Cholesterol oxidase from Burkholderia cepacia ST-200
Enzyme Description
Sequence
ADAPPPGFPADIPLYKQAFQNWSGEIAVQDVWTAAPRSADDVVAAVNWARANGYRIRPRGYMHNWSPLTLDPGAGAANVVLLDTTKSLTAVSVDTSARPARVTAQTGISLESLLATLEQYGLGVIAAPAPGDITLGGALAIDAHGTAVPAVGETLQPGHTYGSLSNLVVALTAVVYDPARQQYVLRRFERSDPEIGAFLAHIGRAFVVEVTLTAGPNQRLRCQSYVDIPASELFAPAGTSGRTITSFLDRAGRVEAIWFPFTSSPWLKVWTPTPSKPFLSRAVTQPYNYPFSDSISQSISDLVKRIVIGGEGALTPLFGQTQLAITAAGLALTLSGDIWGWSRTVLQYIRPTTLRVTANGYAVLARRADVQRVISEFVQFYQNRVDTYKARGEYPMNGPVEIRITGLDKPADAGAGAAVPSLSALKPRPDRPEWDVAVWFDILTLPGTPSADRFYREIEQWMLANYTGSYATLRPEWSKGWGYTDTAAWQDDTMLTTTIPNLQREGQPASSTWDTARATLERYDPHRIFRSPLLDRLMP
Noriyuki Doukyu et al. (1998)
β Purification of Extracellular Cholesterol Oxidase with High Activity in the Presence of Organic Solvents from Pseudomonas sp. Strain ST-200
Applied and Environmental Microbiology
Β· doi:10.1128/AEM.64.5.1929-1932.1998 β
Β· Activity + Stability - Product Formation
▼
Database ID
UniProt: Q93RE1 β
Sequence Annotation
Inferred - from protein name (first 43 residues from UniProt sequence absent from the mature protein)
Protein Source
Purified, bacterium Pseudomonas sp. strain ST-200
Experimental Data (6 measurements)
6 measurements
| Property | Assay | Solvent | Solvent Volume | Aqueous Reference | Measured Value | Units | Solution | pH | Temperature | Substrate(s) | Product(s) | Cofactor(s) | Shaking | Comments |
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| Activity + Stability - Product Formation | Product formation after incubation (3 hours at 30Β°C) in the presence of organic solvent measured by HPLC | Chloroform | 33.3% | 1.0 | 0.2 | relative units | 50 mM sodium potassium phosphate, 15 mM sodium cholate | Incubation: 7, Assay: 7 | 30Β°C | 2.5 mg/ml in the organic solvent Cholesterol | Cholest-4-en-3-one , H2O2 | β | Yes, incubation "with shaking" | Classical aqueous control (in relative units) | Organism named Pseudomonas strain ST-200, but reclassified as Burkolderia cepacia strain ST-200 (see 10.1007/s002530100753) |
| Activity + Stability - Product Formation | Product formation after incubation (3 hours at 30Β°C) in the presence of organic solvent measured by HPLC | Benzene | 33.3% | 1.0 | 3.0 | relative units | 50 mM sodium potassium phosphate, 15 mM sodium cholate | Incubation: 7, Assay: 7 | 30Β°C | 2.5 mg/ml in the organic solvent Cholesterol | Cholest-4-en-3-one , H2O2 | β | Yes, incubation "with shaking" | Classical aqueous control (in relative units) | Organism named Pseudomonas strain ST-200, but reclassified as Burkolderia cepacia strain ST-200 (see 10.1007/s002530100753) |
| Activity + Stability - Product Formation | Product formation after incubation (3 hours at 30Β°C) in the presence of organic solvent measured by HPLC | Toluene | 33.3% | 1.0 | 3.5 | relative units | 50 mM sodium potassium phosphate, 15 mM sodium cholate | Incubation: 7, Assay: 7 | 30Β°C | 2.5 mg/ml in the organic solvent Cholesterol | Cholest-4-en-3-one , H2O2 | β | Yes, incubation "with shaking" | Classical aqueous control (in relative units) | Organism named Pseudomonas strain ST-200, but reclassified as Burkolderia cepacia strain ST-200 (see 10.1007/s002530100753) |
| Activity + Stability - Product Formation | Product formation after incubation (3 hours at 30Β°C) in the presence of organic solvent measured by HPLC | p-Xylene | 33.3% | 1.0 | 3.4 | relative units | 50 mM sodium potassium phosphate, 15 mM sodium cholate | Incubation: 7, Assay: 7 | 30Β°C | 2.5 mg/ml in the organic solvent Cholesterol | Cholest-4-en-3-one , H2O2 | β | Yes, incubation "with shaking" | Classical aqueous control (in relative units) | Organism named Pseudomonas strain ST-200, but reclassified as Burkolderia cepacia strain ST-200 (see 10.1007/s002530100753) |
