Lipase (gene lipB) from Serratia marcescens ECU1010

Enzyme Description

Species
Serratia marcescens ECU1010 β†—
Extremophile
No
EC Number
3.1.1.3 β†—

Sequence

Length: 614 amino acids
MGIFSYKDLDENASKALFSDALAISTYAYHNIGNGFDEGYHQTGFGLGLPLTLVTALIGSTQSQGGLPGLPWNPDSEQAAQEAVNNAGWSVISAAQLGYAGKTDARGTYYGETAGYTTAQAEVLGKYDSEGNLTAIGISFRGTSGPRESLIGDTIGDVINDLLAGFGPKGYADGYTLKAFGNLLGDAAKFAQAHGLSGEDVVVSGHSLGGLAVNSMAAQSDASWSGFYAQSNYVAFASPTQYETGGKVLNIGYENDPVFRALDGTTLTGTSLGVHDAPHTSATNNIVNFNDHYASDAWNLLPFSILNIPTWLSHLPFFYQDGLMRVLNSEFYSLTDKDSTIIVSNLSNITRGNTWVEDLNRNAETHSGPTFIIGSDGNDLIKGGKGNDYLEGRDGDDIFRDAGGYNLIAGGKGHNIFDTQQALKNTEVAYDGNTLYLRDAKGGITLADDISTLRSKETSWLIFNKEVDHQVTAAGLKSDSGLKAYAAAATGGDGDDVLQARSHDAWLFGNAGNDTLIGHAGGNLTFVGGSGDDVLKGVGNGNTFLFSGDFGRDQLYGFNASDKLVFIGTEGASGNIRDYATQQNDDLVLAFGHSQVTLIGVSLDHFSTDQVVLA
Su-Xia Li et al. (2014) β€” Essential Role of Gly33 in a Novel Organic Solvent-Tolerant Lipase from Serratia marcescens ECU1010 as Determined by Site-Directed Mutagenesis
Applied Biochemistry and Biotechnology  Β· doi:10.1007/s12010-013-0690-4 β†—  Β· Stability - Incubation
72 measurements ▼
Database ID
UniProt: D7SFB5 β†—
Sequence Annotation
Explicit - Provided
Protein Source
Recombinant, host bacterium Escherichia coli BL21 (DE3)

Experimental Data (72 measurements)

