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Literature summary for 1.14.14.24 extracted from

  • Fujii, Y.; Kabumoto, H.; Nishimura, K.; Fujii, T.; Yanai, S.; Takeda, K.; Tamura, N.; Arisawa, A.; Tamura, T.
    Purification, characterization, and directed evolution study of a vitamin D3 hydroxylase from Pseudonocardia autotrophica (2009), Biochem. Biophys. Res. Commun., 385, 170-175.
    View publication on PubMed

Activating Compound

Activating Compound Comment Organism Structure
KCl or NaCl, required. Optimum concentration 100 mM Pseudonocardia autotrophica
NaCl or KCl, required. Optimum concentration 100 mM Pseudonocardia autotrophica
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)

Cloned (Comment) Organism
expression in Escherichia coli Pseudonocardia autotrophica

Protein Variants

Protein Variants Comment Organism
E216A 2- to 3fold increase in hydroxylase activity Pseudonocardia autotrophica
E384R 2- to 3fold increase in hydroxylase activity Pseudonocardia autotrophica
T70R 2- to 3fold increase in hydroxylase activity Pseudonocardia autotrophica
T70R/V156L/E216M/E384R 21fold increase in hydroxylase activity Pseudonocardia autotrophica
T70R/V156S/E216A/E384R 7.9fold increase in hydroxylase activity Pseudonocardia autotrophica
V156S 2- to 3fold increase in hydroxylase activity Pseudonocardia autotrophica

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.0071
-
25-hydroxy-vitamin D3 30°C, pH not specified in the publication Pseudonocardia autotrophica
0.0135
-
vitamin D3 30°C, pH not specified in the publication Pseudonocardia autotrophica
0.032
-
vitamin D3 mutant T70R/V156L/E216M/E384R, 30°C, pH not specified in the publication Pseudonocardia autotrophica

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
44368
-
x * 44368, calculated Pseudonocardia autotrophica

Organism

Organism UniProt Comment Textmining
Pseudonocardia autotrophica C4B644
-
-
Pseudonocardia autotrophica NBRC 12743 C4B644
-
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
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 ?
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 ?
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 ?
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

Subunits Comment Organism
? x * 44368, calculated Pseudonocardia autotrophica

Synonyms

Synonyms Comment Organism
VDH
-
Pseudonocardia autotrophica
vitamin D3 hydroxylase
-
Pseudonocardia autotrophica

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
30
-
-
Pseudonocardia autotrophica

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.5 8
-
Pseudonocardia autotrophica