| Activity + Stability - Product Formation | Product formation after incubation (3 hours at 30Β°C) in the presence of organic solvent measured by HPLC | Propylbenzene | 33.3% | 1.0 | 3.2 | relative units | 50 mM sodium potassium phosphate, 15 mM sodium cholate | Incubation: 7, Assay: 7 | 30Β°C | 2.5 mg/ml in the organic solvent Cholesterol | Cholest-4-en-3-one , H2O2 | β | Yes, incubation "with shaking" | Classical aqueous control (in relative units) | Organism named Pseudomonas strain ST-200, but reclassified as Burkolderia cepacia strain ST-200 (see 10.1007/s002530100753) |
| Activity + Stability - Product Formation | Product formation after incubation (3 hours at 30Β°C) in the presence of organic solvent measured by HPLC | Diphenylmethane | 33.3% | 1.0 | 3.1 | relative units | 50 mM sodium potassium phosphate, 15 mM sodium cholate | Incubation: 7, Assay: 7 | 30Β°C | 2.5 mg/ml in the organic solvent Cholesterol | Cholest-4-en-3-one , H2O2 | β | Yes, incubation "with shaking" | Classical aqueous control (in relative units) | Organism named Pseudomonas strain ST-200, but reclassified as Burkolderia cepacia strain ST-200 (see 10.1007/s002530100753) |
Visualization : Activity + Stability β Product Formation
Noriyuki Doukyu et al. (1998)
One bar per measurement. Colour = solvent, shade = solvent volume.
Doukyu N. et al. (2001)
β Cloning, sequence analysis and expression of a gene encoding an organic solvent- and detergent-tolerant cholesterol oxidase of Burkholderia cepacia strain ST-200
Applied Microbiology and Biotechnology
Β· doi:10.1007/s002530100753 β
Β· Stability - Incubation
▼
Database ID
UniProt: Q93RE1 β
Sequence Annotation
Explicit - Provided (first 43 residues from UniProt sequence absent from the mature protein)
Protein Source
Recombinant, host bacterium Escherichia coli DH5alpha
Experimental Data (10 measurements)
10 measurements
| Property | Assay | Solvent | Solvent Volume | Aqueous Reference | Measured Value | Units | Solution | pH | Temperature | Substrate(s) | Product(s) | Cofactor(s) | Shaking | Comments |
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| Stability - Incubation | Activity after incubation (12 hours at 37Β°C), measured by absorbance spectrophotometry (H2O2 absorbance measurement) in aqueous phase | Acetone | 33.3% | 100 | 22 | % | Incubation: 50 mM sodium potassium phosphate, 15 mM sodium cholate, Assay: 50 mM sodium potassium phosphate, 15 mM sodium cholate | Incubation: 7, Assay: 7 | Incubation: 37Β°C, Assay: 30Β°C | 2.5 mg/ml in the organic solvent Cholesterol | Cholest-4-en-3-one , H2O2 | β | Yes, incubation "with shaking" | Non-incubated control (in %), assay in aqueous phase | Clear about assay in aqueous phase |
| Stability - Incubation | Activity after incubation (12 hours at 37Β°C), measured by absorbance spectrophotometry (H2O2 absorbance measurement) in aqueous phase | Isopropanol | 33.3% | 100 | 75 | % | Incubation: 50 mM sodium potassium phosphate, 15 mM sodium cholate, Assay: 50 mM sodium potassium phosphate, 15 mM sodium cholate | Incubation: 7, Assay: 7 | Incubation: 37Β°C, Assay: 30Β°C | 2.5 mg/ml in the organic solvent Cholesterol | Cholest-4-en-3-one , H2O2 | β | Yes, incubation "with shaking" | Non-incubated control (in %), assay in aqueous phase | Clear about assay in aqueous phase |
| Stability - Incubation | Activity after incubation (12 hours at 37Β°C), measured by absorbance spectrophotometry (H2O2 absorbance measurement) in aqueous phase | Ethyl Acetate | 33.3% | 100 | 102 | % | Incubation: 50 mM sodium potassium phosphate, 15 mM sodium cholate, Assay: 50 mM sodium potassium phosphate, 15 mM sodium cholate | Incubation: 7, Assay: 7 | Incubation: 37Β°C, Assay: 30Β°C | 2.5 mg/ml in the organic solvent Cholesterol | Cholest-4-en-3-one , H2O2 | β | Yes, incubation "with shaking" | Non-incubated control (in %), assay in aqueous phase | Clear about assay in aqueous phase |
| Stability - Incubation | Activity after incubation (12 hours at 37Β°C), measured by absorbance spectrophotometry (H2O2 absorbance measurement) in aqueous phase | 1-Butanol | 33.3% | 100 | 77 | % | Incubation: 50 mM sodium potassium phosphate, 15 mM sodium cholate, Assay: 50 mM sodium potassium phosphate, 15 mM sodium cholate | Incubation: 7, Assay: 7 | Incubation: 37Β°C, Assay: 30Β°C | 2.5 mg/ml in the organic solvent Cholesterol | Cholest-4-en-3-one , H2O2 | β | Yes, incubation "with shaking" | Non-incubated control (in %), assay in aqueous phase | Clear about assay in aqueous phase |
