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Information on EC 1.13.11.3 - protocatechuate 3,4-dioxygenase
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1.13.11.3 protocatechuate 3,4-dioxygenaseIUBMB Comments
Requires Fe3+. The enzyme, which participates in the degradation of aromatic compounds, catalyses the intradiol addition of both oxygen atoms from molecular oxygen, resulting in ortho-cleavage of the aromatic ring. The type of cleavage leads to mineralization via the intermediate 3-oxoadipate.
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Word Map
- 1.13.11.3
-
catecholate
-
1,2-dioxygenase
-
intradiol
- p-hydroxybenzoate
- 3,4-dihydroxybenzoate
-
beta-ketoadipate
-
vanillic
-
ring-cleavage
- gentisate
- beta-carboxy-cis,cis-muconate
-
lipscomb
-
fuscum
-
enol-lactone
-
orville
- 3,5-di-tert-butylcatechol
- 4-methylcatechols
- hydroxyquinol
- environmental protection
- analysis
- degradation
The enzyme appears in viruses and cellular organisms
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, 
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SYNONYM 
ORGANISM
UNIPROT
COMMENTARY 
LITERATURE
3,4-PCD
EC 1.13.1.3
-
-
formerly
-
EC 1.99.2.3
-
-
formerly
-
oxygenase, protocatechuate 3,4-di-
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-
-
-
P3,4DO
P34O
PcaG
PcaH
PcaHG
protocatechuate 3,4-dioxygenase
protocatechuate oxygenase
-
-
-
-
protocatechuic 3,4-dioxygenase
-
-
-
-
protocatechuic 3,4-oxygenase
-
-
-
-
protocatechuic acid oxidase
-
-
-
-
additional information
additional information
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Experiments with mutants where the pcaG genes contain variations in repeat sequences capable of producing a selectable phenotype following a specific deletion give following results: As a lower limit a sequence repition of 8 bp or greater in length is required for deletion frequencies to rise significantly above 1.5 x 10-8. Deletions occurr with a frequency of 7.2 x 10-8 in strains containing repetitions of 12 bp in length separated by 30 bp
additional information
-
the genetic locus ncg12314-ncg12315 encodes a putative protocatechuate 3,4-dioxygenase
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REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT
LITERATURE
3,4-dihydroxybenzoate + O2 = 3-carboxy-cis,cis-muconate
-
-
-
-
3,4-dihydroxybenzoate + O2 = 3-carboxy-cis,cis-muconate
proposed mechanism based on Mössbauer, EPR and inhibition kinetic data
-
3,4-dihydroxybenzoate + O2 = 3-carboxy-cis,cis-muconate
proposed reaction mechanism
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3,4-dihydroxybenzoate + O2 = 3-carboxy-cis,cis-muconate
examining a distorted trigonal-bipyramidal geometry observed for the non-heme iron center in protocatechuate 3,4-dioxygenase by utilizing a sterically hindered iron salen complex gives following results: the extent of a structural change of the iron center from a preferred square-pyramidal to a distorted trigonal-bipyramidal geometry varies with the external ligand that is bound in the order Cl <
-
3,4-dihydroxybenzoate + O2 = 3-carboxy-cis,cis-muconate
the computational (hybrid density functional method B3LYP) results of the catalytic cycle of the intradiol dioxygenases suggest: (1)binging of the substrate as a dianion (2) binding of dioxygen to the metal aided by an electron transfer from the substrate to O2 (3) formation of a bridging peroxo intermediate and its conformational change, which opens the coordination site trans to His462, (4) binding of a neutral XOH ligand (H2O or Tyr447) at the open site, (5) proton transfer from XOH to the neighboring peroxo ligand yielding the hydroperoxo intermediate, (6) a Criegee rearrangment leading to the anhydride intermediate (the criegee mechanism requires an in-plane oriantation of the involved two oxygen and two carbon atoms. Under some conditions homolytic O-O bond cleavage might compete with the Criegee rearangment.) and (7) hydrolysis of the anhydride to the final acyclic product.
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REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
oxidation
-
-
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redox reaction
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-
-
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reduction
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-
-
-
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PATHWAY SOURCE
PATHWAYS
KEGG
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SYSTEMATIC NAME
IUBMB Comments
protocatechuate:oxygen 3,4-oxidoreductase (decyclizing)
Requires Fe3+. The enzyme, which participates in the degradation of aromatic compounds, catalyses the intradiol addition of both oxygen atoms from molecular oxygen, resulting in ortho-cleavage of the aromatic ring. The type of cleavage leads to mineralization via the intermediate 3-oxoadipate.
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CAS REGISTRY NUMBER
COMMENTARY
9029-47-4
-
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SUBSTRATE
PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
3,4-dihydroxyphenylalanine + O2
?
Nocardia erythropolis
-
at 10.7% the rate of protocatechuic acid oxidation
-
-
?
3-(3,4-dihydroxyphenyl)propionate + O2
?
-
protocatechuate 3,4-dioxygenase II
-
-
?
5-fluoro-protocatechuic acid + O2
5-fluoro-3-carboxy-cis,cis-muconate
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at 2.1% the rate of protocatechuic acid oxidation
-
?
6-chloro-protocatechuate + O2
6-chloro-3-carboxy-cis,cis-muconate
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at 4.3% the rate of protocatechuic acid oxidation
-
?
2,3-dihydroxybenzoate + O2
?
23.6% activity compared to 3,4-dihydroxybenzoate
-
-
?
2,3-dihydroxybenzoate + O2
?
23.6% of the activity with protocatechuate for the free enzyme, 149.3% for the calcium alginate-immobilized enzyme, and 119.8% for the glyoxyl agarose-immobilized enzyme
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-
?
2,4-dihydroxybenzoate + O2
?
43.5% activity compared to 3,4-dihydroxybenzoate
-
-
?
2,4-dihydroxybenzoate + O2
?
43.5% of the activity with protocatechuate for the free enzyme, 96.5% for the calcium alginate-immobilized enzyme, and 65.6% for the glyoxyl agarose-immobilized enzyme
-
-
?
2,5-dihydroxybenzoate + O2
?
33.46% activity compared to 3,4-dihydroxybenzoate
-
-
?
2,5-dihydroxybenzoate + O2
?
33.5% of the activity with protocatechuate for the free enzyme, 158.6% for the calcium alginate-immobilized enzyme, and 72.1% for the glyoxyl agarose-immobilized enzyme
-
-
?
