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Literature summary for 1.14.15.22 extracted from
- Three-step hydroxylation of vitamin D3 by a genetically engineered CYP105A1: enzymes and catalysis (2010), FEBS J., 277, 3999-4009.
Application
| Application | Comment | Organism |
|---|---|---|
| synthesis | expression of mutant R73V/R84A in Streptomyces lividans TK23 cells under the control of the tipA promoter leads to synthesis of 25-hydroxyvitamin D3 and large amounts of 1alpha,25-dihydroxyvitamin D3. In addition, polar metabolites 1alpha,25(R),26-trihydroxyvitamin D3 and 1alpha,25(R),26-trihydroxyvitamin D3 are observed at a ratio of 5:1 | Streptomyces griseolus |
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| expression in Streptomyces lividans | Streptomyces griseolus |
Crystallization (Commentary)
| Crystallization (Comment) | Organism |
|---|---|
| structure of the mutant R73V/R84A with 1alpha,25-dihydroxyvitamin D3, a hydrogen-bond network including the 1alpha-hydroxyl group and several water molecules play an important role in the substrate-binding for 26-hydroxylation | Streptomyces griseolus |
Protein Variants
| Protein Variants | Comment | Organism |
|---|---|---|
| R73V/R84A | highly active mutant, kinetic and crystallization data | Streptomyces griseolus |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.0012 | - |
1alpha,25-dihydroxyvitamin D3 | 26-hydroxylation, mutant R73V/R84A, pH not specified in the publication, temperature not specified in the publication | Streptomyces griseolus |  |
| 0.0022 | - |
25-hydroxyvitamin D3 | 1alpha-hydroxylation, mutant R73V/R84A, pH not specified in the publication, temperature not specified in the publication | Streptomyces griseolus | |
| 0.0035 | - |
vitamin D3 | 25-hydroxylation, mutant R73V/R84A, pH not specified in the publication, temperature not specified in the publication | Streptomyces griseolus | |
| 0.0065 | - |
1alpha-hydroxyvitamin D3 | 25-hydroxylation, mutant R73V/R84A, pH not specified in the publication, temperature not specified in the publication | Streptomyces griseolus | |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Streptomyces griseolus | P18326 | - |
- |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| 1alpha,25-dihydroxyvitamin D3 + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2 | - |
Streptomyces griseolus | 1alpha,25(R),26-trihydroxyvitamin D3 + 1alpha,25(S),26-trihydroxyvitamin D3 + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O | reaction of mutant R73V/R84A, 25R- and 25S-enantiomers are found in a ratio of 5:1 | ? | |
| 1alpha-hydroxyvitamin D3 + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2 | - |
Streptomyces griseolus | 1alpha,25-dihydroxyvitamin D3 + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O | - |
? | |
| 25-hydroxyvitamin D3 + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2 | - |
Streptomyces griseolus | 1alpha,25-dihydroxyvitamin D3 + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O | - |
? | |
| vitamin D3 + O2 + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ | - |
Streptomyces griseolus | 25-hydroxyvitamin D3 + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O | - |
? | |
Turnover Number [1/s]
| Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.0009 | - |
1alpha,25-dihydroxyvitamin D3 | 26-hydroxylation, mutant R73V/R84A, pH not specified in the publication, temperature not specified in the publication | Streptomyces griseolus | |
| 0.0023 | - |
25-hydroxyvitamin D3 | 1alpha-hydroxylation, mutant R73V/R84A, pH not specified in the publication, temperature not specified in the publication | Streptomyces griseolus | |
| 0.0023 | - |
vitamin D3 | 25-hydroxylation, mutant R73V/R84A, pH not specified in the publication, temperature not specified in the publication | Streptomyces griseolus | |
| 0.035 | - |
1alpha-hydroxyvitamin D3 | 25-hydroxylation, mutant R73V/R84A, pH not specified in the publication, temperature not specified in the publication | Streptomyces griseolus | |
kcat/KM [mM/s]
| kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.67 | - |
vitamin D3 | 25-hydroxylation, mutant R73V/R84A, pH not specified in the publication, temperature not specified in the publication | Streptomyces griseolus | |
| 0.72 | - |
1alpha,25-dihydroxyvitamin D3 | 26-hydroxylation, mutant R73V/R84A, pH not specified in the publication, temperature not specified in the publication | Streptomyces griseolus | |
| 1.03 | - |
25-hydroxyvitamin D3 | 1alpha-hydroxylation, mutant R73V/R84A, pH not specified in the publication, temperature not specified in the publication | Streptomyces griseolus | |
| 5.5 | - |
1alpha-hydroxyvitamin D3 | 25-hydroxylation, mutant R73V/R84A, pH not specified in the publication, temperature not specified in the publication | Streptomyces griseolus | |
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