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Literature summary for 1.13.11.24 extracted from

  • Merkens, H.; Kappl, R.; Jakob, R.P.; Schmid, F.X.; Fetzner, S.
    Quercetinase QueD of Streptomyces sp. FLA, a monocupin dioxygenase with a preference for nickel and cobalt (2008), Biochemistry, 47, 12185-12196.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
expression in Escherichia coli BL21(DE3) (pLysS, pET23a-queD) as a His6-tagged protein Streptomyces sp.

Protein Variants

Protein Variants Comment Organism
E76A site-directed mutagenesis of Ni-QueD, exhibits marginal quercetinase activity of 0.014 unit/mg, corresponding to a more than 10000fold decrease in specific activity, Glu76 of QueD is part of the strictly conserved cupin motif and thus is assumed to be a ligand to the metal center Streptomyces sp.

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.00096
-
quercetin recombinant Co-QueD, 50 mM Tris-HCl buffer pH 8 Streptomyces sp.
0.00575
-
quercetin recombinant Ni-QueD, 50 mM Tris-HCl buffer pH 8 Streptomyces sp.

Metals/Ions

Metals/Ions Comment Organism Structure
Co2+ supplementing the cultures of strain FLA with CoCl2 results in 1.6fold higher quercetinase activity in crude extracts Streptomyces sp.
additional information no increase in activity is observed when Mn2+, Fe2+, Cu2+, or Zn2+ is added to the culture medium Streptomyces sp.
Ni2+ supplementing the cultures of strain FLA with NiCl2 results in 6.1fold higher quercetinase activity in crude extracts Streptomyces sp.

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
23040
-
calculated value of the hexahistidine-tagged QueD monomer Streptomyces sp.
23041
-
3 * 23041, gel permeation chromatography Streptomyces sp.
45000
-
SDS-PAGE Streptomyces sp.
65000
-
gel permeation chromatography, native Ni-QueD Streptomyces sp.
65100
-
gel permeation chromatography, native Co-QueD Streptomyces sp.

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
quercetin + O2 Streptomyces sp.
-
2-protocatechoylphloroglucinolcarboxylate + CO
-
?
quercetin + O2 Streptomyces sp. FLA / DSM 41951
-
2-protocatechoylphloroglucinolcarboxylate + CO
-
?

Organism

Organism UniProt Comment Textmining
Streptomyces sp.
-
-
-
Streptomyces sp. FLA / DSM 41951
-
-
-

Purification (Commentary)

Purification (Comment) Organism
metal chelate affinity chromatography Streptomyces sp.

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
additional information
-
high specific QueD activity is found in crude cell extracts when the growth medium is supplemented with NiCl2 or CoCl2, but not when Mn2+, Fe2+, Cu2+, or Zn2+ is added, the activity of the purified QueD proteins depends strongly on the nature of the metal ion cofactor Streptomyces sp.
4
-
Fe-QueD Streptomyces sp.
28
-
Co-QueD Streptomyces sp.
144
-
Ni-QueD Streptomyces sp.

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3,5,7,2',4'-pentahydroxyflavone + O2 5.5% of the activity with quercetin, Co-QueD, 0.9% of the activity with quercetin, Ni-QueD Streptomyces sp. ?
-
?
3,5,7,2',4'-pentahydroxyflavone + O2 5.5% of the activity with quercetin, Co-QueD, 0.9% of the activity with quercetin, Ni-QueD Streptomyces sp. FLA / DSM 41951 ?
-
?
3,5,7,3',4',5'-hexahydroxyflavone + O2 77% of the activity with quercetin, Co-QueD, 46% of the activity with quercetin, Ni-QueD Streptomyces sp. ?
-
?
3,5,7,3',4',5'-hexahydroxyflavone + O2 77% of the activity with quercetin, Co-QueD, 46% of the activity with quercetin, Ni-QueD Streptomyces sp. FLA / DSM 41951 ?
-
?
3,5,7,4'-tetrahydroxyflavone + O2 43% of the activity with quercetin, Co-QueD, 29% of the activity with quercetin, Ni-QueD Streptomyces sp. ?
-
?
3,5,7,4'-tetrahydroxyflavone + O2 43% of the activity with quercetin, Co-QueD, 29% of the activity with quercetin, Ni-QueD Streptomyces sp. FLA / DSM 41951 ?
-
?
3,5,7-trihydroxyflavone + O2 13% of the activity with quercetin, Co-QueD, 16% of the activity with quercetin, Ni-QueD Streptomyces sp. ?
-
?
3,5,7-trihydroxyflavone + O2 13% of the activity with quercetin, Co-QueD, 16% of the activity with quercetin, Ni-QueD Streptomyces sp. FLA / DSM 41951 ?
-
?
3,7,3',4'-tetrahydroxyflavone + O2 35% of the activity with quercetin, Co-QueD, 15% of the activity with quercetin, Ni-QueD Streptomyces sp. ?
-
?
quercetin + O2
-
Streptomyces sp. 2-protocatechoylphloroglucinolcarboxylate + CO
-
?
quercetin + O2
-
Streptomyces sp. FLA / DSM 41951 2-protocatechoylphloroglucinolcarboxylate + CO
-
?

Subunits

Subunits Comment Organism
dimer SDS-PAGE Streptomyces sp.
trimer 3 * 23041, gel permeation chromatography Streptomyces sp.

Synonyms

Synonyms Comment Organism
QueD
-
Streptomyces sp.
quercetinase
-
Streptomyces sp.

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
40
-
-
Streptomyces sp.

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
7.6
-
quercetin recombinant Co-QueD, 50 mM Tris-HCl buffer pH 8 Streptomyces sp.
12.5
-
O2 recombinant Co-QueD, 50 mM Tris-HCl buffer pH 8 Streptomyces sp.
19
-
O2 recombinant Ni-QueD, 50 mM Tris-HCl buffer pH 8 Streptomyces sp.
40.1
-
quercetin recombinant Ni-QueD, 50 mM Tris-HCl buffer pH 8 Streptomyces sp.

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
8
-
-
Streptomyces sp.

pH Range

pH Minimum pH Maximum Comment Organism
6 9 50 mM MES-Tris buffer Streptomyces sp.