2,6-dihydroxybenzoate + O2
?
30.5% of the activity with protocatechuate for the free enzyme, 90.4% for the calcium alginate-immobilized enzyme, and 36.2% for the glyoxyl agarose-immobilized enzyme
-
-
?
2,6-dihydroxybenzoate + O2
?
30.51% activity compared to 3,4-dihydroxybenzoate
-
-
?
3,4-dihydroxybenzoate + O2
3-carboxy-cis,cis-muconate
-
-
-
?
3,4-dihydroxybenzoate + O2
3-carboxy-cis,cis-muconate
-
-
-
?
3,4-dihydroxybenzoate + O2
3-carboxy-cis,cis-muconate
-
-
-
?
3,4-dihydroxybenzoate + O2
3-carboxy-cis,cis-muconate
-
-
-
-
3,4-dihydroxybenzoate + O2
3-carboxy-cis,cis-muconate
-
-
-
?
3,4-dihydroxybenzoate + O2
3-carboxy-cis,cis-muconate
-
-
-
?
3,4-dihydroxybenzoate + O2
3-carboxy-cis,cis-muconate
-
-
-
-
?
3,4-dihydroxybenzoate + O2
3-carboxy-cis,cis-muconate
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absolute requirement for vicinal hydroxyl groups in the 3- and 4-position
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ir
3,4-dihydroxybenzoate + O2
3-carboxy-cis,cis-muconate
-
very low activity with 2-chloro-protocatechuate
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?
3,4-dihydroxybenzoate + O2
3-carboxy-cis,cis-muconate
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very low activity with 5-bromo-protocatechuate
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?
3,4-dihydroxybenzoate + O2
3-carboxy-cis,cis-muconate
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very low activity with 5-chloro-protocatechuate
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?
3,4-dihydroxybenzoate + O2
3-carboxy-cis,cis-muconate
-
-
-
-
3,4-dihydroxybenzoate + O2
3-carboxy-cis,cis-muconate
-
-
-
?
3,4-dihydroxybenzoate + O2
3-carboxy-cis,cis-muconate
-
-
-
?
3,4-dihydroxybenzoate + O2
3-carboxy-cis,cis-muconate
-
methylene blue cannot replace O2 as electron acceptor
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ir
3,4-dihydroxybenzoate + O2
3-carboxy-cis,cis-muconate
Nocardia erythropolis
-
enzyme is active on a wide range of o-dihydroxyphenyl compounds
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?
3,4-dihydroxybenzoate + O2
3-carboxy-cis,cis-muconate
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very low activity with: 3,4-dihydroxyphenylacetic acid
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?
3,4-dihydroxybenzoate + O2
3-carboxy-cis,cis-muconate
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very low activity with 3,4-dihydroxymandelic acid
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?
3,4-dihydroxybenzoate + O2
3-carboxy-cis,cis-muconate
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very low activity with 4'-methylcatechol
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?
3,4-dihydroxybenzoate + O2
3-carboxy-cis,cis-muconate
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very low activity with: 3,4-dihydroxyphenylacetic acid
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?
3,4-dihydroxybenzoate + O2
3-carboxy-cis,cis-muconate
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very low activity with 3,4-dihydroxymandelic acid
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?
3,4-dihydroxybenzoate + O2
3-carboxy-cis,cis-muconate
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very low activity with 4'-methylcatechol
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?
3,4-dihydroxybenzoate + O2
3-carboxy-cis,cis-muconate
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no other substrate found
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ir
3,4-dihydroxybenzoate + O2
3-carboxy-cis,cis-muconate
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no other substrate found
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ir
3,4-dihydroxybenzoate + O2
3-carboxy-cis,cis-muconate
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-
-
-
?
3,4-dihydroxybenzoate + O2
3-carboxy-cis,cis-muconate
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i.e. protocatechuate
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?
3,4-dihydroxybenzoate + O2
3-carboxy-cis,cis-muconate
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?
3,4-dihydroxybenzoate + O2
3-carboxy-cis,cis-muconate
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specific for protocatechuate
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?
3,4-dihydroxybenzoate + O2
3-carboxy-cis,cis-muconate
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?
3,4-dihydroxybenzoate + O2
3-carboxy-cis,cis-muconate
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-
-
?
3,4-dihydroxybenzoate + O2
3-carboxy-cis,cis-muconate
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-
-
?
3,4-dihydroxybenzoate + O2
3-carboxy-cis,cis-muconate
best substrate
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-
?
3,4-dihydroxybenzoate + O2
3-carboxy-cis,cis-muconate
100% activity
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-
?
3,4-dihydroxybenzoate + O2
3-carboxy-cis,cis-muconate
-
-
-
-
?
3,4-dihydroxybenzoate + O2
3-carboxy-cis,cis-muconate
100% activity
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-
?
3,4-dihydroxybenzoate + O2
3-carboxy-cis,cis-muconate
-
-
-
?
3,4-dihydroxybenzoate + O2
3-carboxy-cis,cis-muconate
best substrate
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-
?
3,4-dihydroxybenzoate + O2
3-carboxy-cis,cis-muconate
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no other substrate found
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?
3,4-dihydroxybenzoate + O2
?
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spectroscopic and electronic structure study of the enzyme-substrate complex
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-
?
3,4-dihydroxybenzoate + O2
?
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spectroscopic and electronic structure study of the enzyme-substrate complex
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?
3,4-dihydroxyhydrocinnamic acid + O2
?
29.5% of the activity with protocatechuate for the free enzyme, 95.0% for the calcium alginate-immobilized enzyme, and 41.1% for the glyoxyl agarose-immobilized enzyme
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-
?
3,4-dihydroxyhydrocinnamic acid + O2
?
29.54% ctivity compared to 3,4-dihydroxybenzoate
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-
?
3,4-dihydroxyhydrocinnamic acid + O2
?
29.54% ctivity compared to 3,4-dihydroxybenzoate
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-
?
3,4-dihydroxyhydrocinnamic acid + O2
?
29.5% of the activity with protocatechuate for the free enzyme, 95.0% for the calcium alginate-immobilized enzyme, and 41.1% for the glyoxyl agarose-immobilized enzyme
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-
?
3,4-dihydroxymandelic acid + O2
?
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protocatechuate 3,4-dioxygenase II
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-
?
3,4-dihydroxymandelic acid + O2
?
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protocatechuate 3,4-dioxygenase II
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-
?
3,4-dihydroxymandelic acid + O2
?
