Cholesterol oxidase from Chromobacterium sp. DS-1

Enzyme Description

Species
Chromobacterium sp. DS-1 β†—
Extremophile
No
EC Number
1.1.3.6 β†—

Sequence

Length: 540 amino acids
TCSQPNNFPAEIPLYKQSFKNWAGDIKVDDVWTCAPRSADEVVKVANWAKDNGYKVRARGMMHNWSPLTLAAGVSCPAVVLLDTTRYLTAMSIDASGPVAKVTAQAGITMEALLTGLEKAGLGVTAAPAPGDLTLGGVLAINGHGTAIPAKGERRLAGASYGSISNLVLSLTAVVYDKASGAYALRKFARNDPQIAPLLAHVGRSLIVEATLQAAPNQRLRCQSWFNIPYGEMFAAAGSGGRTFASYLDSAGRVEAIWFPFTSNPWLKVWTVTPNKPLFSRQTDKPFNYPFSDNLPDEVTDLANKILSLGDGKLTPAFGKAQFAAASAGLVATASWDLWGWSKNLLLYVKPTTLRVTANGYAVLTRRENVQRVLNEFVTFYQARVQAYQQQGRYPMNGPVEIRVTGLDDPSEAALSGGVAPALSAIRPRPDHPEWNVAVWLDILTLPGTPYANQFYREIEQWIEANFNGSYAAVRPEWSKGWGYTDQAAWADSAMLQTTIPNAFRAGQPAAANWDAAKAALAAYDPYRLFSSPLLDSLGL
Noriyuki Doukyu et al. (2009) β€” Cloning, sequence analysis, and expression of a gene encoding Chromobacterium sp. DS-1 cholesterol oxidase
Applied Microbiology and Biotechnology  Β· doi:10.1007/s00253-008-1775-9 β†—  Β· Stability - Incubation
12 measurements ▼
Database ID
UniProt: B5MGF8 β†—
Sequence Annotation
Explicit - Provided (first 44 from UniPrort sequence removed in mature protein)
Protein Source
Recombinant, host bacterium Escherichia coli BL21 (DE3) pLysS

Experimental Data (12 measurements)

