Information on EC 1.13.11.3 - protocatechuate 3,4-dioxygenase

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The expected taxonomic range for this enzyme is: Bacteria, Eukaryota

EC NUMBER
COMMENTARY
1.13.11.3
-
RECOMMENDED NAME
GeneOntology No.
protocatechuate 3,4-dioxygenase
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
3,4-dihydroxybenzoate + O2 = 3-carboxy-cis,cis-muconate
show the reaction diagram
proposed mechanism based on Mssbauer, EPR and inhibition kinetic data
-
3,4-dihydroxybenzoate + O2 = 3-carboxy-cis,cis-muconate
show the reaction diagram
proposed reaction mechanism
-
3,4-dihydroxybenzoate + O2 = 3-carboxy-cis,cis-muconate
show the reaction diagram
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
show the reaction diagram
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.
-
3,4-dihydroxybenzoate + O2 = 3-carboxy-cis,cis-muconate
show the reaction diagram
-
-
-
-
REACTION TYPE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
oxidation
-
-
-
-
redox reaction
-
-
-
-
reduction
-
-
-
-
PATHWAY
KEGG Link
MetaCyc Link
Benzoate degradation
-
Metabolic pathways
-
Microbial metabolism in diverse environments
-
Polycyclic aromatic hydrocarbon degradation
-
protocatechuate degradation II (ortho-cleavage pathway)
-
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.
SYNONYMS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
3,4-PCDase
Geobacillus sp.
-
-
EC 1.13.1.3
-
-
formerly
-
oxygenase, protocatechuate 3,4-di-
-
-
-
-
P3,4O enzyme
Leifsonia sp.
-
-
PCA 3,4-dioxygenase
-
-
PcaG
Acinetobacter baylyi ADPU54, Acinetobacter baylyi Crc
-
-
-
PcaH
Acinetobacter baylyi ADPU54, Acinetobacter baylyi Crc
-
-
-
protocatchetuate 3,4-dioxygenase
Geobacillus sp.
-
-
protocatechuate 3,4-dioxygenase
-
-
protocatechuate 3,4-dioxygenase
-
-
protocatechuate 3,4-dioxygenase
Stenotrophomonas maltophilia KB2
-
-
-
protocatechuate 3,4-dioxygenase
Sulfitobacter sp.
-
-
protocatechuate oxygenase
-
-
-
-
protocatechuic 3,4-dioxygenase
-
-
-
-
protocatechuic 3,4-oxygenase
-
-
-
-
protocatechuic acid oxidase
-
-
-
-
EC 1.99.2.3
-
-
formerly
-
additional information
-
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
Acinetobacter baylyi ADP1
-
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
CAS REGISTRY NUMBER
COMMENTARY
9029-47-4
-
ORGANISM
COMMENTARY
LITERATURE
SEQUENCE CODE
SEQUENCE DB
SOURCE
strain ADP1
-
-
Manually annotated by BRENDA team
Acinetobacter baylyi ADP1
strain ADP1
-
-
Manually annotated by BRENDA team
Acinetobacter baylyi Crc
-
-
-
Manually annotated by BRENDA team
strain 80-1, derivative of ATCC 14987
-
-
Manually annotated by BRENDA team
Acinetobacter calcoaceticus 80-1
strain 80-1, derivative of ATCC 14987
-
-
Manually annotated by BRENDA team
strain K24, enzyme is induced by growth on p-hydroxybenzoate
-
-
Manually annotated by BRENDA team
strain K24, enzyme is induced by growth on p-hydroxybenzoate
-
-
Manually annotated by BRENDA team
strain ADP1
-
-
Manually annotated by BRENDA team
strain S2, protocatechuate 3,4-dioxygenase I and II
-
-
Manually annotated by BRENDA team
strain S2, protocatechuate 3,4-dioxygenase I and II
-
-
Manually annotated by BRENDA team
formerly classified as Pseudomonas cepacia
-
-
Manually annotated by BRENDA team
protocatechuate 3,4-dioxygenase alpha subunit; strain HS-2
SwissProt
Manually annotated by BRENDA team
protocatechuate 3,4-dioxygenase beta subunit; strain HS-2
SwissProt
Manually annotated by BRENDA team
protocatechuate 3,4-dioxygenase alpha subunit; strain HS-2
SwissProt
Manually annotated by BRENDA team
protocatechuate 3,4-dioxygenase beta subunit; strain HS-2
SwissProt
Manually annotated by BRENDA team
cells cultivated with protocatechuate, p-cresol, vanillate and 4-hydroybenzoate as sole carbon and energy surce for growth, exhibite protocatechuate 3,4-dioxygenase activities
-
-
Manually annotated by BRENDA team
Geobacillus sp.
strain AY946034
-
-
Manually annotated by BRENDA team
Geobacillus sp.
thermophilic isolate G27 from high-temperature oilfield in Lithuania
-
-
Manually annotated by BRENDA team
strain S1, protocatechuate 3,4-dioxygenase II
-
-
Manually annotated by BRENDA team
Hydrogenophaga palleronii S1
strain S1, protocatechuate 3,4-dioxygenase II
-
-
Manually annotated by BRENDA team
Leifsonia sp.
-
-
-
Manually annotated by BRENDA team
enzyme is induced in cells grown on protocatechuic acid or analogues
-
-
Manually annotated by BRENDA team
no activity in Pseudomonas fluorescens
-
-
-
Manually annotated by BRENDA team
Nocardia erythropolis
-
-
-
Manually annotated by BRENDA team
wood-degrading fungus
-
-
Manually annotated by BRENDA team
strain B-10, enzyme is induced in cells grown on 4-hydroxybenzoate
-
-
Manually annotated by BRENDA team
Pseudomonas aeruginosa B-10
strain B-10, enzyme is induced in cells grown on 4-hydroxybenzoate
-
-
Manually annotated by BRENDA team
strain A.3.12.
-
-
Manually annotated by BRENDA team
Pseudomonas fluorescens A.3.12.
strain A.3.12.
