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 hide
1.13.11.3
-
RECOMMENDED NAME
GeneOntology No.
protocatechuate 3,4-dioxygenase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
3,4-dihydroxybenzoate + O2 = 3-carboxy-cis,cis-muconate
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
oxidation
-
-
-
-
redox reaction
-
-
-
-
reduction
-
-
-
-
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
protocatechuate degradation II (ortho-cleavage pathway)
-
-
4-hydroxymandelate degradation
-
-
gallate degradation
-
-
Benzoate degradation
-
-
Polycyclic aromatic hydrocarbon degradation
-
-
Metabolic pathways
-
-
Microbial metabolism in diverse environments
-
-
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.
CAS REGISTRY NUMBER
COMMENTARY hide
9029-47-4
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
derivative of ATCC 14987
-
-
Manually annotated by BRENDA team
derivative of ATCC 14987
-
-
Manually annotated by BRENDA team
enzyme is induced by growth on p-hydroxybenzoate
-
-
Manually annotated by BRENDA team
enzyme is induced by growth on p-hydroxybenzoate
-
-
Manually annotated by BRENDA team
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 II
-
-
Manually annotated by BRENDA team
-
-
-
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
nduced in cells grown on 4-hydroxybenzoate
-
-
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
subunit alpha and subunit beta
A0A193DXA9 and A0A193DXP2
UniProt
Manually annotated by BRENDA team
beta- and alpha-subunit
Q0SH27 AND Q0SH26
UniProt
Manually annotated by BRENDA team
isolates GAI-16, S25com04 and IC4, enzyme activity is induced by growth on p-hydroxybenzoate
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
enzyme activity is induced by growth on p-hydroxybenzoate
-
-
Manually annotated by BRENDA team
and strains Y3F, Y4I
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
metabolism
additional information
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
2,3-dihydroxybenzoate + O2
?
show the reaction diagram
2,4-dihydroxybenzoate + O2
?
show the reaction diagram
2,5-dihydroxybenzoate + O2
?
show the reaction diagram
2,6-dihydroxybenzoate + O2
?
show the reaction diagram
3,4-dihydroxybenzoate + O2
3-carboxy-cis,cis-muconate
show the reaction diagram
3,4-dihydroxybenzoate + O2
?
show the reaction diagram
3,4-dihydroxybenzoate + O2
beta-carboxy-cis,cis-muconate
show the reaction diagram
-
-
-
-
?
3,4-dihydroxyhydrocinnamic acid + O2
?
show the reaction diagram
3,4-dihydroxymandelic acid + O2
?
show the reaction diagram
3,4-dihydroxyphenylacetate + O2
?
show the reaction diagram
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
-
-
-
-
-
3,5-dihydroxybenzoate + 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
4-cresol + O2
?
show the reaction diagram
-
-
-
-
?
4-hydroxybenzoic acid + O2
?
show the reaction diagram
4-methylcatechol + O2
3-methyl-cis,cis-muconate
show the reaction diagram
4-methylcatechol + O2
?
show the reaction diagram
-
-
-
-
?
4-sulfocatechol + O2
3-sulfomuconate
show the reaction diagram
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
-
?
caffeic acid + O2
?
show the reaction diagram
catechin + O2
?
show the reaction diagram
-
-
-
-
?
catechol + O2
cis,cis-muconate
show the reaction diagram
catechol + O2
muconate
show the reaction diagram
gallate + O2
?
show the reaction diagram
-
-
-
-
?
L-DOPA + O2
?
show the reaction diagram
-
-
-
-
?
protocatechuate + 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
-
-
-
-
?
pyrogallol + O2
?
show the reaction diagram
trans-3,4-dihydroxycinnamate + O2
?
show the reaction diagram
vanillic acid + O2
?
show the reaction diagram
additional information
?
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
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
-
-
-
-
?
additional information
?