Nocardia erythropolis
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at 5.4% the rate of protocatechuic acid oxidation
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-
?
3,4-dihydroxyphenylacetate + O2
?
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W153V protocatechuate 3,4-dioxygenase I mutant enzyme: 13% activity compared to protocatechuate, wild-type protocatechuate 3,4-dioxygenase I: 2% activity compared to protocatechuate
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-
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3,5-dihydroxybenzoate + O2
?
44.3% of the activity with protocatechuate for the free enzyme, 113.3% for the calcium alginate-immobilized enzyme, and 42.9% for the glyoxyl agarose-immobilized enzyme
-
-
?
3,5-dihydroxybenzoate + O2
?
44.33% activity compared to 3,4-dihydroxybenzoate
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-
?
3-methylcatechol + O2
2-methylmuconate
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at 5% the rate of protocatechuic acid oxidation
-
?
3-methylcatechol + O2
2-methylmuconate
Nocardia erythropolis
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at 14.4% the rate of protocatechuic acid oxidation
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?
3-methylcatechol + O2
2-methylmuconate
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at 0.4% the rate of protocatechuic acid oxidation
-
?
4-methylcatechol + O2
3-methyl-cis,cis-muconate
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at very low rates
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?
4-methylcatechol + O2
3-methyl-cis,cis-muconate
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at very low rates
-
?
4-methylcatechol + O2
3-methyl-cis,cis-muconate
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at very low rates
-
?
4-methylcatechol + O2
3-methyl-cis,cis-muconate
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at very low rates
-
?
4-methylcatechol + O2
3-methyl-cis,cis-muconate
Nocardia erythropolis
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at 3.1% the rate of protocatechuic acid oxidation
-
?
4-sulfocatechol + O2
3-sulfomuconate
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catalyzed by protocatechuate 3,4-dioxgenase type II only
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?
4-sulfocatechol + O2
3-sulfomuconate
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catalyzed by protocatechuate 3,4-dioxgenase type II only
-
?
4-sulfocatechol + O2
3-sulfomuconate
-
catalyzed by protocatechuate 3,4-dioxgenase type II only
-
?
caffeic acid + O2
?
34.2% of the activity with protocatechuate for the free enzyme, 150% for the calcium alginate-immobilized enzyme, and 97% for the glyoxyl agarose-immobilized enzyme
-
-
?
caffeic acid + O2
?
34.21% activity compared to 3,4-dihydroxybenzoate
-
-
?
caffeic acid + O2
?
34.21% activity compared to 3,4-dihydroxybenzoate
-
-
?
caffeic acid + O2
?
34.2% of the activity with protocatechuate for the free enzyme, 150% for the calcium alginate-immobilized enzyme, and 97% for the glyoxyl agarose-immobilized enzyme
-
-
?
catechol + O2
cis,cis-muconate
-
reaction of EC 1.13.11.1
-
-
?
catechol + O2
cis,cis-muconate
-
reaction of EC 1.13.11.1
-
-
?
catechol + O2
muconate
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at 3% the rate of protocatechuic acid oxidation
-
?
catechol + O2
muconate
Nocardia erythropolis
-
at 33.8% the rate of protocatechuic acid oxidation
-
?
catechol + O2
muconate
-
at 0.4% the rate of protocatechuic acid oxidation
-
?
catechol + O2
muconate
-
at 0.4% the rate of protocatechuic acid oxidation
-
?
pyrogallol + O2
?
-
at 10% the rate of protocatechuic acid oxidation
-
-
?
pyrogallol + O2
?
Nocardia erythropolis
-
at 36% the rate of protocatechuic acid oxidation
-
-
?
pyrogallol + O2
?
-
at 0.4% the rate of protocatechuic acid oxidation
-
-
?
pyrogallol + O2
?
-
at 0.4% the rate of protocatechuic acid oxidation
-
-
?
trans-3,4-dihydroxycinnamate + O2
?
-
W153V protocatechuate 3,4-dioxygenase I mutant enzyme: 35% activity compared to protocatechuate, wild-type protocatechuate 3,4-dioxygenase I: 2% activity compared to protocatechuate
-
-
-
additional information
?
-
benzoate induces expression of protocatechuate 3,4-dioxygenase
-
-
-
additional information
?
-
benzoate induces expression of protocatechuate 3,4-dioxygenase
-
-
-
additional information
?
-
benzoate induces expression of protocatechuate 3,4-dioxygenase
-
-
-
additional information
?
-
benzoate induces expression of protocatechuate 3,4-dioxygenase
-
-
-
additional information
?
-
-
enzyme involved in last ring fission in naphtalene degradation, intermediates confirm proceeding through protocatechuic acid via ortho-claevage pathway
-
-
-
additional information
?
-
-
the enzyme is active on a wide range of phenyl compounds, in contrast to the high specificity of similar enzymes from other sources
-
-
-
additional information
?
-
-
the enzyme is active on a wide range of phenyl compounds, in contrast to the high specificity of similar enzymes from other sources
-
-
-
additional information
?
-
-
strain KB2 degrades 13 mM 3,4-dihydroxybenzoate, 10 mM benzoic acid and 12 mM phenol within 24 h of incubation
-
-
-
additional information
?
-
protocatechuate 3,4-dioxygenase from KB2 strain shows activity against various dihydroxybenzoic acids, but highest activity with primary substrate protocatechuate. Activity of the enzyme immobilized on calcium alginate increases particularly towards 2,5-dihydroxybenzoate, caffeic acid, 2,3-dihydroxybenzoate, and 3,5-dihydroxybenzoate, overview
-
-
-
additional information
?
-
-
protocatechuate 3,4-dioxygenase from KB2 strain shows activity against various dihydroxybenzoic acids, but highest activity with primary substrate protocatechuate. Activity of the enzyme immobilized on calcium alginate increases particularly towards 2,5-dihydroxybenzoate, caffeic acid, 2,3-dihydroxybenzoate, and 3,5-dihydroxybenzoate, overview
-
-
-
additional information
?
-
-
strain KB2 degrades 13 mM 3,4-dihydroxybenzoate, 10 mM benzoic acid and 12 mM phenol within 24 h of incubation
-
-
-
additional information
?
-
protocatechuate 3,4-dioxygenase from KB2 strain shows activity against various dihydroxybenzoic acids, but highest activity with primary substrate protocatechuate. Activity of the enzyme immobilized on calcium alginate increases particularly towards 2,5-dihydroxybenzoate, caffeic acid, 2,3-dihydroxybenzoate, and 3,5-dihydroxybenzoate, overview
-
-
-
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NATURAL SUBSTRATE
NATURAL PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
3,4-dihydroxybenzoate + O2
3-carboxy-cis,cis-muconate
-
-
-
-
?
