Cholesterol esterase (gene cheS) from Burkholderia cepacia ST-200

Enzyme Description

Species

Extremophile

EC Number

3.1.1.13 ↗

Sequence

Length: 364 amino acids

MARSMRSRVVAGAVACAMSVAPFAGTTALMTLATTHAAMAATAPADNYAATRYPIILVHGLTGTDKYAGVLEYWYGIQEDLQQHGATVYVANLSGFQSDDGPNGRGEQLLAYVKTVLAATGAAKVNLVGHSQGGLTSRYVAAVAPDLVASVTTIGTPHRGSEFADFVQSVLAYDPTGLSSTVIAAFVNVFGILTSSSHNTNQDALASLKTLTTSQAATYNQNYPSAGLGAPGSCQTGAPTETVGGNTHLLYSWAGTAIQPTLSLFGVTGAQDTSTIPLVDPANALDPSTLALFGTGTVMINRGSGQNDGLVSKCSALYGKVLSTSYKWNHIDEINQLLGVRGAYAEDPVAVIRTHANRLQLAGV

Yasuhiko Takeda et al. (2006) — Purification, characterization, and molecular cloning of organic-solvent-tolerant cholesterol esterase from cyclohexane-tolerant Burkholderia cepacia strain ST-200

Extremophiles · doi:10.1007/s00792-005-0494-8 ↗ · Activity - Classical Stability - Incubation

30 measurements ▼

Database ID

UniProt: Q75NT4 ↗

Sequence Annotation

Explicit - Provided

Protein Source

Recombinant, host bacterium Escherichia coli

Experimental Data (30 measurements)

30 measurements — page 1 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	Methanol	10%	1.0	6.8	relative units	180 mM potassium phosphate buffer	7	37°C	0.8 mM p-Nitrophenyl caprylate	Caprylic Acid , p-Nitrophenol	—	—	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	Acetone	50%	1.0	0.21	relative units	180 mM potassium phosphate buffer	7	37°C	0.8 mM p-Nitrophenyl caprylate	Caprylic Acid , p-Nitrophenol	—	—	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	Acetone	20%	1.0	3.8	relative units	180 mM potassium phosphate buffer	7	37°C	0.8 mM p-Nitrophenyl caprylate	Caprylic Acid , p-Nitrophenol	—	—	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	Acetone	10%	1.0	4.6	relative units	180 mM potassium phosphate buffer	7	37°C	0.8 mM p-Nitrophenyl caprylate	Caprylic Acid , p-Nitrophenol	—	—	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	Acetone	5%	1.0	3.5	relative units	180 mM potassium phosphate buffer	7	37°C	0.8 mM p-Nitrophenyl caprylate	Caprylic Acid , p-Nitrophenol	—	—	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	Isopropanol	50%	1.0	0.23	relative units	180 mM potassium phosphate buffer	7	37°C	0.8 mM p-Nitrophenyl caprylate	Caprylic Acid , p-Nitrophenol	—	—	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	Isopropanol	20%	1.0	5.1	relative units	180 mM potassium phosphate buffer	7	37°C	0.8 mM p-Nitrophenyl caprylate	Caprylic Acid , p-Nitrophenol	—	—	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	Isopropanol	10%	1.0	5.7	relative units	180 mM potassium phosphate buffer	7	37°C	0.8 mM p-Nitrophenyl caprylate	Caprylic Acid , p-Nitrophenol	—	—	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	Isopropanol	5%	1.0	5.8	relative units	180 mM potassium phosphate buffer	7	37°C	0.8 mM p-Nitrophenyl caprylate	Caprylic Acid , p-Nitrophenol	—	—	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	Ethanol	50%	1.0	0.43	relative units	180 mM potassium phosphate buffer	7	37°C	0.8 mM p-Nitrophenyl caprylate	Caprylic Acid , p-Nitrophenol	—	—	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	Ethanol	20%	1.0	4.6	relative units	180 mM potassium phosphate buffer	7	37°C	0.8 mM p-Nitrophenyl caprylate	Caprylic Acid , p-Nitrophenol	—	—	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	Ethanol	10%	1.0	5.8	relative units	180 mM potassium phosphate buffer	7	37°C	0.8 mM p-Nitrophenyl caprylate	Caprylic Acid , p-Nitrophenol	—	—	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	Ethanol	5%	1.0	5.4	relative units	180 mM potassium phosphate buffer	7	37°C	0.8 mM p-Nitrophenyl caprylate	Caprylic Acid , p-Nitrophenol	—	—	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	Methanol	50%	1.0	2.8	relative units	180 mM potassium phosphate buffer	7	37°C	0.8 mM p-Nitrophenyl caprylate	Caprylic Acid , p-Nitrophenol	—	—	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	Methanol	20%	1.0	7.2	relative units	180 mM potassium phosphate buffer	7	37°C	0.8 mM p-Nitrophenyl caprylate	Caprylic Acid , p-Nitrophenol	—	—	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	Methanol	5%	1.0	6.5	relative units	180 mM potassium phosphate buffer	7	37°C	0.8 mM p-Nitrophenyl caprylate	Caprylic Acid , p-Nitrophenol	—	—	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	Dimethylformamide (DMF)	50%	1.0	3.1	relative units	180 mM potassium phosphate buffer	7	37°C	0.8 mM p-Nitrophenyl caprylate	Caprylic Acid , p-Nitrophenol	—	—	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	Dimethylformamide (DMF)	20%	1.0	6.0	relative units	180 mM potassium phosphate buffer	7	37°C	0.8 mM p-Nitrophenyl caprylate	Caprylic Acid , p-Nitrophenol	—	—	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	Dimethylformamide (DMF)	10%	1.0	5.4	relative units	180 mM potassium phosphate buffer	7	37°C	0.8 mM p-Nitrophenyl caprylate	Caprylic Acid , p-Nitrophenol	—	—	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	Dimethylformamide (DMF)	5%	1.0	4.9	relative units	180 mM potassium phosphate buffer	7	37°C	0.8 mM p-Nitrophenyl caprylate	Caprylic Acid , p-Nitrophenol	—	—	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

