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Literature summary extracted from
- Quercetinase QueD of Streptomyces sp. FLA, a monocupin dioxygenase with a preference for nickel and cobalt (2008), Biochemistry, 47, 12185-12196.
Cloned(Commentary)
| EC Number | Cloned (Comment) | Organism |
|---|---|---|
| 1.13.11.24 | expression in Escherichia coli BL21(DE3) (pLysS, pET23a-queD) as a His6-tagged protein | Streptomyces sp. |
Protein Variants
| EC Number | Protein Variants | Comment | Organism |
|---|---|---|---|
| 1.13.11.24 | 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]
| EC Number | KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|---|
| 1.13.11.24 | 0.00096 | - |
quercetin | recombinant Co-QueD, 50 mM Tris-HCl buffer pH 8 | Streptomyces sp. |  |
| 1.13.11.24 | 0.00575 | - |
quercetin | recombinant Ni-QueD, 50 mM Tris-HCl buffer pH 8 | Streptomyces sp. | |
Metals/Ions
| EC Number | Metals/Ions | Comment | Organism | Structure |
|---|---|---|---|---|
| 1.13.11.24 | Co2+ | supplementing the cultures of strain FLA with CoCl2 results in 1.6fold higher quercetinase activity in crude extracts | Streptomyces sp. | |
| 1.13.11.24 | additional information | no increase in activity is observed when Mn2+, Fe2+, Cu2+, or Zn2+ is added to the culture medium | Streptomyces sp. | |
| 1.13.11.24 | Ni2+ | supplementing the cultures of strain FLA with NiCl2 results in 6.1fold higher quercetinase activity in crude extracts | Streptomyces sp. | |
Molecular Weight [Da]
| EC Number | Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
|---|---|---|---|---|
| 1.13.11.24 | 23040 | - |
calculated value of the hexahistidine-tagged QueD monomer | Streptomyces sp. |
| 1.13.11.24 | 23041 | - |
3 * 23041, gel permeation chromatography | Streptomyces sp. |
| 1.13.11.24 | 45000 | - |
SDS-PAGE | Streptomyces sp. |
| 1.13.11.24 | 65000 | - |
gel permeation chromatography, native Ni-QueD | Streptomyces sp. |
| 1.13.11.24 | 65100 | - |
gel permeation chromatography, native Co-QueD | Streptomyces sp. |
Natural Substrates/ Products (Substrates)
| EC Number | Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|---|
| 1.13.11.24 | quercetin + O2 | Streptomyces sp. | - |
2-protocatechoylphloroglucinolcarboxylate + CO | - |
? | |
| 1.13.11.24 | quercetin + O2 | Streptomyces sp. FLA / DSM 41951 | - |
2-protocatechoylphloroglucinolcarboxylate + CO | - |
? | |
Organism
| EC Number | Organism | UniProt | Comment | Textmining |
|---|---|---|---|---|
| 1.13.11.24 | Streptomyces sp. | - |
- |
- |
| 1.13.11.24 | Streptomyces sp. FLA / DSM 41951 | - |
- |
- |
Purification (Commentary)
| EC Number | Purification (Comment) | Organism |
|---|---|---|
| 1.13.11.24 | metal chelate affinity chromatography | Streptomyces sp. |
Specific Activity [micromol/min/mg]
| EC Number | Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
|---|---|---|---|---|
| 1.13.11.24 | 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. |
| 1.13.11.24 | 4 | - |
Fe-QueD | Streptomyces sp. |
| 1.13.11.24 | 28 | - |
Co-QueD | Streptomyces sp. |
| 1.13.11.24 | 144 | - |
Ni-QueD | Streptomyces sp. |
Substrates and Products (Substrate)
| EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|---|
| 1.13.11.24 | 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. | ? | - |
? | |
| 1.13.11.24 | 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 | ? | - |
? | |
| 1.13.11.24 | 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. | ? | - |
? | |
| 1.13.11.24 | 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 | ? | - |
? | |
| 1.13.11.24 | 3,5,7,4'-tetrahydroxyflavone + O2 | 43% of the activity with quercetin, Co-QueD, 29% of the activity with quercetin, Ni-QueD | Streptomyces sp. | ? | - |
? | |
| 1.13.11.24 | 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 | ? | - |
? | |
| 1.13.11.24 | 3,5,7-trihydroxyflavone + O2 | 13% of the activity with quercetin, Co-QueD, 16% of the activity with quercetin, Ni-QueD | Streptomyces sp. | ? | - |
? | |
| 1.13.11.24 | 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 | ? | - |
? | |
| 1.13.11.24 | 3,7,3',4'-tetrahydroxyflavone + O2 | 35% of the activity with quercetin, Co-QueD, 15% of the activity with quercetin, Ni-QueD | Streptomyces sp. | ? | - |
? | |
| 1.13.11.24 | quercetin + O2 | - |
Streptomyces sp. | 2-protocatechoylphloroglucinolcarboxylate + CO | - |
? | |
| 1.13.11.24 | quercetin + O2 | - |
Streptomyces sp. FLA / DSM 41951 | 2-protocatechoylphloroglucinolcarboxylate + CO | - |
? | |
Subunits
| EC Number | Subunits | Comment | Organism |
|---|---|---|---|
| 1.13.11.24 | dimer | SDS-PAGE | Streptomyces sp. |
| 1.13.11.24 | trimer | 3 * 23041, gel permeation chromatography | Streptomyces sp. |
Synonyms
| EC Number | Synonyms | Comment | Organism |
|---|---|---|---|
| 1.13.11.24 | QueD | - |
Streptomyces sp. |
| 1.13.11.24 | quercetinase | - |
Streptomyces sp. |
Temperature Optimum [°C]
| EC Number | Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|---|
| 1.13.11.24 | 40 | - |
- |
Streptomyces sp. |
Turnover Number [1/s]
| EC Number | Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|---|
| 1.13.11.24 | 7.6 | - |
quercetin | recombinant Co-QueD, 50 mM Tris-HCl buffer pH 8 | Streptomyces sp. | |
| 1.13.11.24 | 12.5 | - |
O2 | recombinant Co-QueD, 50 mM Tris-HCl buffer pH 8 | Streptomyces sp. | |
| 1.13.11.24 | 19 | - |
O2 | recombinant Ni-QueD, 50 mM Tris-HCl buffer pH 8 | Streptomyces sp. | |
| 1.13.11.24 | 40.1 | - |
quercetin | recombinant Ni-QueD, 50 mM Tris-HCl buffer pH 8 | Streptomyces sp. | |
pH Optimum
| EC Number | pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|---|
| 1.13.11.24 | 8 | - |
- |
Streptomyces sp. |
pH Range
| EC Number | pH Minimum | pH Maximum | Comment | Organism |
|---|---|---|---|---|
| 1.13.11.24 | 6 | 9 | 50 mM MES-Tris buffer | Streptomyces sp. |
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