Cholesterol esterase (gene cheS) from Burkholderia cepacia ST-200
Enzyme Description
Sequence
MARSMRSRVVAGAVACAMSVAPFAGTTALMTLATTHAAMAATAPADNYAATRYPIILVHGLTGTDKYAGVLEYWYGIQEDLQQHGATVYVANLSGFQSDDGPNGRGEQLLAYVKTVLAATGAAKVNLVGHSQGGLTSRYVAAVAPDLVASVTTIGTPHRGSEFADFVQSVLAYDPTGLSSTVIAAFVNVFGILTSSSHNTNQDALASLKTLTTSQAATYNQNYPSAGLGAPGSCQTGAPTETVGGNTHLLYSWAGTAIQPTLSLFGVTGAQDTSTIPLVDPANALDPSTLALFGTGTVMINRGSGQNDGLVSKCSALYGKVLSTSYKWNHIDEINQLLGVRGAYAEDPVAVIRTHANRLQLAGV
Yasuhiko Takeda et al. (2006)
β Purification, characterization, and molecular cloning of organic-solvent-tolerant cholesterol esterase from cyclohexane-tolerant Burkholderia cepacia strain ST-200
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Database ID
UniProt: Q75NT4 β
Sequence Annotation
Explicit - Provided
Protein Source
Recombinant, host bacterium Escherichia coli
Experimental Data (30 measurements)
30 measurements β page 2 of 2
| Property | Assay | Solvent | Solvent Volume | Aqueous Reference | Measured Value | Units | Solution | pH | Temperature | Substrate(s) | Product(s) | Cofactor(s) | Shaking | Comments |
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| Activity - Classical | Activity measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 410 nm) in the presence of organic solvent | Dimethyl Sulfoxide (DMSO) | 50% | 1.0 | 4.3 | relative units | 180 mM potassium phosphate buffer | 7 | 37Β°C | 0.8 mM p-Nitrophenyl caprylate | Caprylic Acid , p-Nitrophenol | β | β | Classical aqueous control (in relative units) |
| Activity - Classical | Activity measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 410 nm) in the presence of organic solvent | Dimethyl Sulfoxide (DMSO) | 20% | 1.0 | 5.6 | relative units | 180 mM potassium phosphate buffer | 7 | 37Β°C | 0.8 mM p-Nitrophenyl caprylate | Caprylic Acid , p-Nitrophenol | β | β | Classical aqueous control (in relative units) |
| Activity - Classical | Activity measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 410 nm) in the presence of organic solvent | Dimethyl Sulfoxide (DMSO) | 10% | 1.0 | 5.0 | relative units | 180 mM potassium phosphate buffer | 7 | 37Β°C | 0.8 mM p-Nitrophenyl caprylate | Caprylic Acid , p-Nitrophenol | β | β | Classical aqueous control (in relative units) |
| Activity - Classical | Activity measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 410 nm) in the presence of organic solvent | Dimethyl Sulfoxide (DMSO) | 5% | 1.0 | 4.8 | relative units | 180 mM potassium phosphate buffer | 7 | 37Β°C | 0.8 mM p-Nitrophenyl caprylate | Caprylic Acid , p-Nitrophenol | β | β | Classical aqueous control (in relative units) |
| Stability - Incubation | Activity after incubation (24 hours at 30Β°C), measured by absorbance spectrophotometry (colorimetric Allain's method, cholesterol and O2 cholesterol oxidase-catalyzed reaction produces H2O2, and H2O2 and 4-Aminoantipyrine reaction product absorbance measurement, 500 nm) in aqueous phase | Dimethyl Sulfoxide (DMSO) | 33% | 1.0 | 1.1 | relative units | Incubation: 10 mM Tris-HCl, Assay: 0.11 M potassium phosphate | Incubation: 8, Assay: 7 | Incubation: 30Β°C, Assay: 37Β°C | 20 mM Cholesterol linoleate | Cholesterol , linoleic acid | β | β | Non-incubated control (in relative units), assay in aqueous phase | Uncertainty about whether the assay really was in aqueous phase |