3,4-dihydroxybenzoate + O2
3-carboxy-cis,cis-muconate
-
-
-
-
?
3,4-dihydroxybenzoate + O2
3-carboxy-cis,cis-muconate
-
-
-
?
3,4-dihydroxybenzoate + O2
3-carboxy-cis,cis-muconate
-
-
-
?
3,4-dihydroxybenzoate + O2
3-carboxy-cis,cis-muconate
-
-
-
?
3,4-dihydroxybenzoate + O2
3-carboxy-cis,cis-muconate
100% activity
-
-
?
3,4-dihydroxybenzoate + O2
3-carboxy-cis,cis-muconate
-
-
-
-
?
3,4-dihydroxybenzoate + O2
3-carboxy-cis,cis-muconate
100% activity
-
-
?
3,4-dihydroxybenzoate + O2
3-carboxy-cis,cis-muconate
-
-
-
?
additional information
?
-
benzoate induces expression of protocatechuate 3,4-dioxygenase
-
-
-
additional information
?
-
benzoate induces expression of protocatechuate 3,4-dioxygenase
-
-
-
additional information
?
-
benzoate induces expression of protocatechuate 3,4-dioxygenase
-
-
-
additional information
?
-
benzoate induces expression of protocatechuate 3,4-dioxygenase
-
-
-
additional information
?
-
-
enzyme involved in last ring fission in naphtalene degradation, intermediates confirm proceeding through protocatechuic acid via ortho-claevage pathway
-
-
-
additional information
?
-
-
the enzyme is active on a wide range of phenyl compounds, in contrast to the high specificity of similar enzymes from other sources
-
-
-
additional information
?
-
-
the enzyme is active on a wide range of phenyl compounds, in contrast to the high specificity of similar enzymes from other sources
-
-
-
additional information
?
-
-
strain KB2 degrades 13 mM 3,4-dihydroxybenzoate, 10 mM benzoic acid and 12 mM phenol within 24 h of incubation
-
-
-
additional information
?
-
-
strain KB2 degrades 13 mM 3,4-dihydroxybenzoate, 10 mM benzoic acid and 12 mM phenol within 24 h of incubation
-
-
-
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
2,2'-dipyridyl
activates the enzyme immobilized on calcium alginate, but inhibits the free enzyme and the enzyme immobilized on glyoxyl agarose, at 0.3 mM
Al3+
activates the free enzyme and enzyme immobilized on calcium alginate
Cu2+
-
increases enzyme activity for both cell-free and immobilized extracts by 8% and 44%, respectively
EDTA
activates the enzyme immobilized on calcium alginate, but inhibits the free enzyme and the enzyme immobilized on glyoxyl agarose, at 0.3 mM
Fe
Fe3+
additional information
Fe
-
external addition of FeSO4 is absolutely essential, other metal ions cannot replace Fe2+
Fe3+
-
increases enzyme activity for both cell-free and immobilized extracts by 16% and 99%, respectively
Fe3+
-
presence of a alphabetaFe3+ protomer in a variety of stoichiometries
Fe3+
activates the enzyme immobilized on calcium alginate
additional information
protective effect of immobilization on calcium alginate on enzyme activity is observed in the presence of Cd2+, Al3+, and Fe3+ while in the presence of Mn2+ and Zn2+ the inhibitory effect of metals on enzyme activity increases
additional information
-
protective effect of immobilization on calcium alginate on enzyme activity is observed in the presence of Cd2+, Al3+, and Fe3+ while in the presence of Mn2+ and Zn2+ the inhibitory effect of metals on enzyme activity increases
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INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
(2E)-3-(3,4-dimethoxyphenyl)-N-hydroxyprop-2-enamide
over 90% inhibition at 0.2 mM
(2E)-N-hydroxy-3-(4-methoxyphenyl)prop-2-enamide
over 90% inhibition at 0.2 mM
1,10-phenanthroline
-
0.005 mM, approx. 50% inhibition after 30 min, complete inhibition after 60 min, complete restoration of inactivated enzyme by addition of excess ferric EDTA complex
2-(2H-1,3-benzodioxol-5-yl)-N-[(4-fluorophenyl)methyl]-N-hydroxyacetamide
60% inhibition at 0.2 mM
3-(3,4-dimethoxyphenyl)-N-hydroxy-N-octylpropanamide
57% inhibition at 0.2 mM
3-(3,4-dimethoxyphenyl)-N-hydroxypropanamide
60% inhibition at 0.2 mM
2,2'-dipyridyl
activates the enzyme immobilized on calcium alginate, but inhibits the free enzyme and the enzyme immobilized on glyoxyl agarose, at 0.3 mM
2-Hydroxyisonicotinic acid N-oxide
-
0.000025 mM, 50% inactivation after 10 min, most potent inhibitor
Al3+
slight inhibition of the glyoxyl agarose-immobilized enzyme
butanol
inhibits the activity of the enzyme immobilized on glyoxyl agarose by 23.5% at 0.3 mM
EDTA
activates the enzyme immobilized on calcium alginate, but inhibits the free enzyme and the enzyme immobilized on glyoxyl agarose, at 0.3 mM
ethanol
inhibits the activity of the enzyme immobilized on glyoxyl agarose by 50% at 0.3 mM
Fe3+
inhibits the free enzyme and the enzyme immobilized on glyoxyl agarose
methanol
inhibits the activity of the enzyme immobilized on calcium alginate by 28% at 0.3 mM
Propanol
inhibits the activity of the enzyme immobilized on glyoxyl agarose by 23% at 0.3 mM
additional information
-
not inhibited by iodoacetamide, 5,5'-dithiobis(2-nitrobenzoate), mercaptoethanol, dithiothreitol, glutathione, o-phenanthroline and H2O2
-
additional information
-
not inhibited by arsenate, iodoacetate, semicarbazide, N-phenylmaleimide, fluoride or cyanide
-
additional information
poor inhibition by N-hydroxy-3-(4-methoxyphenyl)propanamide at 0.2 mM
-
additional information
-
poor inhibition by N-hydroxy-3-(4-methoxyphenyl)propanamide at 0.2 mM
-
additional information
protective effect of immobilization on calcium alginate on enzyme activity is observed in the presence of Cd2+, Al3+, and Fe3+ while in the presence of Mn2+ and Zn2+ the inhibitory effect of metals on enzyme activity increases
-
additional information
-
protective effect of immobilization on calcium alginate on enzyme activity is observed in the presence of Cd2+, Al3+, and Fe3+ while in the presence of Mn2+ and Zn2+ the inhibitory effect of metals on enzyme activity increases
-
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ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