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
2,2'-dipyridyl
I0DHJ0 AND I0DHJ1
activates the enzyme immobilized on calcium alginate, but inhibits the free enzyme and the enzyme immobilized on glyoxyl agarose, at 0.3 mM
Al3+
I0DHJ0 AND I0DHJ1
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
I0DHJ0 AND I0DHJ1
activates the enzyme immobilized on calcium alginate, but inhibits the free enzyme and the enzyme immobilized on glyoxyl agarose, at 0.3 mM
Fe2+
-
activates by 29% at 3 mM
additional information
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
(2E)-3-(3,4-dimethoxyphenyl)-N-hydroxyprop-2-enamide
Q0SH27 AND Q0SH26
over 90% inhibition at 0.2 mM
(2E)-N-hydroxy-3-(4-methoxyphenyl)prop-2-enamide
Q0SH27 AND Q0SH26
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
2,2'-dipyridyl
2,3-Dihydroxybenzoate
2,4-dihydroxybenzoic acid
-
competitive inhibition
2,5-Dihydroxybenzoate
-
-
2-(2H-1,3-benzodioxol-5-yl)-N-[(4-fluorophenyl)methyl]-N-hydroxyacetamide
Q0SH27 AND Q0SH26
60% inhibition at 0.2 mM
2-Fluoro-4-hydroxybenzoate
-
-
2-Hydroxyisonicotinic acid N-oxide
2-Hydroxypyridine N-oxide
-
-
3,4-Dihydroxyacetophenone
3,4-dihydroxybenzoate
-
-
3,4-dihydroxycinnamic acid
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-
3,4-Dihydroxyphenylacetate
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-
3,4-dihydroxyphenylacetic acid
3,4-Dihydroxyphenylpropionate
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-
3,5-Dichloro-4-hydroxybenzoate
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-
3-(3,4-dimethoxyphenyl)-N-hydroxy-N-octylpropanamide
Q0SH27 AND Q0SH26
57% inhibition at 0.2 mM
-
3-(3,4-dimethoxyphenyl)-N-hydroxypropanamide
Q0SH27 AND Q0SH26
60% inhibition at 0.2 mM
3-Bromo-4-hydroxybenzoate
-
-
3-Chloro-4-hydroxybenzoate
-
-
3-Fluoro-4-hydroxybenzoate
3-hydroxybenzoate
3-hydroxyphenylacetic acid
-
-
3-Nitrophenol
-
0.006 mM, 56% inhibition
4-(dimethylamino)benzaldehyde
-
-
4-Fluoro-3-hydroxybenzoate
4-hydroxybenzoate
4-hydroxymercuribenzoate
4-hydroxyphenylacetic acid
-
-
4-methylcatechol
4-nitrocatechol
4-sulfocatechol
alpha-chloro-3,4-dihydroxyacetophenone
-
-
butanol
Ca2+
Nocardia erythropolis
-
-
caffeic acid
-
-
catechol
ethanol
Ferrous ammonium sulfate
-
-
Hg2+
Nocardia erythropolis
-
0.1 mM, 45% inhibition
iodoacetate
isonicotinic acid N-oxide
-
-
Isovanillate
-
-
K+
A0A193DXA9 and A0A193DXP2
16.7% residual activity at 1 mM
KF
-
50 mM, 34% inhibition
methanol
Mg2+
A0A193DXA9 and A0A193DXP2
50% residual activity at 1 mM
N-ethylmaleimide
Na2HAsO4
Nocardia erythropolis
-
0.1 mM, 50% inhibition
Nickel ammonium sulfate
-
-
o-Chloranil
-
0.006 mM, 51% inhibition
p-chloromercuribenzoate
-
0.05 mM, 47% inhibition
Pb2+
Nocardia erythropolis
-
-
Phenanthroline
Propanol
Protocatechualdehyde
protocatechuate
Protocatechuic acid methyl ester
-
-
vanillate
-
-
additional information
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
butanol
EDTA
I0DHJ0 AND I0DHJ1
135.07% activity at 1 mM
ethanol
Na2SO4
-
nonessential activator
Propanol
additional information
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.0029 - 0.15929
3,4-dihydroxybenzoate
0.014
4-Cresol
-
pH 7.5, 38°C
0.014
4-methylcatechol
-
pH 7.5, 38°C
0.042 - 0.058
4-sulfocatechol
0.014
caffeic acid
-
pH 7.5, 38°C
0.014
catechin
-
pH 7.5, 38°C
0.014 - 0.033
catechol
0.04
FeSO4
-
-
0.002
gallate
-
pH 7.5, 38°C
0.014
L-Dopa
-
pH 7.5, 38°C
0.006 - 0.8
O2
0.002 - 0.33
protocatechuate
0.014
pyrogallol
-
pH 7.5, 38°C
additional information
additional information
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.007 - 758
3,4-dihydroxybenzoate
14.6
