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Literature summary extracted from

  • Almabruk, K.H.; Asamizu, S.; Chang, A.; Varghese, S.G.; Mahmud, T.
    The alpha-ketoglutarate/Fe(II)-dependent dioxygenase VldW is responsible for the formation of validamycin B (2012), Chembiochem, 13, 2209-2211.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.14.11.52 inserted into an expression vector pRSET B to give pTMS005. The plasmid is then used to transform Escherichia coli BL21(DE3)pLysS Streptomyces hygroscopicus subsp. limoneus

Inhibitors

EC Number Inhibitors Comment Organism Structure
1.14.11.52 Ni2+ atalytic activity is affected by Ni2+. A significant reduction of activity is observed when the protein is purified using Ni-NTA column. The activity can be restored by dialysis of the protein in a buffer solution containing 0.1 mM EDTA, followed by the addition of 0.2 mM of Fe2+ to the protein solution Streptomyces hygroscopicus subsp. limoneus

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.14.11.52 0.019
-
2-oxoglutarate pH 7.2, 30°C, obtained from Hanes-Woolf plot Streptomyces hygroscopicus subsp. limoneus
1.14.11.52 0.303
-
validamycin A pH 7.2, 30°C, obtained from Hanes-Woolf plot Streptomyces hygroscopicus subsp. limoneus

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
1.14.11.52 Fe2+ required Streptomyces hygroscopicus subsp. limoneus

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
1.14.11.52 43400
-
x * 43400, recombinantly His6-tagged protein, SDS-PAGE Streptomyces hygroscopicus subsp. limoneus

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
1.14.11.52 validamycin A + 2-oxoglutarate + O2 Streptomyces hygroscopicus subsp. limoneus
-
validamycin B + succinate + CO2
-
?

Organism

EC Number Organism UniProt Comment Textmining
1.14.11.52 Streptomyces hygroscopicus subsp. limoneus Q15JG7
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.14.11.52
-
Streptomyces hygroscopicus subsp. limoneus

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.14.11.52 additional information no products are observed when validamine, validamine 7-phosphate, or validoxylamine A 7'-phosphate are used as substrate Streptomyces hygroscopicus subsp. limoneus ?
-
?
1.14.11.52 validamycin A + 2-oxoglutarate + O2
-
Streptomyces hygroscopicus subsp. limoneus validamycin B + succinate + CO2
-
?
1.14.11.52 validamycin A + 2-oxoglutarate + O2 validamycin A i.e. (1R,2R,3S,4S,6R)-2,3-dihydroxy-6-(hydroxymethyl)-4-[[(1S,4R,5S,6S)-4,5,6-trihydroxy-3-(hydroxymethyl)cyclohex-2-en-1-yl]amino]cyclohexyl beta-D-glucopyranoside Streptomyces hygroscopicus subsp. limoneus validamycin B + succinate + CO2 validamycin B i.e. (1R,2R,3S,4S,5R,6S)-2,3,5-trihydroxy-6-(hydroxymethyl)-4-8[[(1S,4R,5S,6S)-4,5,6-trihydroxy-3-(hydroxymethyl)cyclohex-2-en-1-yl]amino]cyclohexyl beta-D-glucopyranoside ?
1.14.11.52 validoxylamine A + 2-oxoglutarate + O2 the enzyme is also able to convert validoxylamine A to its hydroxylated derivative, albeit much less efficiently than the conversion of validamycin A to validamycin B Streptomyces hygroscopicus subsp. limoneus ?
-
?

Subunits

EC Number Subunits Comment Organism
1.14.11.52 ? x * 43400, recombinantly His6-tagged protein, SDS-PAGE Streptomyces hygroscopicus subsp. limoneus

Synonyms

EC Number Synonyms Comment Organism
1.14.11.52 VldW
-
Streptomyces hygroscopicus subsp. limoneus

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.14.11.52 0.97
-
2-oxoglutarate pH 7.2, 30°C, obtained from Hanes-Woolf plot Streptomyces hygroscopicus subsp. limoneus
1.14.11.52 0.97
-
validamycin A pH 7.2, 30°C, obtained from Hanes-Woolf plot Streptomyces hygroscopicus subsp. limoneus

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.14.11.52 7.2
-
the enzyme is most active in potassium phosphate buffer Streptomyces hygroscopicus subsp. limoneus

pH Range

EC Number pH Minimum pH Maximum Comment Organism
1.14.11.52 6.5 8 pH 6.5: 65% of maximal activity, pH 8.0: about 70% of maximal activity Streptomyces hygroscopicus subsp. limoneus