butanol
activates the free enzyme and enzyme immobilized on calcium alginate by 20% and 9% at 0.3 mM, respectively
ethanol
activates the free enzyme and enzyme immobilized on calcium alginate by 42% and 56% at 0.3 mM, respectively
Propanol
activates the free enzyme by 30% at 0.3 mM
additional information
propanol has no effect on the activity of enzyme immobilized on calcium alginate at 0.3 mM
-
additional information
-
propanol has no effect on the activity of enzyme immobilized on calcium alginate at 0.3 mM
-
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KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.06534
3,4-dihydroxybenzoate
free enzyme, at pH 7.4 and 37°C
0.095
3,4-dihydroxybenzoate
-
multi-walled carbon nanotube-immobilized enzyme, at pH 10.0 and 60°C
0.15929
3,4-dihydroxybenzoate
Fe3O4 nanoparticles-immobilized enzyme, at pH 7.4 and 37°C
additional information
additional information
-
measurement of Km at different pH values
-
additional information
additional information
-
rates (s-1) and dissocation constants (microM) for protocatechuate: Y408H: k+2 = 2.8, k-2 = 1.9, k+2 + k-2 = 4.7, K1 = 2600, Kd = 1100, Y408F: k+2 = 0.39, k-2 = 0.46, k+2 + k-2 = 0.85, K1 = 84, Kd = 45, Y408C: k+2 + k-2 = 0.67; substrate dissociation constant for protocatechuate: wild type = 2.5 microM, Y408H = 39 micro M, Y408C = 57 microM
-
additional information
additional information
-
the enzyme exhibits Michaelis-Menten kinetics for all substrates tested
-
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TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
4.3
3,4-dihydroxybenzoate
-
multi-walled carbon nanotube-immobilized enzyme, at pH 10.0 and 60°C
10.25
3,4-dihydroxybenzoate
free enzyme, at pH 7.4 and 37°C
15.97
3,4-dihydroxybenzoate
Fe3O4 nanoparticles-immobilized enzyme, at pH 7.4 and 37°C
0.007
protocatechuate
-
for the mutant enzyme Y408H, at 25°C in 50 mM Tris and 2 mM beta-mercaptoethanol, pH 8.5 under saturating protocatechuate and O2 conditions
0.01
protocatechuate
-
for the mutant enzyme Y408E, at 25°C in 50 mM Tris and 2 mM beta-mercaptoethanol, pH 8.5 under saturating protocatechuate and O2 conditions
0.011
protocatechuate
-
for the mutant enzyme Y408F, at 25°C in 50 mM Tris and 2 mM beta-mercaptoethanol, pH 8.5 under saturating protocatechuate and O2 conditions
0.13
protocatechuate
-
for the mutant enzyme Y408C, at 25°C in 50 mM Tris and 2 mM beta-mercaptoethanol, pH 8.5 under saturating protocatechuate and O2 conditions
100
protocatechuate
-
for the wild-type enzyme, at 25°C in 50 mM Tris and 2 mM beta-mercaptoethanol, pH 8.5 under saturating protocatechuate and O2 conditions
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kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
45.1
3,4-dihydroxybenzoate
-
multi-walled carbon nanotube-immobilized enzyme, at pH 10.0 and 60°C
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Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.0068
2-Hydroxyisonicotinic acid N-oxide
-
apparent Ki, irreversible binding
3.9
2-Hydroxypyridine N-oxide
-
apparent Ki, inhibition is not freely reversible
additional information
additional information
-
rates (s-1) and dissociation constants (microM) for 4-hysroxybenzoate: wild type: k+2 = 42, k-2 = 14, k+2 + k-2 = 56, K1 = 700, Kd = 170, Y408H: k+2 + k-2 = 2.3, Y408C: k+2 + k-2 = 0.7, Y408F: k+2 + k-2 = 0.9; substrate dissociation constant for 3-hydroxybenzoate: wild type = 3500 microM, Y408H = 180 microM, Y408E = 210 microM, Y408C = 2500 microM, Y408F = 1800 microM; substrate dissociation constant for 4-hydroxybenzoate: wild type = 300 microM, Y408H = 6.3 microM, Y408C = 51 microM, Y408F = 12 microM
-
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IC50 VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.02
(2E)-3-(3,4-dimethoxyphenyl)-N-hydroxyprop-2-enamide
Rhodococcus jostii
pH 7.5, 37°C
0.015
(2E)-N-hydroxy-3-(4-methoxyphenyl)prop-2-enamide
Rhodococcus jostii
pH 7.5, 37°C
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SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
0.113
-
activity in E. coli cell extracts expressing the recombinant enzyme
16.7
-
type II protocatechuate 3,4-dioxygenase, oxidation of 4-sulfocatechol
2.98
-
inductor 3,4-dihydroxybenzoic acid, cell-free extract from Stenotrophomonas maltophilia
5.2
-
type II protocatechuate 3,4-dioxygenase, oxidation of 4-sulfocatechol
6.37
-
inductor 4-hydroxybenzoic acid, cell-free extract from Stenotrophomonas maltophilia
8.7
-
inductor vanillic acid, cell-free extract from Stenotrophomonas maltophilia
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pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
7.5
8.5
9.2
8.5
-
maximum activity for the wild type enzyme and the mutants enymes Y408C, Y408F, Y408E and Y408H
9.2
free enzyme and glyoxyl agarose-immobilized enzyme
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pH RANGE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
2.2 - 13
glyoxyl agarose-immobilized enzyme, profile overview
4 - 14
calcium alginate-immobilized enzyme, profile overview
5.5 - 8.5
the enzyme shows more than 50% activity between pH 5.5 and 8.5
6 - 9
-
pH 6.0: about 40% of maximal activity, pH 9.0: about 65% of maximal activity
7 - 11
the enzyme shows more than 50% activity between pH 7.0 and 11.0 and loses 98.3% of its original enzymatic activity at pH 4.0 and about 95.4% at pH 12.0
additional information
-
activity increases steadily up to pH 9.0, above pH 9.0 the enzyme begins to undergo spontaneous oxidation
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TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
35
37
40
45 - 50
55
60
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TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
20 - 70
the enzyme shows more than 50% activity between 20 and 70°C
