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
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1.13.11.3
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catecholate
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1,2-dioxygenase
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intradiol
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4-hydroxybenzoate
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3,4-dihydroxybenzoate
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beta-ketoadipate
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ring-cleavage
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gentisate
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beta-carboxy-cis,cis-muconate
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enol-lactone
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hydroxyquinol
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orville
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lipscomb
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fuscum
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cis-muconate
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4-methylcatechols
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3,5-di-tert-butylcatechol
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analysis
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environmental protection
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degradation
- 1.13.11.3
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catecholate
-
1,2-dioxygenase
-
intradiol
- 4-hydroxybenzoate
- 3,4-dihydroxybenzoate
-
beta-ketoadipate
-
ring-cleavage
- gentisate
- beta-carboxy-cis,cis-muconate
-
enol-lactone
- hydroxyquinol
-
orville
-
lipscomb
- fuscum
-
cis-muconate
- 4-methylcatechols
- 3,5-di-tert-butylcatechol
- analysis
- environmental protection
- degradation
Reaction
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
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Purification
Purification on EC 1.13.11.3 - protocatechuate 3,4-dioxygenase
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a highly active enzyme without nuclease contaminations is purified by Ni-NTA affinity chromatography and size exclusion chromatography for prolonged single-molecule fluorescence imaging
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ammonium sulfate, DEAE-Sepharose, octyl-Sepharose, Phenyl-sepharose, recombinant enzyme
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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
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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)
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native enzyme 296.8fold by ammonium sulfate fractionation, anion exchange and hydrophobic interaction chromatography
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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
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Q-Sepharose FF, ammonium sulfate, Sephacryl S-300HR, phenyl-superose HR5/5, Mono Q, type II protocatechuate 3,4-dioxygenase
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Q-Sepharose FF, ammonium sulfate, Sephacryl S-300HR, Phenyl-Superose, type I protocatechuate 3,4-dioxygenase
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Q-Sepharose FF, ammonium sulfate, Superdex 200, Fractogel, type II protocatechuate 3,4-dioxygenase
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