-
-
Manually annotated by BRENDA team
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
-
-
Manually annotated by BRENDA team
American Type Culture Collection (ATCC) 23975, previously classified as Pseudomonas aeruginosa
-
-
Manually annotated by BRENDA team
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
-
-
Manually annotated by BRENDA team
previously classified as Pseudomonas aeruginosa strain ATCC 23975
-
-
Manually annotated by BRENDA team
previously classified as Pseudomonas fluorescens PHK
-
-
Manually annotated by BRENDA team
reclassified from Pseudomonas aeruginosa
-
-
Manually annotated by BRENDA team
Roseobacter sp.
isolates GAI-16, S25com04 and IC4, enzyme activity is induced by growth on p-hydroxybenzoate
-
-
Manually annotated by BRENDA team
enzyme activity is induced by growth on p-hydroxybenzoate
-
-
Manually annotated by BRENDA team
Stenotrophomonas maltophilia KB2
-
-
-
Manually annotated by BRENDA team
Stenotrophomonas maltophilia KB2
strain KB2
-
-
Manually annotated by BRENDA team
Sulfitobacter sp.
strain EE-36, Y3F, and Y4I
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
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
Acinetobacter baylyi ADPU54, Acinetobacter baylyi Crc
-
protocatechuate 3,4-dioxygenase is the key enzyme of protocatechuate breakdown
-
SUBSTRATE
PRODUCT                      
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
3,4-dihydroxybenzoate + O2
3-carboxy-cis,cis-muconate
show the reaction diagram
-
-
-
?
3,4-dihydroxybenzoate + O2
3-carboxy-cis,cis-muconate
show the reaction diagram
-
-
-
?
3,4-dihydroxybenzoate + O2
3-carboxy-cis,cis-muconate
show the reaction diagram
-
-
-
?
3,4-dihydroxybenzoate + O2
3-carboxy-cis,cis-muconate
show the reaction diagram
-
-
-
?
3,4-dihydroxybenzoate + O2
3-carboxy-cis,cis-muconate
show the reaction diagram
-
-
-
?
3,4-dihydroxybenzoate + O2
3-carboxy-cis,cis-muconate
show the reaction diagram
-
-
-
-
3,4-dihydroxybenzoate + O2
3-carboxy-cis,cis-muconate
show the reaction diagram
-
-
-
-
3,4-dihydroxybenzoate + O2
3-carboxy-cis,cis-muconate
show the reaction diagram
-
-
-
-
3,4-dihydroxybenzoate + O2
3-carboxy-cis,cis-muconate
show the reaction diagram
-
-
-
-
3,4-dihydroxybenzoate + O2
3-carboxy-cis,cis-muconate
show the reaction diagram
-
-
-
?
3,4-dihydroxybenzoate + O2
3-carboxy-cis,cis-muconate
show the reaction diagram
-
-
-
?
3,4-dihydroxybenzoate + O2
3-carboxy-cis,cis-muconate
show the reaction diagram
-
-
-
?
3,4-dihydroxybenzoate + O2
3-carboxy-cis,cis-muconate
show the reaction diagram
-
-
-
?
3,4-dihydroxybenzoate + O2
3-carboxy-cis,cis-muconate
show the reaction diagram
-
-
-
?
3,4-dihydroxybenzoate + O2
3-carboxy-cis,cis-muconate
show the reaction diagram
-
-
-
?
3,4-dihydroxybenzoate + O2
3-carboxy-cis,cis-muconate
show the reaction diagram
-
-
-
-
?
3,4-dihydroxybenzoate + O2
3-carboxy-cis,cis-muconate
show the reaction diagram
-
-
-
?
3,4-dihydroxybenzoate + O2
3-carboxy-cis,cis-muconate
show the reaction diagram
-
-
-
-
?
3,4-dihydroxybenzoate + O2
3-carboxy-cis,cis-muconate
show the reaction diagram
-
-
-
?
3,4-dihydroxybenzoate + O2
3-carboxy-cis,cis-muconate
show the reaction diagram
-
-
-
-
3,4-dihydroxybenzoate + O2
3-carboxy-cis,cis-muconate
show the reaction diagram
-
-
-
?
3,4-dihydroxybenzoate + O2
3-carboxy-cis,cis-muconate
show the reaction diagram
-
-
-
-
?
3,4-dihydroxybenzoate + O2
3-carboxy-cis,cis-muconate
show the reaction diagram
-
-
-
-
3,4-dihydroxybenzoate + O2
3-carboxy-cis,cis-muconate
show the reaction diagram
-
-
-
?
3,4-dihydroxybenzoate + O2
3-carboxy-cis,cis-muconate
show the reaction diagram
-
-
-
-
3,4-dihydroxybenzoate + O2
3-carboxy-cis,cis-muconate
show the reaction diagram
-
-
-
?
3,4-dihydroxybenzoate + O2
3-carboxy-cis,cis-muconate
show the reaction diagram
-
-
-
-
?
3,4-dihydroxybenzoate + O2
3-carboxy-cis,cis-muconate
show the reaction diagram
-
-
-
?
3,4-dihydroxybenzoate + O2
3-carboxy-cis,cis-muconate
show the reaction diagram
-
-
-
-
?
3,4-dihydroxybenzoate + O2
3-carboxy-cis,cis-muconate
show the reaction diagram
-
-
-
?
3,4-dihydroxybenzoate + O2
3-carboxy-cis,cis-muconate
show the reaction diagram
-
-
-
-
3,4-dihydroxybenzoate + O2
3-carboxy-cis,cis-muconate
show the reaction diagram
-
-
-
-
3,4-dihydroxybenzoate + O2
3-carboxy-cis,cis-muconate
show the reaction diagram
-
-
-
?
3,4-dihydroxybenzoate + O2
3-carboxy-cis,cis-muconate
show the reaction diagram
-
-
-
-
3,4-dihydroxybenzoate + O2
3-carboxy-cis,cis-muconate
show the reaction diagram
-
-
-
?
3,4-dihydroxybenzoate + O2
3-carboxy-cis,cis-muconate
show the reaction diagram
Roseobacter sp., Sagittula stellata
-
-
-
?
3,4-dihydroxybenzoate + O2
3-carboxy-cis,cis-muconate
show the reaction diagram
Geobacillus sp.
-
-
-
-
?
3,4-dihydroxybenzoate + O2
3-carboxy-cis,cis-muconate
show the reaction diagram
Leifsonia sp.
-
-
-
-
?
3,4-dihydroxybenzoate + O2
3-carboxy-cis,cis-muconate
show the reaction diagram
-
-
-
-
?
3,4-dihydroxybenzoate + O2
3-carboxy-cis,cis-muconate
show the reaction diagram
-
very low activity with: 3,4-dihydroxyphenylacetic acid
-
?
3,4-dihydroxybenzoate + O2
3-carboxy-cis,cis-muconate
show the reaction diagram
-
specific for protocatechuate
-
?
