay, using 0.1 M NaOH to keep the pH constant) in aqueous phase | Isopropanol | 50% | 100.0 | 116.0 | 104.0 | % | Incubation: ?, Assay: deionized water, 2% gum arabic | Incubation: ?, Assay: ? | Incubation: 30Β°C, Assay: 60Β°C | 2 mg/ml Tricaprylin | Caprylic Acid , glycerol | β | Yes, "Assay preparation by ultrasonic treatment" | Non-incubated control (in %), assay in aqueous phase | Uncertainty about whether assay in aqueous phase or in the presence of organic solvent |
Mutations in this dataset (4) β Parisa Farrokh et al. (2017)
WT
K173A
K173A/Q177A
Q177A
Visualization : Stability β Incubation
Parisa Farrokh et al. (2017)
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.