12 measurements
Property Assay Solvent Solvent Volume Aqueous Reference Measured Value Units Extraction method Data type Solution pH Temperature Substrate(s) Product(s) Cofactor(s) Shaking Comments
Stability - Incubation Activity after incubation (24 hours at 37Β°C) measured by absorbance spectrophotometry (colorimetric assay, red quinoneimine formed by H2O2 reaction with 4-aminoantipyrine and phenol in the presence of HRP absorbance measurement, 500 nm) in aqueous phase Dimethyl Sulfoxide (DMSO) 33 % v/v 100 99 % Direct Continuous Incubation: 50 mM sodium phosphate buffer, Assay: 50 mM sodium potassium phosphate buffer, 64 mM sodium cholate, 0.34% Triton X-100, 1.4 mM 4-aminoantipyrine, 21 mM phenol, 0.89 mM cholesterol, and 5 U/ml horseradish peroxidase Incubation: 7, Assay: 7 Incubation: 37Β°C, Assay: 30Β°C Cholesterol, Oβ‚‚ Cholest-4-en-3-one , H2​O2 FAD Yes, "mixture incubated with shaking" Non-incubated control (in %), residual activity | Assay parameters taken from cited article (10.1128/AEM.64.5.1929-1932.1998)
Stability - Incubation Activity after incubation (24 hours at 37Β°C) measured by absorbance spectrophotometry (colorimetric assay, red quinoneimine formed by H2O2 reaction with 4-aminoantipyrine and phenol in the presence of HRP absorbance measurement, 500 nm) in aqueous phase Methanol 33 % v/v 100 108 % Direct Continuous Incubation: 50 mM sodium phosphate buffer, Assay: 50 mM sodium potassium phosphate buffer, 64 mM sodium cholate, 0.34% Triton X-100, 1.4 mM 4-aminoantipyrine, 21 mM phenol, 0.89 mM cholesterol, and 5 U/ml horseradish peroxidase Incubation: 7, Assay: 7 Incubation: 37Β°C, Assay: 30Β°C Cholesterol, Oβ‚‚ Cholest-4-en-3-one , H2​O2 FAD Yes, "mixture incubated with shaking" Non-incubated control (in %), residual activity | Assay parameters taken from cited article (10.1128/AEM.64.5.1929-1932.1998)
Stability - Incubation Activity after incubation (24 hours at 37Β°C) measured by absorbance spectrophotometry (colorimetric assay, red quinoneimine formed by H2O2 reaction with 4-aminoantipyrine and phenol in the presence of HRP absorbance measurement, 500 nm) in aqueous phase Ethanol 33 % v/v 100 111 % Direct Continuous Incubation: 50 mM sodium phosphate buffer, Assay: 50 mM sodium potassium phosphate buffer, 64 mM sodium cholate, 0.34% Triton X-100, 1.4 mM 4-aminoantipyrine, 21 mM phenol, 0.89 mM cholesterol, and 5 U/ml horseradish peroxidase Incubation: 7, Assay: 7 Incubation: 37Β°C, Assay: 30Β°C Cholesterol, Oβ‚‚ Cholest-4-en-3-one , H2​O2 FAD Yes, "mixture incubated with shaking" Non-incubated control (in %), residual activity | Assay parameters taken from cited article (10.1128/AEM.64.5.1929-1932.1998)
Stability - Incubation Activity after incubation (24 hours at 37Β°C) measured by absorbance spectrophotometry (colorimetric assay, red quinoneimine formed by H2O2 reaction with 4-aminoantipyrine and phenol in the presence of HRP absorbance measurement, 500 nm) in aqueous phase Acetone 33 % v/v 100 15 % Direct Continuous Incubation: 50 mM sodium phosphate buffer, Assay: 50 mM sodium potassium phosphate buffer, 64 mM sodium cholate, 0.34% Triton X-100, 1.4 mM 4-aminoantipyrine, 21 mM phenol, 0.89 mM cholesterol, and 5 U/ml horseradish peroxidase Incubation: 7, Assay: 7 Incubation: 37Β°C, Assay: 30Β°C Cholesterol, Oβ‚‚ Cholest-4-en-3-one , H2​O2 FAD Yes, "mixture incubated with shaking" Non-incubated control (in %), residual activity | Assay parameters taken from cited article (10.1128/AEM.64.5.1929-1932.1998)
Stability - Incubation Activity after incubation (24 hours at 37Β°C) measured by absorbance spectrophotometry (colorimetric assay, red quinoneimine formed by H2O2 reaction with 4-aminoantipyrine and phenol in the presence of HRP absorbance measurement, 500 nm) in aqueous phase Isopropanol 33 % v/v 100 116 % Direct Continuous Incubation: 50 mM sodium phosphate buffer, Assay: 50 mM sodium potassium phosphate buffer, 64 mM sodium cholate, 0.34% Triton X-100, 1.4 mM 4-aminoantipyrine, 21 mM phenol, 0.89 mM cholesterol, and 5 U/ml horseradish peroxidase Incubation: 7, Assay: 7 Incubation: 37Β°C, Assay: 30Β°C Cholesterol, Oβ‚‚ Cholest-4-en-3-one , H2​O2 FAD Yes, "mixture incubated with shaking" Non-incubated control (in %), residual activity | Assay parameters taken from cited article (10.1128/AEM.64.5.1929-1932.1998)
Stability - Incubation Activity after incubation (24 hours at 37Β°C) measured by absorbance spectrophotometry (colorimetric assay, red quinoneimine formed by H2O2 reaction with 4-aminoantipyrine and phenol in the presence of HRP absorbance measurement, 500 nm) in aqueous phase Ethyl Acetate 33 % v/v 100 98 % Direct Continuous Incubation: 50 mM sodium phosphate buffer, Assay: 50 mM sodium potassium phosphate buffer, 64 mM sodium cholate, 0.34% Triton X-100, 1.4 mM 4-aminoantipyrine, 21 mM phenol, 0.89 mM cholesterol, and 5 U/ml horseradish peroxidase Incubation: 7, Assay: 7 Incubation: 37Β°C, Assay: 30Β°C Cholesterol, Oβ‚‚ Cholest-4-en-3-one , H2​O2 FAD Yes, "mixture incubated with shaking" Non-incubated control (in %), residual activity | Assay parameters taken from cited article (10.1128/AEM.64.5.1929-1932.1998)