3,4-dihydroxybenzoate + O2
3-carboxy-cis,cis-muconate
show the reaction diagram
-
methylene blue cannot replace O2 as electron acceptor
-
ir
3,4-dihydroxybenzoate + O2
3-carboxy-cis,cis-muconate
show the reaction diagram
-
very low activity with 2-chloro-protocatechuate
-
?
3,4-dihydroxybenzoate + O2
3-carboxy-cis,cis-muconate
show the reaction diagram
Nocardia erythropolis
-
enzyme is active on a wide range of o-dihydroxyphenyl compounds
-
?
3,4-dihydroxybenzoate + O2
3-carboxy-cis,cis-muconate
show the reaction diagram
-
very low activity with 3,4-dihydroxymandelic acid
-
?
3,4-dihydroxybenzoate + O2
3-carboxy-cis,cis-muconate
show the reaction diagram
-
absolute requirement for vicinal hydroxyl groups in the 3- and 4-position
-
ir
3,4-dihydroxybenzoate + O2
3-carboxy-cis,cis-muconate
show the reaction diagram
-
very low activity with 5-bromo-protocatechuate, very low activity with 5-chloro-protocatechuate
-
?
3,4-dihydroxybenzoate + O2
3-carboxy-cis,cis-muconate
show the reaction diagram
-
very low activity with 4'-methylcatechol
-
?
3,4-dihydroxybenzoate + O2
3-carboxy-cis,cis-muconate
show the reaction diagram
-
no other substrate found
-
ir
3,4-dihydroxybenzoate + O2
3-carboxy-cis,cis-muconate
show the reaction diagram
-
no other substrate found
-
?
3,4-dihydroxybenzoate + O2
3-carboxy-cis,cis-muconate
show the reaction diagram
-
i.e. protocatechuate
-
-
?
3,4-dihydroxybenzoate + O2
3-carboxy-cis,cis-muconate
show the reaction diagram
-
-
-
?
3,4-dihydroxybenzoate + O2
3-carboxy-cis,cis-muconate
show the reaction diagram
-
-
-
?
3,4-dihydroxybenzoate + O2
3-carboxy-cis,cis-muconate
show the reaction diagram
Acinetobacter baylyi Crc
-
-
-
-
?
3,4-dihydroxybenzoate + O2
3-carboxy-cis,cis-muconate
show the reaction diagram
Hydrogenophaga palleronii S1
-
-
-
?
3,4-dihydroxybenzoate + O2
3-carboxy-cis,cis-muconate
show the reaction diagram
Acinetobacter calcoaceticus 80-1
-
-
-
?
3,4-dihydroxybenzoate + O2
3-carboxy-cis,cis-muconate
show the reaction diagram
Pseudomonas aeruginosa B-10
-
very low activity with: 3,4-dihydroxyphenylacetic acid, very low activity with 3,4-dihydroxymandelic acid, very low activity with 4'-methylcatechol
-
?
3,4-dihydroxybenzoate + O2
3-carboxy-cis,cis-muconate
show the reaction diagram
Pseudomonas fluorescens A.3.12.
-
no other substrate found
-
ir
3,4-dihydroxybenzoate + O2
3-carboxy-cis,cis-muconate
show the reaction diagram
-
-
-
-
?
3,4-dihydroxybenzoate + O2
3-carboxy-cis,cis-muconate
show the reaction diagram
Stenotrophomonas maltophilia KB2
-
-
-
-
?
3,4-dihydroxybenzoate + O2
3-carboxy-cis,cis-muconate
show the reaction diagram
Acinetobacter baylyi ADPU54
-
-
-
-
?
3,4-dihydroxybenzoate + O2
?
show the reaction diagram
-
spectroscopic and electronic structure study of the enzyme-substrate complex
-
-
?
3,4-dihydroxybenzoate + O2
beta-carboxy-cis,cis-muconate
show the reaction diagram
-
-
-
-
?
3,4-dihydroxymandelic acid + O2
?
show the reaction diagram
Nocardia erythropolis
-
at 5.4% the rate of protocatechuic acid oxidation
-
-
?
3,4-dihydroxymandelic acid + O2
?
show the reaction diagram
Hydrogenophaga palleronii, Hydrogenophaga palleronii S1
-
protocatechuate 3,4-dioxygenase II
-
-
?
3,4-dihydroxyphenylacetate + O2
?
show the reaction diagram
-
-
-
-
?
3,4-dihydroxyphenylacetate + O2
?
show the reaction diagram
-
-
-
-
-
3,4-dihydroxyphenylacetate + O2
?
show the reaction diagram
-
-
-
-
?
3,4-dihydroxyphenylacetate + O2
?
show the reaction diagram
-
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
-
-
-
3,4-dihydroxyphenylalanine + O2
?
show the reaction diagram
Nocardia erythropolis
-
at 10.7% the rate of protocatechuic acid oxidation
-
-
?
3,4-dihydroxyphenylpropionic acid + O2
?
show the reaction diagram
-
-
-
-
-
3-(3,4-dihydroxyphenyl)propionate + O2
?
show the reaction diagram
-
protocatechuate 3,4-dioxygenase II
-
-
?
3-methylcatechol + O2
2-methylmuconate
show the reaction diagram
-
-
-
?
3-methylcatechol + O2
2-methylmuconate
show the reaction diagram
-
at 0.4% the rate of protocatechuic acid oxidation
-
?
3-methylcatechol + O2
2-methylmuconate
show the reaction diagram
Nocardia erythropolis
-
at 14.4% the rate of protocatechuic acid oxidation
-
?
3-methylcatechol + O2
2-methylmuconate
show the reaction diagram
-
at 5% the rate of protocatechuic acid oxidation
-
?
3-methylcatechol + O2
2-methylmuconate
show the reaction diagram
Agrobacterium tumefaciens S2, Hydrogenophaga palleronii S1
-
-
-
?
4-cresol + O2
?
show the reaction diagram
-
-
-
-
?
4-methylcatechol + O2
3-methyl-cis,cis-muconate
show the reaction diagram
Nocardia erythropolis
-
at 3.1% the rate of protocatechuic acid oxidation
-
?
4-methylcatechol + O2
3-methyl-cis,cis-muconate
show the reaction diagram
-
at very low rates
-
?
4-methylcatechol + O2
?
show the reaction diagram
-
-
-
-
?