4 - 60
free and immobilized enzyme, profile overview
50 - 70
-
50°C: about 55% of maximal activity, 70°C: about 60% of maximal activity
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ORGANISM
COMMENTARY
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
enzyme is induced in cells grown on protocatechuic acid or analogues
-
-
brenda
Pseudomonas mendocina P2d, an array of colors are formed when Pseudomonas mendocia P2d cells grow in sodium or tyrosine, which is ascribed to the presence of multiple enzyms: catechol-1,2-doxygenase, catechol-2,3-dioxygenase, protocatechuate-3,4-dioxygenase, and tyrosinase
-
-
brenda
isolates GAI-16, S25com04 and IC4, enzyme activity is induced by growth on p-hydroxybenzoate
-
-
brenda
protocatechuate 3,4-dioxygenase alpha subunit
SwissProt
brenda
protocatechuate 3,4-dioxygenase beta subunit
SwissProt
brenda
protocatechuate 3,4-dioxygenase alpha subunit
SwissProt
brenda
protocatechuate 3,4-dioxygenase beta subunit
SwissProt
brenda
cells cultivated with protocatechuate, p-cresol, vanillate and 4-hydroybenzoate as sole carbon and energy surce for growth, exhibite protocatechuate 3,4-dioxygenase activities
-
-
brenda
thermophilic isolate G27 from high-temperature oilfield in Lithuania
-
-
brenda
American Type Culture Collection (ATCC) 23975, previously classified as Pseudomonas aeruginosa
-
-
brenda
examining a distorted trigonal-bipyramidal geometry observed for the non-heme iron center in protocatechuate 3,4-dioxygenase by utilizing a sterically hindered iron salen complex
-
-
brenda
previously classified as Pseudomonas aeruginosa strain ATCC 23975
-
-
brenda
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SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
SOURCE
-
monocyclic hydrocarbons, 4-hydroxybenzoic acid, protocatechuic acid and vanillic acid
brenda
-
monocyclic hydrocarbons, 4-hydroxybenzoic acid, protocatechuic acid and vanillic acid
-
brenda
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LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
-
localized in 100000 g supernatant, completely absent in mitochondrial, microsomal and chloroplast fraction
brenda
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GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
metabolism
additional information
metabolism
-
protocatechuate 3,4-dioxygenase is the key enzyme of protocatechuate breakdown
metabolism
-
PCA 3,4-dioxygenase is essential for vanillate, 4-hydroxybenzoate, and protocatechuate assimilation
metabolism
-
the enzyme is involved in the degradation of the environmental pollutant 3-chlorobenzoate
metabolism
-
protocatechuate 3,4-dioxygenase is the key enzyme of protocatechuate breakdown
-
metabolism
-
protocatechuate 3,4-dioxygenase is the key enzyme of protocatechuate breakdown
-
metabolism
-
the enzyme is involved in the degradation of the environmental pollutant 3-chlorobenzoate
-
additional information
enzyme immobilized on calcium alginate gel binds to the carrier by electrostatic interaction while glyoxyl agarose is linked with the enzyme by covalent bonds. Since positively charged amino acids are localized at the entrance of crevasse, the electrostatic interaction plays the key role in modulation of enzyme activity. Significant increase of enzyme activity after its immobilization in calcium alginate is probably caused by strong electrostatic interactions between positively charged amino acid residues of the enzyme and negatively charged groups of alginate. High activity of immobilized protocatechuate 3,4-dioxygenase towards 2,3-dihydroxybenzoate, 2,5-dihydroxybenzoate, and 3,5-dihydroxybenzoate can be connected with modification of its catalytic mechanism as only 2,3-dihydroxybenzoate is in configuration typical for protocatechuate 3,4-dioxygenase's substrate
additional information
-
enzyme immobilized on calcium alginate gel binds to the carrier by electrostatic interaction while glyoxyl agarose is linked with the enzyme by covalent bonds. Since positively charged amino acids are localized at the entrance of crevasse, the electrostatic interaction plays the key role in modulation of enzyme activity. Significant increase of enzyme activity after its immobilization in calcium alginate is probably caused by strong electrostatic interactions between positively charged amino acid residues of the enzyme and negatively charged groups of alginate. High activity of immobilized protocatechuate 3,4-dioxygenase towards 2,3-dihydroxybenzoate, 2,5-dihydroxybenzoate, and 3,5-dihydroxybenzoate can be connected with modification of its catalytic mechanism as only 2,3-dihydroxybenzoate is in configuration typical for protocatechuate 3,4-dioxygenase's substrate
additional information
-
enzyme immobilized on calcium alginate gel binds to the carrier by electrostatic interaction while glyoxyl agarose is linked with the enzyme by covalent bonds. Since positively charged amino acids are localized at the entrance of crevasse, the electrostatic interaction plays the key role in modulation of enzyme activity. Significant increase of enzyme activity after its immobilization in calcium alginate is probably caused by strong electrostatic interactions between positively charged amino acid residues of the enzyme and negatively charged groups of alginate. High activity of immobilized protocatechuate 3,4-dioxygenase towards 2,3-dihydroxybenzoate, 2,5-dihydroxybenzoate, and 3,5-dihydroxybenzoate can be connected with modification of its catalytic mechanism as only 2,3-dihydroxybenzoate is in configuration typical for protocatechuate 3,4-dioxygenase's substrate
-
Show AA Sequence and Transmembrane Helices (4298 entries)
Please use the AA Sequence and Transmembrane Helices Search for a specific query.
Please use the AA Sequence and Transmembrane Helices Search for a specific query.