Stability - Incubation Activity after incubation (24 hours at 37Β°C) measured by absorbance spectrophotometry (colorimetric assay, red quinoneimine formed by H2O2 reaction with 4-aminoantipyrine and phenol in the presence of HRP absorbance measurement, 500 nm) in aqueous phase 1-Butanol 33 % v/v 100 99 % Direct Continuous Incubation: 50 mM sodium phosphate buffer, Assay: 50 mM sodium potassium phosphate buffer, 64 mM sodium cholate, 0.34% Triton X-100, 1.4 mM 4-aminoantipyrine, 21 mM phenol, 0.89 mM cholesterol, and 5 U/ml horseradish peroxidase Incubation: 7, Assay: 7 Incubation: 37Β°C, Assay: 30Β°C Cholesterol, Oβ‚‚ Cholest-4-en-3-one , H2​O2 FAD Yes, "mixture incubated with shaking" Non-incubated control (in %), residual activity | Assay parameters taken from cited article (10.1128/AEM.64.5.1929-1932.1998)
Stability - Incubation Activity after incubation (24 hours at 37Β°C) measured by absorbance spectrophotometry (colorimetric assay, red quinoneimine formed by H2O2 reaction with 4-aminoantipyrine and phenol in the presence of HRP absorbance measurement, 500 nm) in aqueous phase Chloroform 33 % v/v 100 115 % Direct Continuous Incubation: 50 mM sodium phosphate buffer, Assay: 50 mM sodium potassium phosphate buffer, 64 mM sodium cholate, 0.34% Triton X-100, 1.4 mM 4-aminoantipyrine, 21 mM phenol, 0.89 mM cholesterol, and 5 U/ml horseradish peroxidase Incubation: 7, Assay: 7 Incubation: 37Β°C, Assay: 30Β°C Cholesterol, Oβ‚‚ Cholest-4-en-3-one , H2​O2 FAD Yes, "mixture incubated with shaking" Non-incubated control (in %), residual activity | Assay parameters taken from cited article (10.1128/AEM.64.5.1929-1932.1998)
Stability - Incubation Activity after incubation (24 hours at 37Β°C) measured by absorbance spectrophotometry (colorimetric assay, red quinoneimine formed by H2O2 reaction with 4-aminoantipyrine and phenol in the presence of HRP absorbance measurement, 500 nm) in aqueous phase Benzene 33 % v/v 100 112 % Direct Continuous Incubation: 50 mM sodium phosphate buffer, Assay: 50 mM sodium potassium phosphate buffer, 64 mM sodium cholate, 0.34% Triton X-100, 1.4 mM 4-aminoantipyrine, 21 mM phenol, 0.89 mM cholesterol, and 5 U/ml horseradish peroxidase Incubation: 7, Assay: 7 Incubation: 37Β°C, Assay: 30Β°C Cholesterol, Oβ‚‚ Cholest-4-en-3-one , H2​O2 FAD Yes, "mixture incubated with shaking" Non-incubated control (in %), residual activity | Assay parameters taken from cited article (10.1128/AEM.64.5.1929-1932.1998)
Stability - Incubation Activity after incubation (24 hours at 37Β°C) measured by absorbance spectrophotometry (colorimetric assay, red quinoneimine formed by H2O2 reaction with 4-aminoantipyrine and phenol in the presence of HRP absorbance measurement, 500 nm) in aqueous phase Toluene 33 % v/v 100 110 % Direct Continuous Incubation: 50 mM sodium phosphate buffer, Assay: 50 mM sodium potassium phosphate buffer, 64 mM sodium cholate, 0.34% Triton X-100, 1.4 mM 4-aminoantipyrine, 21 mM phenol, 0.89 mM cholesterol, and 5 U/ml horseradish peroxidase Incubation: 7, Assay: 7 Incubation: 37Β°C, Assay: 30Β°C Cholesterol, Oβ‚‚ Cholest-4-en-3-one , H2​O2 FAD Yes, "mixture incubated with shaking" Non-incubated control (in %), residual activity | Assay parameters taken from cited article (10.1128/AEM.64.5.1929-1932.1998)
Stability - Incubation Activity after incubation (24 hours at 37Β°C) measured by absorbance spectrophotometry (colorimetric assay, red quinoneimine formed by H2O2 reaction with 4-aminoantipyrine and phenol in the presence of HRP absorbance measurement, 500 nm) in aqueous phase p-Xylene 33 % v/v 100 102 % Direct Continuous Incubation: 50 mM sodium phosphate buffer, Assay: 50 mM sodium potassium phosphate buffer, 64 mM sodium cholate, 0.34% Triton X-100, 1.4 mM 4-aminoantipyrine, 21 mM phenol, 0.89 mM cholesterol, and 5 U/ml horseradish peroxidase Incubation: 7, Assay: 7 Incubation: 37Β°C, Assay: 30Β°C Cholesterol, Oβ‚‚ Cholest-4-en-3-one , H2​O2 FAD Yes, "mixture incubated with shaking" Non-incubated control (in %), residual activity | Assay parameters taken from cited article (10.1128/AEM.64.5.1929-1932.1998)
Stability - Incubation Activity after incubation (24 hours at 37Β°C) measured by absorbance spectrophotometry (colorimetric assay, red quinoneimine formed by H2O2 reaction with 4-aminoantipyrine and phenol in the presence of HRP absorbance measurement, 500 nm) in aqueous phase Cyclooctane 33 % v/v 100 102 % Direct Continuous Incubation: 50 mM sodium phosphate buffer, Assay: 50 mM sodium potassium phosphate buffer, 64 mM sodium cholate, 0.34% Triton X-100, 1.4 mM 4-aminoantipyrine, 21 mM phenol, 0.89 mM cholesterol, and 5 U/ml horseradish peroxidase Incubation: 7, Assay: 7 Incubation: 37Β°C, Assay: 30Β°C Cholesterol, Oβ‚‚ Cholest-4-en-3-one , H2​O2 FAD Yes, "mixture incubated with shaking" Non-incubated control (in %), residual activity | Assay parameters taken from cited article (10.1128/AEM.64.5.1929-1932.1998)

Visualization : Stability β€” Incubation

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

Structure

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