4-sulfocatechol + O2
3-sulfomuconate
show the reaction diagram
-
catalyzed by protocatechuate 3,4-dioxgenase type II only
-
?
5-fluoro-protocatechuic acid + O2
5-fluoro-3-carboxy-cis,cis-muconate
show the reaction diagram
-
at 2.1% the rate of protocatechuic acid oxidation
-
?
6-chloro-protocatechuate + O2
6-chloro-3-carboxy-cis,cis-muconate
show the reaction diagram
-
at 4.3% the rate of protocatechuic acid oxidation
-
?
catechin + O2
?
show the reaction diagram
-
-
-
-
?
catechol + O2
cis,cis-muconate
show the reaction diagram
-
reaction of EC 1.13.11.1
-
-
?
catechol + O2
muconate
show the reaction diagram
Geobacillus sp.
-
-
-
-
?
catechol + O2
muconate
show the reaction diagram
-
at 0.4% the rate of protocatechuic acid oxidation
-
?
catechol + O2
muconate
show the reaction diagram
-
at 3% the rate of protocatechuic acid oxidation
-
?
catechol + O2
muconate
show the reaction diagram
Nocardia erythropolis
-
at 33.8% the rate of protocatechuic acid oxidation
-
?
catechol + O2
muconate
show the reaction diagram
Agrobacterium tumefaciens S2, Hydrogenophaga palleronii S1
-
-
-
?
catechol + O2
muconate
show the reaction diagram
Pseudomonas aeruginosa B-10
-
at 0.4% the rate of protocatechuic acid oxidation
-
?
gallate + O2
?
show the reaction diagram
-
-
-
-
?
protocatechuate + O2
beta-carboxy-cis-cis-muconate
show the reaction diagram
-
-
-
-
?
protocatechuate + O2
3-carboxy-cis,cis-muconate
show the reaction diagram
Stenotrophomonas maltophilia, Stenotrophomonas maltophilia KB2
-
-
-
-
?
protocatechuic acid + O2
3-carboxy-cis,cis-muconic acid
show the reaction diagram
Geobacillus sp.
-
-
-
-
?
pyrogallol + O2
?
show the reaction diagram
-
-
-
-
?
pyrogallol + O2
?
show the reaction diagram
-
at 10% the rate of protocatechuic acid oxidation
-
-
?
pyrogallol + O2
?
show the reaction diagram
Nocardia erythropolis
-
at 36% the rate of protocatechuic acid oxidation
-
-
?
pyrogallol + O2
?
show the reaction diagram
Pseudomonas aeruginosa, Pseudomonas aeruginosa B-10
-
at 0.4% the rate of protocatechuic acid oxidation
-
-
?
pyrogallol + O2
?
show the reaction diagram
-
-
-
-
?
trans-3,4-dihydroxycinnamate + O2
?
show the reaction diagram
-
-
-
-
?
trans-3,4-dihydroxycinnamate + O2
?
show the reaction diagram
-
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
-
-
-
L-DOPA + O2
?
show the reaction diagram
-
-
-
-
?
additional information
?
-
A8I4B3, A8I4B7, -
benzoate induces expression of protocatechuate 3,4-dioxygenase
-
-
-
additional information
?
-
Geobacillus sp.
-
no activity with gentisate
-
-
-
additional information
?
-
Geobacillus sp.
-
enzyme involved in last ring fission in naphtalene degradation, intermediates confirm proceeding through protocatechuic acid via ortho-claevage pathway
-
-
-
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
?
-
-
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
?
-
A8I4B3, A8I4B7
benzoate induces expression of protocatechuate 3,4-dioxygenase
-
-
-
additional information
?
-
Stenotrophomonas maltophilia KB2
-
strain KB2 degrades 13 mM 3,4-dihydroxybenzoate, 10 mM benzoic acid and 12 mM phenol within 24 h of incubation
-
-
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
3,4-dihydroxybenzoate + O2
3-carboxy-cis,cis-muconate
show the reaction diagram
-
-
-
-
3,4-dihydroxybenzoate + O2
3-carboxy-cis,cis-muconate
show the reaction diagram
-
-
-
-
?
3,4-dihydroxybenzoate + O2
3-carboxy-cis,cis-muconate
show the reaction diagram
-
-
-
-
?
3,4-dihydroxybenzoate + O2
3-carboxy-cis,cis-muconate
show the reaction diagram
-
-
-
-
?
3,4-dihydroxybenzoate + O2
3-carboxy-cis,cis-muconate
show the reaction diagram
-
-
-
?
3,4-dihydroxybenzoate + O2
3-carboxy-cis,cis-muconate
show the reaction diagram
Leifsonia sp.
-
-
-
-
?
3,4-dihydroxybenzoate + O2
3-carboxy-cis,cis-muconate
show the reaction diagram
-
-
-
-
?
protocatechuate + O2
beta-carboxy-cis-cis-muconate
show the reaction diagram
-
-
-
-
?
protocatechuic acid + O2
3-carboxy-cis,cis-muconic acid
show the reaction diagram
Geobacillus sp.
-
-
-
-
?
3,4-dihydroxybenzoate + O2
3-carboxy-cis,cis-muconate
show the reaction diagram
Stenotrophomonas maltophilia KB2
-
-
-
-
?
additional information
?
-
A8I4B3, A8I4B7, -
benzoate induces expression of protocatechuate 3,4-dioxygenase
-
-
-
additional information
?
-
Geobacillus sp.
-
enzyme involved in last ring fission in naphtalene degradation, intermediates confirm proceeding through protocatechuic acid via ortho-claevage pathway
-
-
-
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
?
-
-
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
?
-
A8I4B3, A8I4B7
benzoate induces expression of protocatechuate 3,4-dioxygenase
-
-
-
additional information
?
-
Stenotrophomonas maltophilia KB2
-
strain KB2 degrades 13 mM 3,4-dihydroxybenzoate, 10 mM benzoic acid and 12 mM phenol within 24 h of incubation
-
-
-
METALS and IONS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
Cu2+
Leifsonia sp.