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PDB
SCOP
CATH
UNIPROT
ORGANISM
Acinetobacter baylyi (strain ATCC 33305 / BD413 / ADP1)
Acinetobacter baylyi (strain ATCC 33305 / BD413 / ADP1)
Acinetobacter baylyi (strain ATCC 33305 / BD413 / ADP1)
Acinetobacter baylyi (strain ATCC 33305 / BD413 / ADP1)
Acinetobacter baylyi (strain ATCC 33305 / BD413 / ADP1)
Acinetobacter baylyi (strain ATCC 33305 / BD413 / ADP1)
Acinetobacter baylyi (strain ATCC 33305 / BD413 / ADP1)
Acinetobacter baylyi (strain ATCC 33305 / BD413 / ADP1)
Acinetobacter baylyi (strain ATCC 33305 / BD413 / ADP1)
Acinetobacter baylyi (strain ATCC 33305 / BD413 / ADP1)
Acinetobacter baylyi (strain ATCC 33305 / BD413 / ADP1)
Acinetobacter baylyi (strain ATCC 33305 / BD413 / ADP1)
Acinetobacter baylyi (strain ATCC 33305 / BD413 / ADP1)
Acinetobacter baylyi (strain ATCC 33305 / BD413 / ADP1)
Acinetobacter baylyi (strain ATCC 33305 / BD413 / ADP1)
Acinetobacter baylyi (strain ATCC 33305 / BD413 / ADP1)
Acinetobacter baylyi (strain ATCC 33305 / BD413 / ADP1)
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MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
150000
21900
-
x * 21900 + x * 26700, deduced from nucleotide sequences of putative alpha and beta subunits
22280
-
alpha,beta, 2 * 22280 + 2 * 26630, holoenzyme probably contains 6 alpha2,beta2 tetramers, amino acid sequence
23000
26500
26630
-
alpha,beta, 2 * 22280 + 2 * 26630, holoenzyme probably contains 6 alpha2,beta2 tetramers, amino acid sequence
26700
-
x * 21900 + x * 26700, deduced from nucleotide sequences of putative alpha and beta subunits
29000
34300
-
trimer of dimers, (alphabeta)3, 3 * 29000, alpha subunit, + 3 * 34300, beta-subunit, SDS-PAGE
480000
29000
-
trimer of dimers, (alphabeta)3, 3 * 29000, alpha subunit, + 3 * 34300, beta-subunit, SDS-PAGE
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SUBUNIT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
decamer
heterodimer
hexamer
octamer
tetramer
-
alpha,beta, 2 * 22280 + 2 * 26630, holoenzyme probably contains 6 alpha2,beta2 tetramers, amino acid sequence
additional information
?
-
x * 21900 + x * 26700, deduced from nucleotide sequences of putative alpha and beta subunits
heterodimer
1 * 22400 + 1 * 26800, calculated from amino acid sequence
heterodimer
-
1 * 22400 + 1 * 26800, calculated from amino acid sequence
-
hexamer
-
trimer of dimers, (alphabeta)3, 3 * 29000, alpha subunit, + 3 * 34300, beta-subunit, SDS-PAGE
hexamer
-
trimer of dimers, (alphabeta)3, 3 * 29000, alpha subunit, + 3 * 34300, beta-subunit, SDS-PAGE
-
additional information
-
amino acid sequence of beta-subunit, 238 amino acids
additional information
-
amino acid sequence of alpha-subunit, 203 amino acids
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POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
no glycoprotein
-
carbohydrate contributes less than 0.2% to the mass of the holoenzyme
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CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
recombinant enzyme, hanging drop method with 1.8 M ammonium sulfate, 50 mM Tris-HCl pH 7.0 in the reservoir and 10 mg/ml enzyme in the drop at 18°C, crystals grow as perfect dodecahedrons, crystal structure with 2.2 A resolution
-
microdialysis of approx. 0.05 ml of enzyme solution, 80 mg/ml against 10 ml volumes of buffered solutions, large single crystals
-
vapor equilibration at room temperature from solutions containing potassium phosphate pH 7.5, 1.08-1.2 M reservoir concentrations, initial enzyme concentration 6.7-10.5 mg/ml
-
addition of 0.1 ml 1 M 2-mercaptoethanol to 10 ml enzyme solution in 50 mM Tris buffer pH 8.5, rhombic crystals appear after 1 week in a refrigerator, crystallization is also achieved by adding 1 mM iodoacetamide instead of mercaptoethanol or by dialyzing the enzyme against cold, destilled water for 2 days
-
free interface diffusion cell, equal volumes of concentrated enzyme and ammonium sulfate solutions ranging from 37-47% saturation, 4°C, 2.5 A resolution
-
vapor diffusion in hanging drops, crystal structure at 2.1-2.2 A resolution
-
vapor diffusion with ammonium sulfate as the precipitant at 4°C, recombinant wild-type and Y447H mutant enzyme
-
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PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
DELTA319-322
-
turnover-number is 4.14fold lower than that of the wild-type enzyme, the Km-value for 3,4-dihydroxybenzoate is 2.1fold lower than that of the wild-type enzyme
R133H
R457S
-
turnover-number is 1333fold lower than that of the wild-type enzyme, the Km-value for 3,4-dihydroxybenzoate is 2.2fold lower than that of the wild-type enzyme
R133H
-
gain of function mutation confers catechol 1,2-dioxygenase activity
-
R142K
-
like wild-type no acticity of mutated protocatechuate 3,4-dioxygenase I with 4-sulfocatechol
R142K/W153V
-
protocatechuate 3,4-dioxygenase I gain of function mutation, mutant enzyme oxidizes 4-sulfocatechol
R153V
-
protocatechuate 3,4-dioxygenase I gain of function mutation, mutant enzyme oxidizes 4-sulfocatechol
Y408C
Y408E
Y408F
-
iron is not tightly bound, the Y408F mutant does not reconstitute above half-occupancy and loses color during crystallization attempts. Inhibitors like 4-hydroybenzoate and 3-hydroybenzoate bind more tighly to the mutant enzyme, whereas the substrate protocatechuate binds less tightly.
Y408H
Y447H
additional information
R133H
-
gain of function mutation confers catechol 1,2-dioxygenase activity
R133H
-
turnover-number is fold 500lower than that of the wild-type enzyme, the Km-value for 3,4-dihydroxybenzoate is 1.8fold higher than that of the wild-type enzyme
Y408C
-
turnover-number is 523fold lower than that of the wild-type enzyme
Y408C
-
iron is tightly bound. The structure reveals no significant mutation-related changes except in the immediate vicinity of the altered amino acid (rmsd over all atoms = 0.2-0.3 A). The new amino acid does not coordinate to the iron, because the side chain is shorter than that of Tyr. In contrast to the wild-type enzyme, Tyr447 remains bound to the iron, as a result, a monodentate substrate complex is formed between the iron and protocatechuate 04. Protocatechuate does not shift into a chelated orientation. Inhibitors like 4-hydroybenzoate and 3-hydroybenzoate bind more tighly to the mutant enzyme, whereas the substrate protocatechuate binds less tightly.
