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Literature summary for 1.13.11.24 extracted from
- Kinetic and spectroscopic studies on the quercetin 2,3-dioxygenase from Bacillus subtilis (2006), Biochemistry, 45, 1009-1016.
Activating Compound
| 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(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| expressed in Escherichia coli BL21(DE3) | Bacillus subtilis |
Inhibitors
| 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 [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
| 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
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Bacillus subtilis | - |
- |
- |
Purification (Commentary)
| 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 and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| quercetin + O2 | - |
Bacillus subtilis | 2-(3,4-dihydroxybenzoyloxy)-4,6-dihydroxybenzoate + CO + H+ | - |
? | |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| QueD | - |
Bacillus subtilis |
| quercetinase | - |
Bacillus subtilis |
Turnover Number [1/s]
| 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 Range
| pH Minimum | pH Maximum | Comment | Organism |
|---|---|---|---|
| additional information | - |
pH profiles are determined spectrophotometrically on a Hp 8453 photodiode array spectrophotometer | Bacillus subtilis |
pI Value
| 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 |
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