| Stability - Incubation | Activity after incubation (12 hours at 37Β°C), measured by absorbance spectrophotometry (H2O2 absorbance measurement) in aqueous phase | Chloroform | 33.3% | 100 | 97 | % | Incubation: 50 mM sodium potassium phosphate, 15 mM sodium cholate, Assay: 50 mM sodium potassium phosphate, 15 mM sodium cholate | Incubation: 7, Assay: 7 | Incubation: 37Β°C, Assay: 30Β°C | 2.5 mg/ml in the organic solvent Cholesterol | Cholest-4-en-3-one , H2O2 | β | Yes, incubation "with shaking" | Non-incubated control (in %), assay in aqueous phase | Clear about assay in aqueous phase |
| Stability - Incubation | Activity after incubation (12 hours at 37Β°C), measured by absorbance spectrophotometry (H2O2 absorbance measurement) in aqueous phase | Benzene | 33.3% | 100 | 99 | % | Incubation: 50 mM sodium potassium phosphate, 15 mM sodium cholate, Assay: 50 mM sodium potassium phosphate, 15 mM sodium cholate | Incubation: 7, Assay: 7 | Incubation: 37Β°C, Assay: 30Β°C | 2.5 mg/ml in the organic solvent Cholesterol | Cholest-4-en-3-one , H2O2 | β | Yes, incubation "with shaking" | Non-incubated control (in %), assay in aqueous phase | Clear about assay in aqueous phase |
| Stability - Incubation | Activity after incubation (12 hours at 37Β°C), measured by absorbance spectrophotometry (H2O2 absorbance measurement) in aqueous phase | Toluene | 33.3% | 100 | 98 | % | Incubation: 50 mM sodium potassium phosphate, 15 mM sodium cholate, Assay: 50 mM sodium potassium phosphate, 15 mM sodium cholate | Incubation: 7, Assay: 7 | Incubation: 37Β°C, Assay: 30Β°C | 2.5 mg/ml in the organic solvent Cholesterol | Cholest-4-en-3-one , H2O2 | β | Yes, incubation "with shaking" | Non-incubated control (in %), assay in aqueous phase | Clear about assay in aqueous phase |
| Stability - Incubation | Activity after incubation (12 hours at 37Β°C), measured by absorbance spectrophotometry (H2O2 absorbance measurement) in aqueous phase | p-Xylene | 33.3% | 100 | 99 | % | Incubation: 50 mM sodium potassium phosphate, 15 mM sodium cholate, Assay: 50 mM sodium potassium phosphate, 15 mM sodium cholate | Incubation: 7, Assay: 7 | Incubation: 37Β°C, Assay: 30Β°C | 2.5 mg/ml in the organic solvent Cholesterol | Cholest-4-en-3-one , H2O2 | β | Yes, incubation "with shaking" | Non-incubated control (in %), assay in aqueous phase | Clear about assay in aqueous phase |
| Stability - Incubation | Activity after incubation (12 hours at 37Β°C), measured by absorbance spectrophotometry (H2O2 absorbance measurement) in aqueous phase | Diphenylmethane | 33.3% | 100 | 98 | % | Incubation: 50 mM sodium potassium phosphate, 15 mM sodium cholate, Assay: 50 mM sodium potassium phosphate, 15 mM sodium cholate | Incubation: 7, Assay: 7 | Incubation: 37Β°C, Assay: 30Β°C | 2.5 mg/ml in the organic solvent Cholesterol | Cholest-4-en-3-one , H2O2 | β | Yes, incubation "with shaking" | Non-incubated control (in %), assay in aqueous phase | Clear about assay in aqueous phase |
| Stability - Incubation | Activity after incubation (12 hours at 37Β°C), measured by absorbance spectrophotometry (H2O2 absorbance measurement) in aqueous phase | Cyclooctane | 33.3% | 100 | 99 | % | Incubation: 50 mM sodium potassium phosphate, 15 mM sodium cholate, Assay: 50 mM sodium potassium phosphate, 15 mM sodium cholate | Incubation: 7, Assay: 7 | Incubation: 37Β°C, Assay: 30Β°C | 2.5 mg/ml in the organic solvent Cholesterol | Cholest-4-en-3-one , H2O2 | β | Yes, incubation "with shaking" | Non-incubated control (in %), assay in aqueous phase | Clear about assay in aqueous phase |
Visualization : Stability β Incubation
Doukyu N. et al. (2001)
One bar per measurement. Colour = solvent, shade = solvent volume.