-
increases enzyme activity for both cell-free and immobilized extracts by 8% and 44%, respectively
Fe
-
enzyme contains 7.3 g-atom of nonheme iron per mol of enzyme
Fe
-
5 irons per enzyme molecule
Fe
-
external addition of FeSO4 is absolutely essential, other metal ions cannot replace Fe2+
Fe
-
6.6 iron atoms per holoenzyme of 700000 Da
Fe
-
1.3-1.5 g-atom Fe3+ per mol of enzyme
Fe
-
7.27 g-atoms iron per mol enzyme
Fe
-
10 g-atoms per mol enzyme
Fe
-
5 Fe atoms per holoenzyme
Fe
-
enzyme contains 4 Fe3+
Fe2+
-
activates by 29% at 3 mM
Fe3+
-
presence of a alphabetaFe3+ protomer in a variety of stoichiometries
Fe3+
Leifsonia sp.
-
increases enzyme activity for both cell-free and immobilized extracts by 16% and 99%, respectively
INHIBITORS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
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
2,3-Dihydroxybenzoate
-
-
2,3-Dihydroxybenzoate
-
-
2,4-dihydroxybenzoic acid
-
competitive inhibition
2,5-Dihydroxybenzoate
-
-
2-Fluoro-4-hydroxybenzoate
-
-
2-Hydroxyisonicotinic acid N-oxide
-
0.000025 mM, 50% inactivation after 10 min, most potent inhibitor
2-Hydroxyisonicotinic acid N-oxide
-
-
2-Hydroxypyridine N-oxide
-
-
3,4-Dihydroxyacetophenone
-
-
3,4-Dihydroxyacetophenone
-
-
3,4-dihydroxybenzoate
-
-
3,4-dihydroxycinnamic acid
-
-
3,4-Dihydroxyphenylacetate
-
-
3,4-dihydroxyphenylacetic acid
-
-
3,4-dihydroxyphenylacetic acid
-
0.006 mM, 92% inhibition
3,4-dihydroxyphenylacetic acid
-
-
3,4-Dihydroxyphenylpropionate
-
-
3,5-Dichloro-4-hydroxybenzoate
-
-
3-Bromo-4-hydroxybenzoate
-
-
3-Chloro-4-hydroxybenzoate
-
-
3-Fluoro-4-hydroxybenzoate
-
-
3-Fluoro-4-hydroxybenzoate
-
-
3-Hydroxybenzoate
-
-
3-Hydroxybenzoate
-
optical titration at 25C
3-hydroxyphenylacetic acid
-
-
3-Nitrophenol
-
0.006 mM, 56% inhibition
4-(dimethylamino)benzaldehyde
-
-
4-Fluoro-3-hydroxybenzoate
-
-
4-Fluoro-3-hydroxybenzoate
-
-
4-hydroxybenzoate
-
-
4-hydroxybenzoate
-
-
4-hydroxybenzoate
-
-
4-hydroxybenzoate
-
optical titration at 25C
4-hydroxymercuribenzoate
-
-
4-hydroxymercuribenzoate
-
0.1 mM, complete inhibition
4-hydroxymercuribenzoate
-
0.006 mM, 80% inhibition
4-hydroxyphenylacetic acid
-
-
4-Methylcatechol
-
0.006 mM, 59% inhibition
4-Methylcatechol
-
-
4-nitrocatechol
-
0.006 mM, complete inhibition
4-nitrocatechol
-
-
4-sulfocatechol
-
competitive vs. protocatechuate
Al3+
-
EDTA salt, 0.01 mM, complete inhibition
Al3+
-
inhibits 66.5% at 3 mM
alpha-chloro-3,4-dihydroxyacetophenone
-
-
Ca2+
Nocardia erythropolis
-
-
catechol
-
0-006 mM, 63% inhibition
Cd2+
-
inhibits 40% at 3 mM
Co2+
-
inhibits 48.5% at 3 mM
Cu2+
-
inhibits 70% at 3 mM
Fe3+
-
inhibits 77% at 3 mM
-
Ferrous ammonium sulfate
-
-
Hg2+
Nocardia erythropolis
-
0.1 mM, 45% inhibition
iodoacetate
-
5 mM, 20% inhibition
iodoacetate
-
0.1 mM, complete inhibition
iodoacetate
Nocardia erythropolis
-
-
isonicotinic acid N-oxide
-
-
KF
-
50 mM, 34% inhibition
N-ethylmaleimide
-
0.1 mM, complete inhibition
N-ethylmaleimide
Nocardia erythropolis
-
-
Na2HAsO4
Nocardia erythropolis
-
0.1 mM, 50% inhibition
Ni2+
-
EDTA salt, 0.01 mM, 72% inhibition
Ni2+
-
inhibits 17% at 3 mM
Nickel ammonium sulfate
-
-
o-Chloranil
-
0.006 mM, 51% inhibition
p-chloromercuribenzoate
-
0.05 mM, 47% inhibition
Pb2+
Nocardia erythropolis
-
-
Protocatechualdehyde
-
competitive inhibitor
Protocatechualdehyde
-
-
Protocatechualdehyde
-
0.006 mM, 54% inhibition
Protocatechualdehyde
-
-
protocatechuate
-
competitive vs. 4-sulfocatechol
Protocatechuic acid methyl ester
-
-
Zn2+
-
inhibits 49% at 3 mM
Mn2+
-
inhibits 19% at 3 mM
additional information
-
not inhibited by arsenate, iodoacetate, semicarbazide, N-phenylmaleimide, fluoride or cyanide
-
additional information
-
not inhibited by iodoacetamide, 5,5'-dithiobis(2-nitrobenzoate), mercaptoethanol, dithiothreitol, glutathione, o-phenanthroline and H2O2
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
Na2SO4
-
nonessential activator
KM VALUE [mM]
KM VALUE [mM] Maximum
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.0029
-
3,4-dihydroxybenzoate
-
mutant enzyme Y447H
0.007
-
3,4-dihydroxybenzoate
Geobacillus sp.
-
-
0.017
-
3,4-dihydroxybenzoate
-
pH 7.5, 38C
0.03
-
3,4-dihydroxybenzoate
-
wild-type enzyme
0.036
-
3,4-dihydroxybenzoate
-
mutant enzyme R457S
0.038
-
3,4-dihydroxybenzoate
-
mutant enzyme DELTA319-322
0.078
-
3,4-dihydroxybenzoate
-
wild-type enzyme
0.142
-
3,4-dihydroxybenzoate
-
mutant enzyme R133H
0.014
-
4-Cresol
-
pH 7.5, 38C
0.014
-
4-Methylcatechol
-
pH 7.5, 38C
0.042
-
4-sulfocatechol
-
protocatechuate 3,4-dioxgenase type II
0.058
-
4-sulfocatechol
-
protocatechuate 3,4-dioxgenase type II
0.014
-
caffeic acid
-
pH 7.5, 38C
0.014
-
catechin
-
pH 7.5, 38C
0.014
-
catechol
-
pH 7.5, 38C
0.033
-
catechol
Geobacillus sp.