| Stability - Incubation | Activity after incubation (24 hours at 30Β°C), measured by absorbance spectrophotometry (colorimetric Allain's method, cholesterol and O2 cholesterol oxidase-catalyzed reaction produces H2O2, and H2O2 and 4-Aminoantipyrine reaction product absorbance measurement, 500 nm) in aqueous phase | Isopropanol | 33% | 1.0 | 1.1 | relative units | Incubation: 10 mM Tris-HCl, Assay: 0.11 M potassium phosphate | Incubation: 8, Assay: 7 | Incubation: 30Β°C, Assay: 37Β°C | 20 mM Cholesterol linoleate | Cholesterol , linoleic acid | β | β | Non-incubated control (in relative units), assay in aqueous phase | Uncertainty about whether the assay really was in aqueous phase |
| Stability - Incubation | Activity after incubation (24 hours at 30Β°C), measured by absorbance spectrophotometry (colorimetric Allain's method, cholesterol and O2 cholesterol oxidase-catalyzed reaction produces H2O2, and H2O2 and 4-Aminoantipyrine reaction product absorbance measurement, 500 nm) in aqueous phase | Acetone | 33% | 1.0 | 0.53 | relative units | Incubation: 10 mM Tris-HCl, Assay: 0.11 M potassium phosphate | Incubation: 8, Assay: 7 | Incubation: 30Β°C, Assay: 37Β°C | 20 mM Cholesterol linoleate | Cholesterol , linoleic acid | β | β | Non-incubated control (in relative units), assay in aqueous phase | Uncertainty about whether the assay really was in aqueous phase |
| Stability - Incubation | Activity after incubation (24 hours at 30Β°C), measured by absorbance spectrophotometry (colorimetric Allain's method, cholesterol and O2 cholesterol oxidase-catalyzed reaction produces H2O2, and H2O2 and 4-Aminoantipyrine reaction product absorbance measurement, 500 nm) in aqueous phase | Ethanol | 33% | 1.0 | 0.98 | relative units | Incubation: 10 mM Tris-HCl, Assay: 0.11 M potassium phosphate | Incubation: 8, Assay: 7 | Incubation: 30Β°C, Assay: 37Β°C | 20 mM Cholesterol linoleate | Cholesterol , linoleic acid | β | β | Non-incubated control (in relative units), assay in aqueous phase | Uncertainty about whether the assay really was in aqueous phase |
| Stability - Incubation | Activity after incubation (24 hours at 30Β°C), measured by absorbance spectrophotometry (colorimetric Allain's method, cholesterol and O2 cholesterol oxidase-catalyzed reaction produces H2O2, and H2O2 and 4-Aminoantipyrine reaction product absorbance measurement, 500 nm) in aqueous phase | Methanol | 33% | 1.0 | 0.58 | relative units | Incubation: 10 mM Tris-HCl, Assay: 0.11 M potassium phosphate | Incubation: 8, Assay: 7 | Incubation: 30Β°C, Assay: 37Β°C | 20 mM Cholesterol linoleate | Cholesterol , linoleic acid | β | β | Non-incubated control (in relative units), assay in aqueous phase | Uncertainty about whether the assay really was in aqueous phase |
| Stability - Incubation | Activity after incubation (24 hours at 30Β°C), measured by absorbance spectrophotometry (colorimetric Allain's method, cholesterol and O2 cholesterol oxidase-catalyzed reaction produces H2O2, and H2O2 and 4-Aminoantipyrine reaction product absorbance measurement, 500 nm) in aqueous phase | Dimethylformamide (DMF) | 33% | 1.0 | 0.98 | relative units | Incubation: 10 mM Tris-HCl, Assay: 0.11 M potassium phosphate | Incubation: 8, Assay: 7 | Incubation: 30Β°C, Assay: 37Β°C | 20 mM Cholesterol linoleate | Cholesterol , linoleic acid | β | β | Non-incubated control (in relative units), assay in aqueous phase | Uncertainty about whether the assay really was in aqueous phase |
Visualization : Activity β Classical
One bar per measurement. Colour = solvent, shade = solvent volume.
Visualization : Stability β Incubation
One bar per measurement. Colour = solvent, shade = solvent volume.