Lipase from Pseudomonas sp.

Enzyme Description

Species
Pseudomonas sp. β†—
Extremophile
No
EC Number
3.1.1.3 β†—

Sequence

Length: 311 amino acids
MNKNKTLLALCLGSALALSGQAFAATGSGYTATKYPIVLAHGMLGFDSLLGIDYWYGIPSALRRDGAQVYVTEVSQLNTSELRGEELLAQVEEIVAISGKPKVNLIGHSQGGPDIRYVAGVRPDLIASVTSVGAPHKGSDVADLISKVPEGSAGEAVIAGLVNAMGAFINFVSGSSSSAPQNSLGSLESLNSEGAARFNAKFPQGIPTTACGEGAYKVNGVRYYSWSGTSPLTNPLDVSDAMMVAGSLAFSGPNDGLVGRCSSHLGMVIRDNYRMNHLDEVNQFMGLTSLFETDPVSVYRQHANRLKNAGL
Maryam Monsef Shokri et al. (2013) β€” Hydrophobic Substitution of Surface Residues Affects Lipase Stability in Organic Solvents
Molecular Biotechnology  Β· doi:10.1007/s12033-013-9716-y β†—  Β· Stability - Incubation
161 measurements ▼
Database ID
UniProt: C7E3F2 β†—
Sequence Annotation
Explicit - Provided GenBank Accession Number
Protein Source
Recombinant, host bacterium Escherichia coli BL21 (DE3)

Experimental Data (161 measurements)