-
-
0.04
-
FeSO4
-
-
0.002
-
gallate
-
pH 7.5, 38C
0.006
-
O2
-
Y447H mutant enzyme
0.04
-
O2
-
native enzyme
0.056
-
O2
-
recombinant enzyme
0.056
-
O2
-
wild-type enzyme
0.07
-
O2
-
at pH 8.3, spectrophotometric
0.09
-
O2
-
at pH 8.3, oxygen electrode
0.002
-
protocatechuate
-
at pH 8.3, spectrophotometric
0.0029
-
protocatechuate
-
Y447H mutant enzyme
0.003
-
protocatechuate
-
at pH 8.3, oxygen electrode
0.0175
-
protocatechuate
-
-
0.018
-
protocatechuate
-
-
0.029
-
protocatechuate
-
recombinant enzyme
0.03
-
protocatechuate
-
-
0.03
-
protocatechuate
-
native enzyme
0.054
-
protocatechuate
-
protocatechuate 3,4-dioxgenase type I
0.058
-
protocatechuate
-
protocatechuate 3,4-dioxgenase type II
0.0714
-
protocatechuate
-
-
0.1
-
protocatechuate
Nocardia erythropolis
-
-
0.125
-
protocatechuate
-
-
0.205
-
protocatechuate
-
protocatechuate 3,4-dioxgenase type II
0.33
-
protocatechuate
-
-
0.014
-
pyrogallol
-
pH 7.5, 38C
0.014
-
L-Dopa
-
pH 7.5, 38C
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
-
TURNOVER NUMBER [1/s]
TURNOVER NUMBER MAXIMUM[1/s]
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.007
-
3,4-dihydroxybenzoate
-
mutant enzyme Y408H
0.01
-
3,4-dihydroxybenzoate
-
mutant enzyme Y408E
0.09
-
3,4-dihydroxybenzoate
-
mutant enzyme R457S
0.12
-
3,4-dihydroxybenzoate
-
mutant enzyme Y447H
0.13
-
3,4-dihydroxybenzoate
-
mutant enzyme Y408C
0.24
-
3,4-dihydroxybenzoate
-
mutant enzyme R133H
29
-
3,4-dihydroxybenzoate
-
mutant enzyme DELTA319-322
68
-
3,4-dihydroxybenzoate
-
wild-type enzyme
120
-
3,4-dihydroxybenzoate
-
wild-type enzyme
417
-
3,4-dihydroxybenzoate
-
-
758
-
3,4-dihydroxybenzoate
-
-
14.6
-
4-sulfocatechol
-
protocatechuate 3,4-dioxgenase type II
0.007
-
O2
-
mutant enzyme Y408H
0.01
-
O2
-
mutant enzyme Y408E
0.09
-
O2
-
mutant enzyme R457S
0.12
-
O2
-
mutant enzyme Y447H
0.13
-
O2
-
mutant enzyme Y408C
0.24
-
O2
-
mutant enzyme R133H
29
-
O2
-
mutant enzyme DELTA319-322
68
-
O2
-
wild-type enzyme
120
-
O2
-
wild-type enzyme
0.007
-
protocatechuate
-
for the mutant enzyme Y408H, at 25C 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 25C 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 25C in 50 mM Tris and 2 mM beta-mercaptoethanol, pH 8.5 under saturating protocatechuate and O2 conditions
0.12
-
protocatechuate
-
Y447H mutant enzyme
0.13
-
protocatechuate
-
for the mutant enzyme Y408C, at 25C in 50 mM Tris and 2 mM beta-mercaptoethanol, pH 8.5 under saturating protocatechuate and O2 conditions
18.8
-
protocatechuate
-
protocatechuate 3,4-dioxgenase type II
64.8
-
protocatechuate
-
oxygen electrode at pH 8.3
68
-
protocatechuate
-
recombinant enzyme
70
-
protocatechuate
-
native enzyme
74.7
-
protocatechuate
-
spectrophotometric at pH 8.3
89.5
-
protocatechuate
-
protocatechuate 3,4-dioxgenase type I
100
-
protocatechuate
-
for the wild-type enzyme, at 25C in 50 mM Tris and 2 mM beta-mercaptoethanol, pH 8.5 under saturating protocatechuate and O2 conditions
Ki VALUE [mM]
Ki VALUE [mM] Maximum
INHIBITOR
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.29
-
2,5-Dihydroxybenzoate
-
-
0.01
-
2-Fluoro-4-hydroxybenzoate
-
-
0.0068
-
2-Hydroxyisonicotinic acid N-oxide
-
apparent Ki, irreversible binding
3.9
-
2-Hydroxypyridine N-oxide
-
apparent Ki, inhibition is not freely reversible
0.13
-
3,4-Dihydroxyacetophenone
-
-
0.013
-
3,4-Dihydroxyphenylacetate
-
-
0.041
-
3,4-dihydroxyphenylacetic acid
-
-
0.018
-
3,4-Dihydroxyphenylpropionate
-
-
0.044
-
3,5-Dichloro-4-hydroxybenzoate
-
-
0.018
-
3-Bromo-4-hydroxybenzoate
-
-
0.0032
-
3-Chloro-4-hydroxybenzoate
-
-
0.0003
-
3-Fluoro-4-hydroxybenzoate
-
at pH 7.0
0.0005
-
3-Fluoro-4-hydroxybenzoate
-
-
0.01
-
3-Fluoro-4-hydroxybenzoate
-
-
0.004
-
3-Hydroxybenzoate
-
-
0.3
-
4-Fluoro-3-hydroxybenzoate
-
-
0.8
-
4-Fluoro-3-hydroxybenzoate
-
-
0.087
-
4-hydroxybenzoate
-
at pH 8.4
0.1
-
4-hydroxybenzoate
-
-
0.24
-
4-hydroxybenzoate
-
-
0.44
-
4-Methylcatechol
-
-
0.0006
-
4-nitrocatechol
-
-
0.0048
-
4-nitrocatechol
-
-
0.032
-
4-sulfocatechol
-
protocatechuate 3,4-dioxgenase type II
0.053
-
4-sulfocatechol
-
protocatechuate 3,4-dioxgenase type II
0.17
-
alpha-chloro-3,4-dihydroxyacetophenone
-
-
0.7
-
catechol
-
-
1.3
-
isonicotinic acid N-oxide
-
-
0.001
-
Protocatechualdehyde
-
-
0.014
-
Protocatechualdehyde
-
-
0.014
-
Protocatechualdehyde
-
-
0.073
-
Protocatechualdehyde
-
-
0.066
-
protocatechuate
-
protocatechuate 3,4-dioxgenase type II
0.102
-
protocatechuate
-
protocatechuate 3,4-dioxgenase type II
0.4
-
protocatechuic acid methylester
-
-
0.003
-
vanillate
-
-
0.02
-
Isovanillate
-
-
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
-
SPECIFIC ACTIVITY [µmol/min/mg]
SPECIFIC ACTIVITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
0.0081
-
-
-
0.113
-
-
activity in E. coli cell extracts expressing the recombinant enzyme
0.31
-
Roseobacter sp.
