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Literature summary extracted from
- Purification, characterization, and directed evolution study of a vitamin D3 hydroxylase from Pseudonocardia autotrophica (2009), Biochem. Biophys. Res. Commun., 385, 170-175.
Activating Compound
| EC Number | Activating Compound | Comment | Organism | Structure |
|---|---|---|---|---|
| 1.14.14.24 | KCl | or NaCl, required. Optimum concentration 100 mM | Pseudonocardia autotrophica |  |
| 1.14.14.24 | NaCl | or KCl, required. Optimum concentration 100 mM | Pseudonocardia autotrophica | |
| 1.14.14.24 | partially methylated beta-cyclodextrin | activity increases approximately 10fold when partially methylated beta-cyclodextrin is present in the reaction at a final concentration of 0.05% | Pseudonocardia autotrophica |
Cloned(Commentary)
| EC Number | Cloned (Comment) | Organism |
|---|---|---|
| 1.14.14.24 | expression in Escherichia coli | Pseudonocardia autotrophica |
Protein Variants
| EC Number | Protein Variants | Comment | Organism |
|---|---|---|---|
| 1.14.14.24 | E216A | 2- to 3fold increase in hydroxylase activity | Pseudonocardia autotrophica |
| 1.14.14.24 | E384R | 2- to 3fold increase in hydroxylase activity | Pseudonocardia autotrophica |
| 1.14.14.24 | T70R | 2- to 3fold increase in hydroxylase activity | Pseudonocardia autotrophica |
| 1.14.14.24 | T70R/V156L/E216M/E384R | 21fold increase in hydroxylase activity | Pseudonocardia autotrophica |
| 1.14.14.24 | T70R/V156S/E216A/E384R | 7.9fold increase in hydroxylase activity | Pseudonocardia autotrophica |
| 1.14.14.24 | V156S | 2- to 3fold increase in hydroxylase activity | Pseudonocardia autotrophica |
KM Value [mM]
| EC Number | KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|---|
| 1.14.14.24 | 0.0071 | - |
25-hydroxy-vitamin D3 | 30°C, pH not specified in the publication | Pseudonocardia autotrophica | |
| 1.14.14.24 | 0.0135 | - |
vitamin D3 | 30°C, pH not specified in the publication | Pseudonocardia autotrophica | |
| 1.14.14.24 | 0.032 | - |
vitamin D3 | mutant T70R/V156L/E216M/E384R, 30°C, pH not specified in the publication | Pseudonocardia autotrophica | |
Molecular Weight [Da]
| EC Number | Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
|---|---|---|---|---|
| 1.14.14.24 | 44368 | - |
x * 44368, calculated | Pseudonocardia autotrophica |
Organism
| EC Number | Organism | UniProt | Comment | Textmining |
|---|---|---|---|---|
| 1.14.14.24 | Pseudonocardia autotrophica | C4B644 | - |
- |
| 1.14.14.24 | Pseudonocardia autotrophica NBRC 12743 | C4B644 | - |
- |
Substrates and Products (Substrate)
| EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|---|
| 1.14.14.24 | vitamin D3 + O2 + NADPH + H+ | - |
Pseudonocardia autotrophica | 1alpha, 25-dihydroxy-vitamin D3 + NADP+ + H2O | major products are 25-hydroxy-vitamin D3 and 1alpha, 25-dihydroxy-vitamin D3, plus minor amounts of 26-hydroxy-vitamin D3 | ? | |
| 1.14.14.24 | vitamin D3 + O2 + NADPH + H+ | - |
Pseudonocardia autotrophica NBRC 12743 | 1alpha, 25-dihydroxy-vitamin D3 + NADP+ + H2O | major products are 25-hydroxy-vitamin D3 and 1alpha, 25-dihydroxy-vitamin D3, plus minor amounts of 26-hydroxy-vitamin D3 | ? | |
| 1.14.14.24 | vitamin D3 + O2 + NADPH + H+ | - |
Pseudonocardia autotrophica | 25-hydroxy-vitamin D3 + NADP+ + H2O | major products are 25-hydroxy-vitamin D3 and 1alpha, 25-dihydroxy-vitamin D3, plus minor amounts of 26-hydroxy-vitamin D3. Data indicate that Vdh first hydroxylates the C-25 position of vitamion D3 and subsequently hydroxylates the C1alpha position of 25-hydroxy-vitamin D3 | ? | |
| 1.14.14.24 | vitamin D3 + O2 + NADPH + H+ | - |
Pseudonocardia autotrophica NBRC 12743 | 25-hydroxy-vitamin D3 + NADP+ + H2O | major products are 25-hydroxy-vitamin D3 and 1alpha, 25-dihydroxy-vitamin D3, plus minor amounts of 26-hydroxy-vitamin D3. Data indicate that Vdh first hydroxylates the C-25 position of vitamion D3 and subsequently hydroxylates the C1alpha position of 25-hydroxy-vitamin D3 | ? | |
Subunits
| EC Number | Subunits | Comment | Organism |
|---|---|---|---|
| 1.14.14.24 | ? | x * 44368, calculated | Pseudonocardia autotrophica |
Synonyms
| EC Number | Synonyms | Comment | Organism |
|---|---|---|---|
| 1.14.14.24 | VDH | - |
Pseudonocardia autotrophica |
| 1.14.14.24 | vitamin D3 hydroxylase | - |
Pseudonocardia autotrophica |
Temperature Optimum [°C]
| EC Number | Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|---|
| 1.14.14.24 | 30 | - |
- |
Pseudonocardia autotrophica |
pH Optimum
| EC Number | pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|---|
| 1.14.14.24 | 7.5 | 8 | - |
Pseudonocardia autotrophica |
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