161 measurements β€” page 4 of 9
Mutation Property Assay Solvent Solvent Volume Aqueous ReferenceWT Reference Measured Value Units Solution pH Temperature Substrate(s) Product(s) Cofactor(s) Shaking Comments
N219D Stability - Incubation Activity after incubation (10 minutes at 60Β°C), measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement) Methanol 25% 65 49 40 % Incubation: 20 mM phosphate buffer, Assay: Phosphate buffer, 10% isopropanol, 0.4% Triton X-100, 1 mg/ml gum arabic Incubation: 7, Assay: 8 Incubation: 60Β°C, Assay: ? 0.6 mM p-Nitrophenyl palmitate p-Nitrophenol , Palmitic Acid β€” Incubation: 150 rpm Water-incubated control (in %), activity measured in aqueous phase | Uncertainty about whether assay was in the presence of organic solvent or in aqueous phase
N219D Stability - Incubation Activity after incubation (20 minutes at 60Β°C), measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement) Methanol 25% 55 β€” 30 % Incubation: 20 mM phosphate buffer, Assay: Phosphate buffer, 10% isopropanol, 0.4% Triton X-100, 1 mg/ml gum arabic Incubation: 7, Assay: 8 Incubation: 60Β°C, Assay: ? 0.6 mM p-Nitrophenyl palmitate p-Nitrophenol , Palmitic Acid β€” Incubation: 150 rpm Water-incubated control (in %), activity measured in aqueous phase | Uncertainty about whether assay was in the presence of organic solvent or in aqueous phase
N219D Stability - Incubation Activity after incubation (30 minutes at 60Β°C), measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement) Methanol 25% 48 29 16 % Incubation: 20 mM phosphate buffer, Assay: Phosphate buffer, 10% isopropanol, 0.4% Triton X-100, 1 mg/ml gum arabic Incubation: 7, Assay: 8 Incubation: 60Β°C, Assay: ? 0.6 mM p-Nitrophenyl palmitate p-Nitrophenol , Palmitic Acid β€” Incubation: 150 rpm Water-incubated control (in %), activity measured in aqueous phase | Uncertainty about whether assay was in the presence of organic solvent or in aqueous phase
N219D Stability - Incubation Activity after incubation (40 minutes at 60Β°C), measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement) Methanol 25% 46 β€” 6 % Incubation: 20 mM phosphate buffer, Assay: Phosphate buffer, 10% isopropanol, 0.4% Triton X-100, 1 mg/ml gum arabic Incubation: 7, Assay: 8 Incubation: 60Β°C, Assay: ? 0.6 mM p-Nitrophenyl palmitate p-Nitrophenol , Palmitic Acid β€” Incubation: 150 rpm Water-incubated control (in %), activity measured in aqueous phase | Uncertainty about whether assay was in the presence of organic solvent or in aqueous phase
S251K Stability - Incubation Activity after incubation (15 minutes at 60Β°C), measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement) Methanol 25% 67 37 64 % Incubation: 20 mM phosphate buffer, Assay: Phosphate buffer, 10% isopropanol, 0.4% Triton X-100, 1 mg/ml gum arabic Incubation: 7, Assay: 8 Incubation: 60Β°C, Assay: ? 0.6 mM p-Nitrophenyl palmitate p-Nitrophenol , Palmitic Acid β€” Incubation: 150 rpm Water-incubated control (in %), activity measured in aqueous phase | Uncertainty about whether assay was in the presence of organic solvent or in aqueous phase
S251K Stability - Incubation Activity after incubation (30 minutes at 60Β°C), measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement) Methanol 25% 60 29 60 % Incubation: 20 mM phosphate buffer, Assay: Phosphate buffer, 10% isopropanol, 0.4% Triton X-100, 1 mg/ml gum arabic Incubation: 7, Assay: 8 Incubation: 60Β°C, Assay: ? 0.6 mM p-Nitrophenyl palmitate p-Nitrophenol , Palmitic Acid β€” Incubation: 150 rpm Water-incubated control (in %), activity measured in aqueous phase | Uncertainty about whether assay was in the presence of organic solvent or in aqueous phase
S251K Stability - Incubation Activity after incubation (40 minutes at 60Β°C), measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement) Methanol 25% 58 β€” 53 % Incubation: 20 mM phosphate buffer, Assay: Phosphate buffer, 10% isopropanol, 0.4% Triton X-100, 1 mg/ml gum arabic Incubation: 7, Assay: 8 Incubation: 60Β°C, Assay: ? 0.6 mM p-Nitrophenyl palmitate p-Nitrophenol , Palmitic Acid β€” Incubation: 150 rpm Water-incubated control (in %), activity measured in aqueous phase | Uncertainty about whether assay was in the presence of organic solvent or in aqueous phase
S251K Stability - Incubation Activity after incubation (50 minutes at 60Β°C), measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement) Methanol 25% 56 β€” 38 % Incubation: 20 mM phosphate buffer, Assay: Phosphate buffer, 10% isopropanol, 0.4% Triton X-100, 1 mg/ml gum arabic Incubation: 7, Assay: 8 Incubation: 60Β°C, Assay: ? 0.6 mM p-Nitrophenyl palmitate p-Nitrophenol , Palmitic Acid β€” Incubation: 150 rpm Water-incubated control (in %), activity measured in aqueous phase | Uncertainty about whether assay was in the presence of organic solvent or in aqueous phase
S251K Stability - Incubation Activity after incubation (60 minutes at 60Β°C), measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement) Methanol 25% 53 25 38 % Incubation: 20 mM phosphate buffer, Assay: Phosphate buffer, 10% isopropanol, 0.4% Triton X-100, 1 mg/ml gum arabic Incubation: 7, Assay: 8 Incubation: 60Β°C, Assay: ? 0.6 mM p-Nitrophenyl palmitate p-Nitrophenol , Palmitic Acid β€” Incubation: 150 rpm Water-incubated control (in %), activity measured in aqueous phase | Uncertainty about whether assay was in the presence of organic solvent or in aqueous phase
S251K Stability - Incubation Activity after incubation (70 minutes at 60Β°C), measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement) Methanol 25% 50 20 32 % Incubation: 20 mM phosphate buffer, Assay: Phosphate buffer, 10% isopropanol, 0.4% Triton X-100, 1 mg/ml gum arabic Incubation: 7, Assay: 8 Incubation: 60Β°C, Assay: ? 0.6 mM p-Nitrophenyl palmitate p-Nitrophenol , Palmitic Acid β€” Incubation: 150 rpm Water-incubated control (in %), activity measured in aqueous phase | Uncertainty about whether assay was in the presence of organic solvent or in aqueous phase