Methanol

10%

1.0

6.8

relative units

180 mM potassium phosphate buffer

37°C

0.8 mM p-Nitrophenyl caprylate

Caprylic Acid, p-Nitrophenol

—

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

Acetone

50%

1.0

0.21

relative units

180 mM potassium phosphate buffer

37°C

0.8 mM p-Nitrophenyl caprylate

Caprylic Acid, p-Nitrophenol

—

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

Acetone

20%

1.0

3.8

relative units

180 mM potassium phosphate buffer

37°C

0.8 mM p-Nitrophenyl caprylate

Caprylic Acid, p-Nitrophenol

—

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

Acetone

10%

1.0

4.6

relative units

180 mM potassium phosphate buffer

37°C

0.8 mM p-Nitrophenyl caprylate

Caprylic Acid, p-Nitrophenol

—

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

Acetone

1.0

3.5

relative units

180 mM potassium phosphate buffer

37°C

0.8 mM p-Nitrophenyl caprylate

Caprylic Acid, p-Nitrophenol

—

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

Isopropanol

50%

1.0

0.23

relative units

180 mM potassium phosphate buffer

37°C

0.8 mM p-Nitrophenyl caprylate

Caprylic Acid, p-Nitrophenol

—

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

Isopropanol

20%

1.0

5.1

relative units

180 mM potassium phosphate buffer

37°C

0.8 mM p-Nitrophenyl caprylate

Caprylic Acid, p-Nitrophenol

—

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

Isopropanol

10%

1.0

5.7

relative units

180 mM potassium phosphate buffer

37°C

0.8 mM p-Nitrophenyl caprylate

Caprylic Acid, p-Nitrophenol

—

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

Isopropanol

1.0

5.8

relative units

180 mM potassium phosphate buffer

37°C

0.8 mM p-Nitrophenyl caprylate

Caprylic Acid, p-Nitrophenol

—

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

Ethanol

50%

1.0

0.43

relative units

180 mM potassium phosphate buffer

37°C

0.8 mM p-Nitrophenyl caprylate

Caprylic Acid, p-Nitrophenol

—

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

Ethanol

20%

1.0

4.6

relative units

180 mM potassium phosphate buffer

37°C

0.8 mM p-Nitrophenyl caprylate

Caprylic Acid, p-Nitrophenol

—

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

Ethanol

10%

1.0

5.8

relative units

180 mM potassium phosphate buffer

37°C

0.8 mM p-Nitrophenyl caprylate

Caprylic Acid, p-Nitrophenol

—

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

Ethanol

1.0

5.4

relative units

180 mM potassium phosphate buffer

37°C

0.8 mM p-Nitrophenyl caprylate

Caprylic Acid, p-Nitrophenol

—

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

Methanol

50%

1.0

2.8

relative units

180 mM potassium phosphate buffer

37°C

0.8 mM p-Nitrophenyl caprylate

Caprylic Acid, p-Nitrophenol

—

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

Methanol

20%

1.0

7.2

relative units

180 mM potassium phosphate buffer

37°C

0.8 mM p-Nitrophenyl caprylate

Caprylic Acid, p-Nitrophenol

—

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

Methanol

1.0

6.5

relative units

180 mM potassium phosphate buffer

37°C

0.8 mM p-Nitrophenyl caprylate

Caprylic Acid, p-Nitrophenol

—

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

Dimethylformamide (DMF)