72 measurements β€” page 4 of 4
Mutation Property Assay Solvent Solvent Volume Aqueous ReferenceWT Reference Measured Value Units Solution pH Temperature Substrate(s) Product(s) Cofactor(s) Shaking Comments
G33D Stability - Incubation Activity after incubation (24 hours at 30Β°C), measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in aqueous phase Acetone 99% 100.0 33.7 23.5 % Incubation: 100 mM potassium phosphate buffer, Assay: 100 mM potassium phosphate buffer Incubation: 8, Assay: 7.5 Incubation: 30Β°C, Assay: 30Β°C ? mM p-Nitrophenyl butyrate Butyric Acid , p-Nitrophenol β€” Incubation: 160 rpm Non-incubated control (in %), activity measured in aqueous phase | Uncertainty about whether assay was in the presence of organic solvent or in aqueous phase
G33D Stability - Incubation Activity after incubation (24 hours at 30Β°C), measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in aqueous phase Diisopropyl Ether 10% 100.0 168.3 101.5 % Incubation: 100 mM potassium phosphate buffer, Assay: 100 mM potassium phosphate buffer Incubation: 8, Assay: 7.5 Incubation: 30Β°C, Assay: 30Β°C ? mM p-Nitrophenyl butyrate Butyric Acid , p-Nitrophenol β€” Incubation: 160 rpm Non-incubated control (in %), activity measured in aqueous phase | Uncertainty about whether assay was in the presence of organic solvent or in aqueous phase
G33D Stability - Incubation Activity after incubation (24 hours at 30Β°C), measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in aqueous phase Diisopropyl Ether 50% 100.0 316.8 129.6 % Incubation: 100 mM potassium phosphate buffer, Assay: 100 mM potassium phosphate buffer Incubation: 8, Assay: 7.5 Incubation: 30Β°C, Assay: 30Β°C ? mM p-Nitrophenyl butyrate Butyric Acid , p-Nitrophenol β€” Incubation: 160 rpm Non-incubated control (in %), activity measured in aqueous phase | Uncertainty about whether assay was in the presence of organic solvent or in aqueous phase
G33D Stability - Incubation Activity after incubation (24 hours at 30Β°C), measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in aqueous phase Diisopropyl Ether 99% 100.0 297.0 114.3 % Incubation: 100 mM potassium phosphate buffer, Assay: 100 mM potassium phosphate buffer Incubation: 8, Assay: 7.5 Incubation: 30Β°C, Assay: 30Β°C ? mM p-Nitrophenyl butyrate Butyric Acid , p-Nitrophenol β€” Incubation: 160 rpm Non-incubated control (in %), activity measured in aqueous phase | Uncertainty about whether assay was in the presence of organic solvent or in aqueous phase
A187V Stability - Incubation Activity after incubation (24 hours at 30Β°C), measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in aqueous phase Dimethyl Sulfoxide (DMSO) 10% 100.0 57.6 47.5 % Incubation: 100 mM potassium phosphate buffer, Assay: 100 mM potassium phosphate buffer Incubation: 8, Assay: 7.5 Incubation: 30Β°C, Assay: 30Β°C ? mM p-Nitrophenyl butyrate Butyric Acid , p-Nitrophenol β€” Incubation: 160 rpm Non-incubated control (in %), activity measured in aqueous phase | Uncertainty about whether assay was in the presence of organic solvent or in aqueous phase
A187V Stability - Incubation Activity after incubation (24 hours at 30Β°C), measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in aqueous phase Dimethyl Sulfoxide (DMSO) 50% 100.0 8.1 6.4 % Incubation: 100 mM potassium phosphate buffer, Assay: 100 mM potassium phosphate buffer Incubation: 8, Assay: 7.5 Incubation: 30Β°C, Assay: 30Β°C ? mM p-Nitrophenyl butyrate Butyric Acid , p-Nitrophenol β€” Incubation: 160 rpm Non-incubated control (in %), activity measured in aqueous phase | Uncertainty about whether assay was in the presence of organic solvent or in aqueous phase
A187V Stability - Incubation Activity after incubation (24 hours at 30Β°C), measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in aqueous phase Dimethyl Sulfoxide (DMSO) 99% 100.0 1.6 2.0 % Incubation: 100 mM potassium phosphate buffer, Assay: 100 mM potassium phosphate buffer Incubation: 8, Assay: 7.5 Incubation: 30Β°C, Assay: 30Β°C ? mM p-Nitrophenyl butyrate Butyric Acid , p-Nitrophenol β€” Incubation: 160 rpm Non-incubated control (in %), activity measured in aqueous phase | Uncertainty about whether assay was in the presence of organic solvent or in aqueous phase
A187V Stability - Incubation Activity after incubation (24 hours at 30Β°C), measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in aqueous phase Methanol 10% 100.0 72.3 83.3 % Incubation: 100 mM potassium phosphate buffer, Assay: 100 mM potassium phosphate buffer Incubation: 8, Assay: 7.5 Incubation: 30Β°C, Assay: 30Β°C ? mM p-Nitrophenyl butyrate Butyric Acid , p-Nitrophenol β€” Incubation: 160 rpm Non-incubated control (in %), activity measured in aqueous phase | Uncertainty about whether assay was in the presence of organic solvent or in aqueous phase
A187V Stability - Incubation Activity after incubation (24 hours at 30Β°C), measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in aqueous phase Methanol 50% 100.0 1.8 2.0 % Incubation: 100 mM potassium phosphate buffer, Assay: 100 mM potassium phosphate buffer Incubation: 8, Assay: 7.5 Incubation: 30Β°C, Assay: 30Β°C ? mM p-Nitrophenyl butyrate Butyric Acid , p-Nitrophenol β€” Incubation: 160 rpm Non-incubated control (in %), activity measured in aqueous phase | Uncertainty about whether assay was in the presence of organic solvent or in aqueous phase
A187V Stability - Incubation Activity after incubation (24 hours at 30Β°C), measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in aqueous phase Methanol 99% 100.0 4.6 5.2 % Incubation: 100 mM potassium phosphate buffer, Assay: 100 mM potassium phosphate buffer Incubation: 8, Assay: 7.5 Incubation: 30Β°C, Assay: 30Β°C ? mM p-Nitrophenyl butyrate Butyric Acid , p-Nitrophenol β€” Incubation: 160 rpm Non-incubated control (in %), activity measured in aqueous phase | Uncertainty about whether assay was in the presence of organic solvent or in aqueous phase
A187V Stability - Incubation Activity after incubation (24 hours at 30Β°C), measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in aqueous phase Isopropanol 10% 100.0 51.5 66.6 % Incubation: 100 mM potassium phosphate buffer, Assay: 100 mM potassium phosphate buffer Incubation: 8, Assay: 7.5 Incubation: 30Β°C, Assay: 30Β°C ? mM p-Nitrophenyl butyrate Butyric Acid , p-Nitrophenol β€” Incubation: 160 rpm Non-incubated control (in %), activity measured in aqueous phase | Uncertainty about whether assay was in the presence of organic solvent or in aqueous phase
A187V Stability - Incubation Activity after incubation (24 hours at 30Β°C), measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in aqueous phase Isopropanol 50% 100.0 2.0 4.7 % Incubation: 100 mM potassium phosphate buffer, Assay: 100 mM potassium phosphate buffer Incubation: 8, Assay: 7.5 Incubation: 30Β°C, Assay: 30Β°C ? mM p-Nitrophenyl butyrate Butyric Acid , p-Nitrophenol β€” Incubation: 160 rpm Non-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 (4)

WT G33D A187V T270A

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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