Y408E
-
turnover-number is 6800fold lower than that of the wild-type enzyme
Y408E
-
iron is tightly bound. The structure reveals no significant mutation-related changes except in the immediate vicinity of the altered amino acid (rmsd over all atoms = 0.2-0.3 A). The new amino acid does not coordinate to the iron, because the side chain is shorter than that of Tyr. In contrast to the wild-type enzyme, Tyr447 remains bound to the iron, as a result, a monodentate substrate complex is formed between the iron and protocatechuate 04. Protocatechuate does not shift into a chelated orientation.
Y408H
-
turnover-number is 9714fold lower than that of the wild-type enzyme, the Km-value for 3,4-dihydroxybenzoate is 10fold lower than that of the wild-type enzyme
Y408H
-
iron is tightly bound. The structure reveals no significant mutation-related changes except in the immediate vicinity of the altered amino acid (rmsd over all atoms = 0.2-0.3 A). The new amino acid does not coordinate to the iron, because the side chain is shorter than that of Tyr. In contrast to the wild-type enzyme, Tyr447 remains bound to the iron, as a result, a monodentate substrate complex is formed between the iron and protocatechuate 04. Protocatechuate does not shift into a chelated orientation. Inhibitors like 4-hydroybenzoate and 3-hydroybenzoate bind more tighly to the mutant enzyme, whereas the substrate protocatechuate binds less tightly.
Y447H
-
turnover-number is 567fold lower than that of the wild-type enzyme
additional information
-
mutants are constructed so that their pcaG genes contained variations in repeat sequence capable of producing a selectable phenotype following a specific deletion. Deletion frequencies of the various mutations is determined and compared with repair frequencies of three different single-base mutations.
additional information
-
immobilization of the enzyme, the immobilized extract exhibited higher enzyme activity than the cell-free extract in the presence of trace elements and cations
additional information
immobilization of the enzyme in alginate gel shifts its optimum pH towards high-alkaline pH while immobilization of the enzyme on glyoxyl agarose does not influence pH-profile of the enzyme. Protocatechuate 3,4-dioygenase immobilized in calcium alginate shows increased activity towards 2,5-dihydroxybenzoate, caffeic acid, 2,3-dihydroxybenzoate, and 3,5-dihydroxybenzoate. Slightly lower activity of the enzyme is observed after its immobilization on glyoxyl agarose. Entrapment of the enzyme in alginate gel protects it against chelators and aliphatic alcohols while its immobilization on glyoxyl agarose enhanced enzyme resistance to inactivation by metal ions. Immobilization of dioxygenase in calcium alginate or on glyoxyl agarose results in decrease in the optimum temperature by 5°C and10°C, respectively. Activity of the enzyme immobilized on calcium alginate increases particularly towards 2,5-dihydroxybenzoate, caffeic acid, 2,3-dihydroxybenzoate, and 3,5-dihydroxybenzoate
additional information
-
immobilization of the enzyme in alginate gel shifts its optimum pH towards high-alkaline pH while immobilization of the enzyme on glyoxyl agarose does not influence pH-profile of the enzyme. Protocatechuate 3,4-dioygenase immobilized in calcium alginate shows increased activity towards 2,5-dihydroxybenzoate, caffeic acid, 2,3-dihydroxybenzoate, and 3,5-dihydroxybenzoate. Slightly lower activity of the enzyme is observed after its immobilization on glyoxyl agarose. Entrapment of the enzyme in alginate gel protects it against chelators and aliphatic alcohols while its immobilization on glyoxyl agarose enhanced enzyme resistance to inactivation by metal ions. Immobilization of dioxygenase in calcium alginate or on glyoxyl agarose results in decrease in the optimum temperature by 5°C and10°C, respectively. Activity of the enzyme immobilized on calcium alginate increases particularly towards 2,5-dihydroxybenzoate, caffeic acid, 2,3-dihydroxybenzoate, and 3,5-dihydroxybenzoate
additional information
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immobilization of the enzyme in alginate gel shifts its optimum pH towards high-alkaline pH while immobilization of the enzyme on glyoxyl agarose does not influence pH-profile of the enzyme. Protocatechuate 3,4-dioygenase immobilized in calcium alginate shows increased activity towards 2,5-dihydroxybenzoate, caffeic acid, 2,3-dihydroxybenzoate, and 3,5-dihydroxybenzoate. Slightly lower activity of the enzyme is observed after its immobilization on glyoxyl agarose. Entrapment of the enzyme in alginate gel protects it against chelators and aliphatic alcohols while its immobilization on glyoxyl agarose enhanced enzyme resistance to inactivation by metal ions. Immobilization of dioxygenase in calcium alginate or on glyoxyl agarose results in decrease in the optimum temperature by 5°C and10°C, respectively. Activity of the enzyme immobilized on calcium alginate increases particularly towards 2,5-dihydroxybenzoate, caffeic acid, 2,3-dihydroxybenzoate, and 3,5-dihydroxybenzoate
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pH STABILITY
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
7.6 - 8.6
Nocardia erythropolis
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most stable
439502
8.5
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TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
50
60
60 - 70
after incubating for 330 min at 60°C, the free enzyme maintains approximately 17% of its activity, and its half-life is 180 min. The enzyme retains 75% activity after 4 h at 70°C and shows no activity after 4 h at 90°C
65
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soluble and immobilized enzyme, 15 min, complete loss of activity
90
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the free enzyme loses 82% of relative activity after 180 min of incubations at 90°C
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GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
enzyme immobilized on porous glass beads is stable over wide ranges of pH and temperature
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Fe3O4 nanoparticles-immobilized enzyme exhibits improved thermal stability compared to the free enzyme
immobilization: slight increase of thermal stability, soluble and immobilized enzyme exhibit substantial activity in 2 M urea
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multi-walled carbon nanotube-immobilized enzyme exhibits improved thermal and pH stabilities compared to the free enzyme
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no loss of activity over a period of 78 d under aerobic conditions at 15°C at pH 7.0, pH 8.0 or pH 9.0
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STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
4°C, stability of free and the immobilized protocatechuate 3,4-dioxygenase in phosphate buffer containing 50 mM, pH 7.0, for free enzyme or immobilized enzyme in calcium alginate, and 40mM, pH 8.0, for enzyme immobilized on glyoxyl agarose, immobilized enzyme shows 9.93% of its initial activity after 28 days of storage, while the free enzyme becomes inactive
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PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
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); enzymes from Pseudomonas florescens
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native enzyme 296.8fold by ammonium sulfate fractionation, anion exchange and hydrophobic interaction 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; 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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