50%

1.0

3.1

relative units

180 mM potassium phosphate buffer

37°C

0.8 mM p-Nitrophenyl caprylate

Caprylic Acid, p-Nitrophenol

—

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

Dimethylformamide (DMF)

20%

1.0

6.0

relative units

180 mM potassium phosphate buffer

37°C

0.8 mM p-Nitrophenyl caprylate

Caprylic Acid, p-Nitrophenol

—

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

Dimethylformamide (DMF)

10%

1.0

5.4

relative units

180 mM potassium phosphate buffer

37°C

0.8 mM p-Nitrophenyl caprylate

Caprylic Acid, p-Nitrophenol

—

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

Dimethylformamide (DMF)

1.0

4.9

relative units

180 mM potassium phosphate buffer

37°C

0.8 mM p-Nitrophenyl caprylate

Caprylic Acid, p-Nitrophenol

—

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)

50%

1.0

4.3

relative units

180 mM potassium phosphate buffer

37°C

0.8 mM p-Nitrophenyl caprylate

Caprylic Acid, p-Nitrophenol

—

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

37°C

0.8 mM p-Nitrophenyl caprylate

Caprylic Acid, p-Nitrophenol

—

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

37°C

0.8 mM p-Nitrophenyl caprylate

Caprylic Acid, p-Nitrophenol

—

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)

1.0

4.8

relative units

180 mM potassium phosphate buffer

37°C

0.8 mM p-Nitrophenyl caprylate

Caprylic Acid, p-Nitrophenol

—

Classical aqueous control (in relative units)

Stability - Incubation

Activity after incubation (24 hours at 30°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: 30°C, Assay: 37°C

20 mM Cholesterol linoleate

Cholesterol, linoleic acid

—

Non-incubated control (in relative units), assay in aqueous phase | Uncertainty about whether the assay really was in aqueous phase

Stability - Incubation

Isopropanol

33%

1.0

1.1

relative units

Incubation: 10 mM Tris-HCl, Assay: 0.11 M potassium phosphate

Incubation: 8, Assay: 7

Incubation: 30°C, Assay: 37°C

20 mM Cholesterol linoleate

Cholesterol, linoleic acid

—

Non-incubated control (in relative units), assay in aqueous phase | Uncertainty about whether the assay really was in aqueous phase

Stability - Incubation

Acetone

33%

1.0

0.53

relative units

Incubation: 10 mM Tris-HCl, Assay: 0.11 M potassium phosphate

Incubation: 8, Assay: 7

Incubation: 30°C, Assay: 37°C

20 mM Cholesterol linoleate

Cholesterol, linoleic acid

—

Non-incubated control (in relative units), assay in aqueous phase | Uncertainty about whether the assay really was in aqueous phase

Stability - Incubation

Ethanol

33%

1.0

0.98

relative units

Incubation: 10 mM Tris-HCl, Assay: 0.11 M potassium phosphate

Incubation: 8, Assay: 7

Incubation: 30°C, Assay: 37°C

20 mM Cholesterol linoleate

Cholesterol, linoleic acid

—

Non-incubated control (in relative units), assay in aqueous phase | Uncertainty about whether the assay really was in aqueous phase

Stability - Incubation

Methanol

33%

1.0

0.58

relative units

Incubation: 10 mM Tris-HCl, Assay: 0.11 M potassium phosphate

Incubation: 8, Assay: 7

Incubation: 30°C, Assay: 37°C

20 mM Cholesterol linoleate

Cholesterol, linoleic acid

—

Non-incubated control (in relative units), assay in aqueous phase | Uncertainty about whether the assay really was in aqueous phase

Stability - Incubation

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: 30°C, Assay: 37°C

20 mM Cholesterol linoleate

Cholesterol, linoleic acid

—

Non-incubated control (in relative units), assay in aqueous phase | Uncertainty about whether the assay really was in aqueous phase

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Visualization : Activity — Classical

One bar per measurement. Colour = solvent, shade = solvent volume.

Visualization : Stability — Incubation

One bar per measurement. Colour = solvent, shade = solvent volume.

Structure

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Cholesterol esterase (gene cheS) from Burkholderia cepacia ST-200

Enzyme Description

Sequence

Experimental Data (30 measurements)

Visualization : Activity — Classical

Visualization : Stability — Incubation

Structure

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