Cloned (Comment) | Organism |
---|---|
expression in Escherichia coli BL21(DE3) (pLysS, pET23a-queD) as a His6-tagged protein | Streptomyces sp. |
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 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 | 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 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 | 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 | UniProt | Comment | Textmining |
---|---|---|---|
Streptomyces sp. | - |
- |
- |
Streptomyces sp. FLA / DSM 41951 | - |
- |
- |
Purification (Comment) | Organism |
---|---|
metal chelate affinity chromatography | Streptomyces sp. |
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 | 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 | Comment | Organism |
---|---|---|
dimer | SDS-PAGE | Streptomyces sp. |
trimer | 3 * 23041, gel permeation chromatography | Streptomyces sp. |
Synonyms | Comment | Organism |
---|---|---|
QueD | - |
Streptomyces sp. |
quercetinase | - |
Streptomyces sp. |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
40 | - |
- |
Streptomyces sp. |
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 Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
8 | - |
- |
Streptomyces sp. |
pH Minimum | pH Maximum | Comment | Organism |
---|---|---|---|
6 | 9 | 50 mM MES-Tris buffer | Streptomyces sp. |