-
Rosebacter isolate GAI-16, enzyme activity in cell extracts
0.34
-
-
soluble enzyme
0.39
-
-
enzyme immobilized on glass-beads
0.46
-
-
enzyme activity in cell extracts
0.73
-
Roseobacter sp.
-
Rosebacter isolate S25com04, enzyme activity in cell extracts
0.88
-
Roseobacter sp.
-
Rosebacter isolate IC4, enzyme activity in cell extracts
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
16.7
-
-
type II protocatechuate 3,4-dioxygenase, oxidation of 4-sulfocatechol
34.2
-
Geobacillus sp.
-
-
37.2
-
-
recombinant enzyme
105
-
-
type I protocatechuate 3,4-dioxygenase
121.7
-
-
purified native enzyme, pH 7.5, 38C
350
-
Geobacillus sp.
-
activity at the 10th h of the exponential growth phase
additional information
-
Nocardia erythropolis
-
379 mg O2/min/ml
pH OPTIMUM
pH MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
5.2
-
-
completely inactive above pH 6.5
7.3
8.5
-
immobilized enzyme
7.5
-
-
immobilized enzyme
7.5
-
Leifsonia sp.
-
assay at
8
-
-
free enzyme
8
-
Nocardia erythropolis
-
-
8
-
Geobacillus sp.
-
-
8.3
8.4
-
free enzyme
8.5
-
-
maximum activity for the wild type enzyme and the mutants enymes Y408C, Y408F, Y408E and Y408H
pH RANGE
pH RANGE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
4
9
Leifsonia sp.
-
activity range, profile overview
6
9
Geobacillus sp.
-
pH 6.0: about 40% of maximal activity, pH 9.0: about 65% of maximal activity
6.5
8
Leifsonia sp.
-
80% of maximal activity within this range
additional information
-
-
activity increases steadily up to pH 9.0, above pH 9.0 the enzyme begins to undergo spontaneous oxidation
TEMPERATURE OPTIMUM
TEMPERATURE OPTIMUM MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
35
-
Leifsonia sp.
-
optimum for the cell-free enzyme extract
37
-
-
free enzyme, 9% activity at 60C, no activity at 80C
40
-
-
-
40
-
Nocardia erythropolis
-
-
45
-
-
immobilized enzyme
55
-
Leifsonia sp.
-
optimum for the immobilized enzyme extract
60
-
Geobacillus sp.
-
-
TEMPERATURE RANGE
TEMPERATURE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
45
80
-
immobilized enzyme: 78% activity at 60C, 75% activity at 80C
50
70
Geobacillus sp.
-
50C: about 55% of maximal activity, 70C: about 60% of maximal activity
SOURCE TISSUE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
SOURCE
-
monocyclic hydrocarbons, 4-hydroxybenzoic acid, protocatechuic acid and vanillic acid
Manually annotated by BRENDA team
Stenotrophomonas maltophilia KB2
-
monocyclic hydrocarbons, 4-hydroxybenzoic acid, protocatechuic acid and vanillic acid
-
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
-
localized in 100000 g supernatant, completely absent in mitochondrial, microsomal and chloroplast fraction
Manually annotated by BRENDA team
PDB
SCOP
CATH
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)
MOLECULAR WEIGHT
MOLECULAR WEIGHT MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
150000
-
-
gel filtration
150000
-
Nocardia erythropolis
-
gel filtration
189900
-
-
gel filtration
190000
-
-
gel filtration
202000
-
-
meniscus depletion
204000
-
-
sedimentation equilibrium
220000
-
-
gel filtration
315000
-
-
analytical ultracentrifugation
480000
-
-
ultracentrifugation
480000
-
Geobacillus sp.
-
gel filtration
510000
-
-
gel filtration
590000
-
-
ultracentrifugation
677000
-
-
sedimentation velocity analysis
700000
-
-
diffusion data
783100
-
-
calculated
SUBUNITS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
?
-
x * 21900 + x * 26700, deduced from nucleotide sequences of putative alpha and beta subunits
?
-
x * 27097 + x * 21502, deduced from nucleotide sequence
?
-
x * 26000 + x * 22000, 2D-PAGE
?
-
x * 26000 + x * 22000, 2D-PAGE
-
decamer
-
alpha,beta, 5 * 23300 + 5 * 25500, SDS-PAGE
decamer
-
alpha,beta, 5 * 22500 + 5 * 40000, SDS-PAGE
hexamer
-
trimer of dimers, (alphabeta)3, 3 * 29000, alpha subunit, + 3 * 34300, beta-subunit, SDS-PAGE
octamer
-
alpha,beta, 4 * 26500 + 4 * 23000
octamer
-
alpha,beta, 4 * 29000 + 4 * 26500, SDS-PAGE
octamer
-
alpha,beta, 4 * 23000 + 4 * 26500, SDS-PAGE
octamer
-
8 * 97832, calculated
tetramer
-
alpha,beta, 2 * 22280 + 2 * 26630, holoenzyme probably contains 6 alpha2,beta2 tetramers, amino acid sequence
hexamer
-
trimer of dimers, (alphabeta)3, 3 * 29000, alpha subunit, + 3 * 34300, beta-subunit, SDS-PAGE
-
additional information
-
amino acid sequence of alpha-subunit, 203 amino acids
additional information
-
amino acid sequence of beta-subunit, 238 amino acids
additional information
-
quarternary structure
additional information
-
crystal structure, dodecameric enzyme
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
no glycoprotein
-
carbohydrate contributes less than 0.2% to the mass of the holoenzyme
Crystallization/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
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 18C, 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
-
X-ray crystallography
-
free interface diffusion cell, equal volumes of concentrated enzyme and ammonium sulfate solutions ranging from 37-47% saturation, 4C, 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 4C, recombinant wild-type and Y447H mutant enzyme
-
pH STABILITY
pH STABILITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
6
9.6
-
24C, 1 week, less than 10% loss of activity
7.6
8.6
Nocardia erythropolis
-
most stable
8.5
-
-
60C, 1 h, 50% loss of activity
8.5
-
-
50C, 90 min stable
TEMPERATURE STABILITY
TEMPERATURE STABILITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
5
-
-
stable for long periods
20
50
-
purified enzyme, 50 min, pH 7.5, relatively stable
42
-
Nocardia erythropolis
-
1 h, stable below
50
-
-
3 min, inactivation
50
-
-
pH 8.5, 90 min, stable
50
-
Nocardia erythropolis
-
1 h, unstable above
60
-
-
5 min, about 5% loss of activity
60
-
-
immobilized enzyme shows slightly enhanced stability
60
-
-
5 min, stable
60
-
-
pH 8.5, 1 h, 50% loss of activity
60
-
-
pH 8.5, 10 min, 90% loss of activity
60
-
Geobacillus sp.
