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Literature summary extracted from

  • Schaab, M.R.; Barney, B.M.; Francisco, W.A.
    Kinetic and spectroscopic studies on the quercetin 2,3-dioxygenase from Bacillus subtilis (2006), Biochemistry, 45, 1009-1016.
    View publication on PubMed

Activating Compound

EC Number Activating Compound Comment Organism Structure
1.13.11.24 DTT Co QueD: 110% activity remains after 15 min incubation with 1 mM reagent Bacillus subtilis
1.13.11.24 EDTA Co QueD: 110% activity remains after 15 min incubation with 1 mM reagent Bacillus subtilis
1.13.11.24 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
1.13.11.24 Sodium ascorbate Co QueD: 110% activity remains after 15 min incubation with 1 mM reagent Bacillus subtilis

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.13.11.24 expressed in Escherichia coli BL21(DE3) Bacillus subtilis

Inhibitors

EC Number Inhibitors Comment Organism Structure
1.13.11.24 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
1.13.11.24 DTT Mn QueD: 96% activity remains after 15 min incubation with 1 mM reagent Bacillus subtilis
1.13.11.24 EDTA Mn QueD: 90% activity remains after 15 min incubation with 1 mM reagent Bacillus subtilis
1.13.11.24 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
1.13.11.24 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
1.13.11.24 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
1.13.11.24 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
1.13.11.24 Sodium ascorbate Mn QueD: 80% activity remains after 15 min incubation with 1 mM reagent Bacillus subtilis

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.13.11.24 0.004
-
 quercetin Mn quercetin 2,3-dioxygenase (1.8 atoms Mn/subunit) Bacillus subtilis
1.13.11.24 0.0052
-
 quercetin Fe quercetin 2,3-dioxygenase Bacillus subtilis
1.13.11.24 0.0075
-
 quercetin Co quercetin 2,3-dioxygenase (0.65 atoms Co/subunit) Bacillus subtilis
1.13.11.24 0.079
-
 oxygen Co quercetin 2,3-dioxygenase (0.65 atoms Co/subunit) Bacillus subtilis
1.13.11.24 0.09
-
 oxygen Mn quercetin 2,3-dioxygenase (1.8 atoms Mn/subunit) Bacillus subtilis
1.13.11.24 0.15
-
 oxygen Fe quercetin 2,3-dioxygenase Bacillus subtilis

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
1.13.11.24 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
1.13.11.24 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
1.13.11.24 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
1.13.11.24 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
1.13.11.24 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
1.13.11.24 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
1.13.11.24 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

EC Number Organism UniProt Comment Textmining
1.13.11.24 Bacillus subtilis
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.13.11.24 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)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.13.11.24 quercetin + O2
-
 Bacillus subtilis 2-(3,4-dihydroxybenzoyloxy)-4,6-dihydroxybenzoate + CO + H+
-
 ?

Synonyms

EC Number Synonyms Comment Organism
1.13.11.24 QueD
-
 Bacillus subtilis
1.13.11.24 quercetinase
-
 Bacillus subtilis

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.13.11.24 0.65
-
 quercetin Fe quercetin 2,3-dioxygenase Bacillus subtilis
1.13.11.24 6.7
-
 quercetin Co quercetin 2,3-dioxygenase (0.65 atoms Co/subunit) Bacillus subtilis
1.13.11.24 25
-
 quercetin Mn quercetin 2,3-dioxygenase (1.8 atoms Mn/subunit) Bacillus subtilis

pH Range

EC Number pH Minimum pH Maximum Comment Organism
1.13.11.24 additional information
-
 pH profiles are determined spectrophotometrically on a Hp 8453 photodiode array spectrophotometer Bacillus subtilis

pI Value

EC Number Organism Comment pI Value Maximum pI Value
1.13.11.24 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