S251L Stability - Incubation Activity after incubation (2.5 minutes at 60Β°C), measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement) Methanol 25% 86 β€” 72 % Incubation: 20 mM phosphate buffer, Assay: Phosphate buffer, 10% isopropanol, 0.4% Triton X-100, 1 mg/ml gum arabic Incubation: 7, Assay: 8 Incubation: 60Β°C, Assay: ? 0.6 mM p-Nitrophenyl palmitate p-Nitrophenol , Palmitic Acid β€” Incubation: 150 rpm Water-incubated control (in %), activity measured in aqueous phase | Uncertainty about whether assay was in the presence of organic solvent or in aqueous phase
S251L Stability - Incubation Activity after incubation (5 minutes at 60Β°C), measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement) Methanol 25% 75 59 60 % Incubation: 20 mM phosphate buffer, Assay: Phosphate buffer, 10% isopropanol, 0.4% Triton X-100, 1 mg/ml gum arabic Incubation: 7, Assay: 8 Incubation: 60Β°C, Assay: ? 0.6 mM p-Nitrophenyl palmitate p-Nitrophenol , Palmitic Acid β€” Incubation: 150 rpm Water-incubated control (in %), activity measured in aqueous phase | Uncertainty about whether assay was in the presence of organic solvent or in aqueous phase
S251L Stability - Incubation Activity after incubation (7.5 minutes at 60Β°C), measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement) Methanol 25% 69 β€” 49 % Incubation: 20 mM phosphate buffer, Assay: Phosphate buffer, 10% isopropanol, 0.4% Triton X-100, 1 mg/ml gum arabic Incubation: 7, Assay: 8 Incubation: 60Β°C, Assay: ? 0.6 mM p-Nitrophenyl palmitate p-Nitrophenol , Palmitic Acid β€” Incubation: 150 rpm Water-incubated control (in %), activity measured in aqueous phase | Uncertainty about whether assay was in the presence of organic solvent or in aqueous phase
S251L Stability - Incubation Activity after incubation (20 minutes at 60Β°C), measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement) Methanol 25% 60 β€” 32 % Incubation: 20 mM phosphate buffer, Assay: Phosphate buffer, 10% isopropanol, 0.4% Triton X-100, 1 mg/ml gum arabic Incubation: 7, Assay: 8 Incubation: 60Β°C, Assay: ? 0.6 mM p-Nitrophenyl palmitate p-Nitrophenol , Palmitic Acid β€” Incubation: 150 rpm Water-incubated control (in %), activity measured in aqueous phase | Uncertainty about whether assay was in the presence of organic solvent or in aqueous phase
S251L Stability - Incubation Activity after incubation (30 minutes at 60Β°C), measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement) Methanol 25% 55 29 18 % Incubation: 20 mM phosphate buffer, Assay: Phosphate buffer, 10% isopropanol, 0.4% Triton X-100, 1 mg/ml gum arabic Incubation: 7, Assay: 8 Incubation: 60Β°C, Assay: ? 0.6 mM p-Nitrophenyl palmitate p-Nitrophenol , Palmitic Acid β€” Incubation: 150 rpm Water-incubated control (in %), activity measured in aqueous phase | Uncertainty about whether assay was in the presence of organic solvent or in aqueous phase
N219L Stability - Incubation Activity after incubation (2 minutes at 60Β°C), measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement) Ethanol 25% 95 β€” 92 % Incubation: 20 mM phosphate buffer, Assay: Phosphate buffer, 10% isopropanol, 0.4% Triton X-100, 1 mg/ml gum arabic Incubation: 7, Assay: 8 Incubation: 60Β°C, Assay: ? 0.6 mM p-Nitrophenyl palmitate p-Nitrophenol , Palmitic Acid β€” Incubation: 150 rpm Water-incubated control (in %), activity measured in aqueous phase | Uncertainty about whether assay was in the presence of organic solvent or in aqueous phase
N219L Stability - Incubation Activity after incubation (15 minutes at 60Β°C), measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement) Ethanol 25% 86 β€” 50 % Incubation: 20 mM phosphate buffer, Assay: Phosphate buffer, 10% isopropanol, 0.4% Triton X-100, 1 mg/ml gum arabic Incubation: 7, Assay: 8 Incubation: 60Β°C, Assay: ? 0.6 mM p-Nitrophenyl palmitate p-Nitrophenol , Palmitic Acid β€” Incubation: 150 rpm Water-incubated control (in %), activity measured in aqueous phase | Uncertainty about whether assay was in the presence of organic solvent or in aqueous phase
N219L Stability - Incubation Activity after incubation (30 minutes at 60Β°C), measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement) Ethanol 25% 83 β€” 36 % Incubation: 20 mM phosphate buffer, Assay: Phosphate buffer, 10% isopropanol, 0.4% Triton X-100, 1 mg/ml gum arabic Incubation: 7, Assay: 8 Incubation: 60Β°C, Assay: ? 0.6 mM p-Nitrophenyl palmitate p-Nitrophenol , Palmitic Acid β€” Incubation: 150 rpm Water-incubated control (in %), activity measured in aqueous phase | Uncertainty about whether assay was in the presence of organic solvent or in aqueous phase
N219L Stability - Incubation Activity after incubation (40 minutes at 60Β°C), measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement) Ethanol 25% 82 β€” 28 % Incubation: 20 mM phosphate buffer, Assay: Phosphate buffer, 10% isopropanol, 0.4% Triton X-100, 1 mg/ml gum arabic Incubation: 7, Assay: 8 Incubation: 60Β°C, Assay: ? 0.6 mM p-Nitrophenyl palmitate p-Nitrophenol , Palmitic Acid β€” Incubation: 150 rpm Water-incubated control (in %), activity measured in aqueous phase | Uncertainty about whether assay was in the presence of organic solvent or in aqueous phase
N219I Stability - Incubation Activity after incubation (10 minutes at 60Β°C), measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement) Ethanol 25% 88 14 50 % Incubation: 20 mM phosphate buffer, Assay: Phosphate buffer, 10% isopropanol, 0.4% Triton X-100, 1 mg/ml gum arabic Incubation: 7, Assay: 8 Incubation: 60Β°C, Assay: ? 0.6 mM p-Nitrophenyl palmitate p-Nitrophenol , Palmitic Acid β€” Incubation: 150 rpm Water-incubated control (in %), activity measured in aqueous phase | Uncertainty about whether assay was in the presence of organic solvent or in aqueous phase
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Mutations in this dataset (9)

WT N219A N219D N219I N219L N219P N219R S251K S251L

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.

Structure

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