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1.13.11.3: protocatechuate 3,4-dioxygenase

This is an abbreviated version!
For detailed information about protocatechuate 3,4-dioxygenase, go to the full flat file.

Word Map on EC 1.13.11.3

Reaction

3,4-dihydroxybenzoate
+
O2
=
3-carboxy-cis,cis-muconate

Synonyms

3,4-PCase, 3,4-PCD, 3,4-PCDase, 3,4-POD, EC 1.13.1.3, EC 1.99.2.3, More, oxygenase, protocatechuate 3,4-di-, P3,4DO, P3,4O enzyme, P34O, PCA 3,4-dioxygenase, PcaG, PcaH, PcaHG, PCD, protocatchetuate 3,4-dioxygenase, protocatechuate 3,4-dioxygenase, protocatechuate oxygenase, protocatechuic 3,4-dioxygenase, protocatechuic 3,4-oxygenase, protocatechuic acid oxidase

ECTree

     1 Oxidoreductases
         1.13 Acting on single donors with incorporation of molecular oxygen (oxygenases)
             1.13.11 With incorporation of two atoms of oxygen
                1.13.11.3 protocatechuate 3,4-dioxygenase

Purification

Purification on EC 1.13.11.3 - protocatechuate 3,4-dioxygenase

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PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
a highly active enzyme without nuclease contaminations is purified by Ni-NTA affinity chromatography and size exclusion chromatography for prolonged single-molecule fluorescence imaging
-
ammonium sulfate, DEAE-cellulose
-
ammonium sulfate, DEAE-cellulose, Bio-gel agarose, DEAE-cellulose
-
ammonium sulfate, DEAE-cellulose, Sephadex G-200
-
ammonium sulfate, DEAE-Sepharose, octyl-Sepharose, Phenyl-sepharose, recombinant enzyme
-
ammonium sulfate, DEAE-Sepharose, Phenyl-Sepharose
-
ammonium sulfate, Q-Sepharose, partial purification
-
ammonium sulfate, Sephacryl S 300, DEAE-Sephadex A50
-
cells harvested by centrifugation, washed twice with 50 mM sodium phosphate buffer, pH 7.0 containing 10% glycerol, disruption with solicitor, centrifuged, supernatant stored at -20°C
-
enzymes from Escherichia coli: DEAE-Sepharose Fast Flow column (5.5 x 19 cm), Phenyl-Sepharose CL-4B column (4.5 x 22.5 cm), Sephacryl S-300 column (3 x 97.5 cm)
-
enzymes from Pseudomonas florescens
-
native enzyme 296.8fold by ammonium sulfate fractionation, anion exchange and hydrophobic interaction chromatography
-
Ni-affinity column chromatography
protocol for the expression and purification of highly active Pseudomonas putida protocatechuate-3,4-dioxygenase with no detectable nuclease contamination. Purification by nickel affinity chromatography, elution in 2 steps of imidazole concentrations, chromatography fractions are analyzed by SDS-PAGE, fractions of nearly pure enzyme are concentrated and further purified by size exclusion chromatography
-
Q-Sepharose FF, ammonium sulfate, Sephacryl S-300HR, phenyl-superose HR5/5, Mono Q, type II protocatechuate 3,4-dioxygenase
-
Q-Sepharose FF, ammonium sulfate, Sephacryl S-300HR, Phenyl-Superose, type I protocatechuate 3,4-dioxygenase
-
Q-Sepharose FF, ammonium sulfate, Superdex 200, Fractogel, type II protocatechuate 3,4-dioxygenase
-
recombinant enzyme
-