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Literature summary for 1.14.11.52 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
Search for more literature for 1.14.11.52

Cloned(Commentary)

Cloned (Comment) Organism
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

Inhibitors Comment Organism Structure
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]

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

Metals/Ions

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

Molecular Weight [Da]

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

Natural Substrates/ Products (Substrates)

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

Organism

Organism UniProt Comment Textmining
Streptomyces hygroscopicus subsp. limoneus Q15JG7
-
-

Purification (Commentary)

Purification (Comment) Organism
-
 Streptomyces hygroscopicus subsp. limoneus

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
additional information no products are observed when validamine, validamine 7-phosphate, or validoxylamine A 7'-phosphate are used as substrate Streptomyces hygroscopicus subsp. limoneus ?
-
 ?
validamycin A + 2-oxoglutarate + O2
-
 Streptomyces hygroscopicus subsp. limoneus validamycin B + succinate + CO2
-
 ?
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 ?
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

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

Synonyms

Synonyms Comment Organism
VldW
-
 Streptomyces hygroscopicus subsp. limoneus

Turnover Number [1/s]

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

pH Optimum

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

pH Range

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