-
half-life is 40 min
65
-
-
soluble and immobilized enzyme, 15 min, complete loss of activity
70
-
-
purified enzyme, 20 min, pH 7.5, loss of 95% activity
GENERAL STABILITY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
no loss of activity over a period of 78 d under aerobic conditions at 15C at pH 7.0, pH 8.0 or pH 9.0
-
lyophilization, stable
-
UV irradiation: 38000 ergs per m2, inactivates
-
enzyme immobilized on porous glass beads is stable over wide ranges of pH and temperature
-
immobilization: slight increase of thermal stability, soluble and immobilized enzyme exhibit substantial activity in 2 M urea
-
inactivated by reducing agents
-
STORAGE STABILITY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
4C, pH 8.5, 6 months
-
-20C, stable for several years
-
5C, long periods, no loss in activity
-
0-3C, several weeks, no loss in activity
-
stable only 7 days
-
Purification/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
ammonium sulfate, DEAE-cellulose, Sephadex G-200
-
ammonium sulfate, Q-Sepharose, partial purification
-
recombinant enzyme
-
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
-
ammonium sulfate, DEAE-cellulose, Bio-gel agarose, DEAE-cellulose
-
ammonium sulfate, DEAE-Sepharose, Phenyl-Sepharose
-
-
Geobacillus sp.
-
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 -20C
Geobacillus sp.
-
Q-Sepharose FF, ammonium sulfate, Superdex 200, Fractogel, type II protocatechuate 3,4-dioxygenase
-
-
Nocardia erythropolis
-
native enzyme 296.8fold by ammonium sulfate fractionation, anion exchange and hydrophobic interaction chromatography
-
ammonium sulfate, DEAE-Sepharose, octyl-Sepharose, Phenyl-sepharose, recombinant enzyme
-
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
-
ammonium sulfate, Sephacryl S 300, DEAE-Sephadex A50
-
ammonium sulfate, DEAE-cellulose
-
Cloned/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
expression of wild-type, W153V, R142K and R142/W153V double mutant of protocatechuate 3,4-dioxygenase I in Escherichia coli
-
expressed in Escherichia coli, the genetic locus ncg12314-ncg12315 of the Corynebacterium glutamicum encodes a putative protocatechuate 3,4-dioxygenase
-
expression of protocatechuate 3,4-dioxygenase II in Escherichia coli
-
phylogenetic tree
Leifsonia sp.
-
expression of wild-type and Y447H mutant enzyme in Pseudomonas fluorescens
-
the enzyme containing the Y408E mutation is expressed in Pseudomonas fluorescens; the enzymes containing the Y408C, Y408F and Y408H mutation are expressed in Escherichia coli
-
expressed in Escherichia coli
-
expression in Escherichia coli
-
EXPRESSION
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
Crc induces carbon catabolite repression of protocatechuate 3,4-dioxygenase. Crc is not involved in carbon catabolite repression of the protocatechuate 3,4-dioxygenase operon (pca-qui) at the transcriptional level. Addition of acetate and succinate to medium containing a carbon source leads to repression of protocatechuate 3,4-dioxygenase by 95%, whereas other organic acids, like pyruvate, do not have a repressing effect
-
Crc induces carbon catabolite repression of protocatechuate 3,4-dioxygenase. Crc is not involved in carbon catabolite repression of the protocatechuate 3,4-dioxygenase operon (pca-qui) at the transcriptional level. Addition of acetate and succinate to medium containing a carbon source leads to repression of protocatechuate 3,4-dioxygenase by 95%, whereas other organic acids, like pyruvate, do not have a repressing effect
Acinetobacter baylyi ADPU54, Acinetobacter baylyi Crc
-
-
protocatechuate induces PcaHG activity and PcaO regulates the key step in aromatic cleavage during PCA degradation at the pcaHG transcriptional level
-
ENGINEERING
ORGANISM
UNIPROT ACCESSION NO.
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
-
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
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
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
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.
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
-
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
-
greatly reduced rate of protocatechuate oxygenation
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
Acinetobacter baylyi ADP1
-
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.
-
R153V
-
protocatechuate 3,4-dioxygenase I gain of function mutation, mutant enzyme oxidizes 4-sulfocatechol
additional information
Leifsonia sp.
-
immobilization of the enzyme, the immobilized extract exhibited higher enzyme activity than the cell-free extract in the presence of trace elements and cations
APPLICATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
analysis
-
use in flushing and scrubbing oxygen out of instruments such as stopped-flow spectrophotometers
environmental protection
Geobacillus sp.
-
the purified enzyme can be used in bioremediation of polluted groundwater or soil contaminated with various aromatic compounds ranging from monocyclic to polycyclic
degradation
-
because of broad spectrum of dioxygenases types that Stenotrophomonas maltophilia KB2 can exhibit, this strain appears to be very powerful and useful tool in the biotreatment of wastewaters and in soil decontamination
degradation
Stenotrophomonas maltophilia KB2
-
because of broad spectrum of dioxygenases types that Stenotrophomonas maltophilia KB2 can exhibit, this strain appears to be very powerful and useful tool in the biotreatment of wastewaters and in soil decontamination
-