Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli BL21(DE3)pLysS cells | Streptomyces sp. |
Crystallization (Comment) | Organism |
---|---|
mutant enzymes N485D and N485L, hanging drop vapor diffusion method, using 9-11% (w/v) polyethylene glycol 8000, 75 mM MnSO4 and 100 mM sodium cacodylate pH 5.2 | Streptomyces sp. |
Protein Variants | Comment | Organism |
---|---|---|
N485D | the kcat of N485D is diminished about 650times compared with that of wild type, while the apparent Km value is minimally affected. For the N485D mutant, pH 5.1results in a 20fold increase in the rate of oxidation compared with that at pH 7.0 | Streptomyces sp. |
N485L | the kcat of N485D is diminished about 1110times compared with that of wild type, while the apparent Km value is minimally affected | Streptomyces sp. |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0027 | - |
cholesterol | apparent value, wild type enzyme, pH 7.0, temperature not specified in the publication | Streptomyces sp. | |
0.0046 | - |
cholesterol | apparent value, mutant enzyme N485L, pH 7.0, temperature not specified in the publication | Streptomyces sp. | |
0.0062 | - |
cholesterol | apparent value, mutant enzyme N485L, pH 5.1, temperature not specified in the publication | Streptomyces sp. | |
0.0063 | - |
cholesterol | apparent value, mutant enzyme N485D, pH 5.1, temperature not specified in the publication | Streptomyces sp. | |
0.0066 | - |
cholesterol | apparent value, mutant enzyme N485D, pH 7.0, temperature not specified in the publication | Streptomyces sp. | |
0.0067 | - |
cholesterol | apparent value, wild type enzyme, pH 5.1, temperature not specified in the publication | Streptomyces sp. |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Streptomyces sp. | P12676 | - |
- |
Purification (Comment) | Organism |
---|---|
butyl-Sepharose column chromatography and DEAE-cellulose column chromatography | Streptomyces sp. |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
cholesterol + O2 | - |
Streptomyces sp. | cholest-4-en-3-one + H2O2 | - |
? |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.035 | - |
cholesterol | apparent value, mutant enzyme N485L, pH 5.1, temperature not specified in the publication | Streptomyces sp. | |
0.044 | - |
cholesterol | apparent value, mutant enzyme N485L, pH 7.0, temperature not specified in the publication | Streptomyces sp. | |
0.073 | - |
cholesterol | apparent value, mutant enzyme N485D, pH 7.0, temperature not specified in the publication | Streptomyces sp. | |
1.4 | - |
cholesterol | apparent value, mutant enzyme N485D, pH 5.1, temperature not specified in the publication | Streptomyces sp. | |
46 | - |
cholesterol | apparent value, wild type enzyme, pH 5.1, temperature not specified in the publication | Streptomyces sp. | |
47 | - |
cholesterol | apparent value, wild type enzyme, pH 7.0, temperature not specified in the publication | Streptomyces sp. |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
FAD | - |
Streptomyces sp. |