Activating Compound | Comment | Organism | Structure |
---|---|---|---|
DTT | Co QueD: 110% activity remains after 15 min incubation with 1 mM reagent | Bacillus subtilis | |
EDTA | Co QueD: 110% activity remains after 15 min incubation with 1 mM reagent | Bacillus subtilis | |
NaN3 | Co QueD: 110% activity remains after 15 min incubation with 1 mM reagent, Mn QueD = 100% activity remains after 15 min incubation with 1 mM reagent | Bacillus subtilis | |
Sodium ascorbate | Co QueD: 110% activity remains after 15 min incubation with 1 mM reagent | Bacillus subtilis |
Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli BL21(DE3) | Bacillus subtilis |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
diethyldithiocarbamate | Co QueD: 90% activity remains after 15 min incubation with 1 mM reagent, Mn QueD: 84% activity remains after 15 min incubation with 1 mM reagent | Bacillus subtilis | |
DTT | Mn QueD: 96% activity remains after 15 min incubation with 1 mM reagent | Bacillus subtilis | |
EDTA | Mn QueD: 90% activity remains after 15 min incubation with 1 mM reagent | Bacillus subtilis | |
H2O2 | Co QueD: 93% activity remains after 15 min incubation with 1 mM reagent, Mn QueD: 82% activity remains after 15 min incubation with 1 mM reagent | Bacillus subtilis | |
kojic acid | Co QueD: 92% activity remains after 15 min incubation with 1 mM reagent, Mn QueD: 100% activity remains after 15 min incubation with 1 mM reagent | Bacillus subtilis | |
NaCN | Co QueD: 91% activity remains after 15 min incubation with 1 mM reagent, Mn QueD: 89% activity remains after 15 min incubation with 1 mM reagent | Bacillus subtilis | |
O-ethylxanthic acid | Co QueD: 69% activity remains after 15 min incubation with 1 mM reagent, Mn QueD: 100% activity remains after 15 min incubation with 1 mM reagent | Bacillus subtilis | |
Sodium ascorbate | Mn QueD: 80% activity remains after 15 min incubation with 1 mM reagent | Bacillus subtilis |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.004 | - |
quercetin | Mn quercetin 2,3-dioxygenase (1.8 atoms Mn/subunit) | Bacillus subtilis | |
0.0052 | - |
quercetin | Fe quercetin 2,3-dioxygenase | Bacillus subtilis | |
0.0075 | - |
quercetin | Co quercetin 2,3-dioxygenase (0.65 atoms Co/subunit) | Bacillus subtilis | |
0.079 | - |
oxygen | Co quercetin 2,3-dioxygenase (0.65 atoms Co/subunit) | Bacillus subtilis | |
0.09 | - |
oxygen | Mn quercetin 2,3-dioxygenase (1.8 atoms Mn/subunit) | Bacillus subtilis | |
0.15 | - |
oxygen | Fe quercetin 2,3-dioxygenase | Bacillus subtilis |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Co2+ | Co2+ salt addition increases the activity of quercetin 2,3-dioxygenase 24fold. The Escherichia coli cultures were grown at 37°C and 200 rpm for 6 h, induced with isopropyl beta-D-thiogalactopyanoside to a final concentraton of 50 mg/l in the presence of 10 microM CoCl2, and allow to grow additional 4 h at 25°C. The protein contains 0.65-0.8 atom of cobalt and 0.1 atom of iron per subunit. | Bacillus subtilis | |
Cu2+ | Cu2+ salt addition increases the activity of quercetin 2,3-dioxygenase 1.4fold. The Escherichia coli cultures were grown at 37°C and 200 rpm for 6 h, induced with isopropyl beta-D-thiogalactopyanoside to a final concentraton of 50 mg/l in the presence of 10 microM CuCl2, and allow to grow additional 4 h at 25°C. | Bacillus subtilis | |
Mn2+ | Mn2+ salt addition increases the activity of quercetin 2,3-dioxygenase 35fold. The Escherichia coli cultures were grown at 37°C and 200 rpm for 6 h, induced with isopropyl beta-D-thiogalactopyanoside to a final concentraton of 50 mg/l in the presence of 10 microM MnSO4, and allow to grow additional 4 h at 25°C. The protein containes 1.6-1.9 atoms of Mn/subunit. | Bacillus subtilis | |
additional information | Cd2+ does not increase the activity of quercetin 2,3-dioxygenase. The Escherichia coli cultures were grown at 37°C and 200 rpm for 6 h, induced with isopropyl beta-D-thiogalactopyanoside to a final concentraton of 50 mg/l in the presence of 10 microM CdCl2, and allow to grow additional 4 h at 25°C. | Bacillus subtilis | |
additional information | Fe2+ does not increase the activity of quercetin 2,3-dioxygenase. The Escherichia coli cultures were grown at 37°C and 200 rpm for 6 h, induced with isopropyl beta-D-thiogalactopyanoside to a final concentraton of 50 mg/l in the presence of 10 microM FeCl2, and allow to grow additional 4 h at 25°C. | Bacillus subtilis | |
additional information | Zn2+ does not increase the activity of quercetin 2,3-dioxygenase. The Escherichia coli cultures were grown at 37°C and 200 rpm for 6 h, induced with isopropyl beta-D-thiogalactopyanoside to a final concentraton of 50 mg/l in the presence of 10 microM ZnSO4, and allow to grow additional 4 h at 25°C. | Bacillus subtilis | |
Ni2+ | Ni2+ salt addition increases the activity of quercetin 2,3-dioxygenase 2.6fold. The Escherichia coli cultures were grown at 37°C and 200 rpm for 6 h, induced with isopropyl beta-D-thiogalactopyanoside to a final concentraton of 50 mg/l in the presence of 10 microM NiCl2, and allow to grow additional 4 h at 25°C. | Bacillus subtilis |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Bacillus subtilis | - |
- |
- |
Purification (Comment) | Organism |
---|---|
DEAE-Sephacel column equilibrated with 50 mM Tris.HCl, pH 7.5, and eluted with a NaCl gradient (0-600 mM). Ultrogel ACA 34 column, eluted with 50 mM Tris.HCl, pH 7.5, and 100 mM Nacl. DEAE-Sepharose column equilibrated with 50 mM Tris.HCl, pH 7,5, eluted with a gradient of NaCl (100-500 mM). | Bacillus subtilis |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
quercetin + O2 | - |
Bacillus subtilis | 2-(3,4-dihydroxybenzoyloxy)-4,6-dihydroxybenzoate + CO + H+ | - |
? |
Synonyms | Comment | Organism |
---|---|---|
QueD | - |
Bacillus subtilis |
quercetinase | - |
Bacillus subtilis |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.65 | - |
quercetin | Fe quercetin 2,3-dioxygenase | Bacillus subtilis | |
6.7 | - |
quercetin | Co quercetin 2,3-dioxygenase (0.65 atoms Co/subunit) | Bacillus subtilis | |
25 | - |
quercetin | Mn quercetin 2,3-dioxygenase (1.8 atoms Mn/subunit) | Bacillus subtilis |
pH Minimum | pH Maximum | Comment | Organism |
---|---|---|---|
additional information | - |
pH profiles are determined spectrophotometrically on a Hp 8453 photodiode array spectrophotometer | Bacillus subtilis |
Organism | Comment | pI Value Maximum | pI Value |
---|---|---|---|
Bacillus subtilis | pKa determined from the Vmax profile is 5.6. The pKa determined from the Vmax/KM pH profile is 5.8. | - |
additional information |