4-sulfocatechol
-
protocatechuate 3,4-dioxgenase type II
0.007 - 120
O2
0.007 - 100
protocatechuate
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
45.1 - 93.5
3,4-dihydroxybenzoate
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
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 - 0.01
3-Fluoro-4-hydroxybenzoate
0.004
3-hydroxybenzoate
-
-
0.3 - 0.8
4-Fluoro-3-hydroxybenzoate
0.087 - 0.24
4-hydroxybenzoate
0.44
4-methylcatechol
-
-
0.0006 - 0.0048
4-nitrocatechol
0.032 - 0.053
4-sulfocatechol
0.17
alpha-chloro-3,4-dihydroxyacetophenone
-
-
0.38 - 0.7
catechol
1.3
isonicotinic acid N-oxide
-
-
0.02
Isovanillate
-
-
0.001 - 0.073
Protocatechualdehyde
0.066 - 0.102
protocatechuate
0.4
protocatechuic acid methylester
-
-
0.003
vanillate
-
-
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
-
IC50 VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.02
(2E)-3-(3,4-dimethoxyphenyl)-N-hydroxyprop-2-enamide
Rhodococcus jostii
Q0SH27 AND Q0SH26
pH 7.5, 37°C
0.015
(2E)-N-hydroxy-3-(4-methoxyphenyl)prop-2-enamide
Rhodococcus jostii
Q0SH27 AND Q0SH26
pH 7.5, 37°C
0.16
3-(3,4-dimethoxyphenyl)-N-hydroxypropanamide
Rhodococcus jostii
Q0SH27 AND Q0SH26
pH 7.5, 37°C
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.0081
-
-
0.113
-
activity in E. coli cell extracts expressing the recombinant enzyme
0.31
-
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
-
Rosebacter isolate S25com04, enzyme activity in cell extracts
0.88
-
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
37.2
-
recombinant enzyme
105
-
type I protocatechuate 3,4-dioxygenase
121.7
-
purified native enzyme, pH 7.5, 38°C
350
-
activity at the 10th h of the exponential growth phase
additional information
Nocardia erythropolis
-
379 mg O2/min/ml
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5.2
-
completely inactive above pH 6.5
7.3 - 8.5
-
immobilized enzyme
8 - 9.2
I0DHJ0 AND I0DHJ1
-
8.3 - 8.4
-
free enzyme
14
I0DHJ0 AND I0DHJ1
calcium alginate-immobilized enzyme
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
2.2 - 13
I0DHJ0 AND I0DHJ1
glyoxyl agarose-immobilized enzyme, profile overview
4 - 9
-
activity range, profile overview
4 - 14
I0DHJ0 AND I0DHJ1
calcium alginate-immobilized enzyme, profile overview
5.5 - 8.5
A0A193DXA9 and A0A193DXP2
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
6.5 - 8
-
80% of maximal activity within this range
6.5 - 11.5
I0DHJ0 AND I0DHJ1
free enzyme, profile overview
7 - 11
I0DHJ0 AND I0DHJ1
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
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
45
-
immobilized enzyme
45 - 50
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4 - 60
I0DHJ0 AND I0DHJ1
free and immobilized enzyme, profile overview
20 - 70
A0A193DXA9 and A0A193DXP2
the enzyme shows more than 50% activity between 20 and 70°C
45 - 80
-
immobilized enzyme: 78% activity at 60°C, 75% activity at 80°C
50 - 70
-
50°C: about 55% of maximal activity, 70°C: about 60% of maximal activity
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
-
-
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
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
UNIPROT
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
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
21502
-
x * 27097 + x * 21502, deduced from nucleotide sequence
21900
-
x * 21900 + x * 26700, deduced from nucleotide sequences of putative alpha and beta subunits
22000
-
x * 26000 + x * 22000, 2D-PAGE
22280
-
alpha,beta, 2 * 22280 + 2 * 26630, holoenzyme probably contains 6 alpha2,beta2 tetramers, amino acid sequence
22500
-
alpha,beta, 5 * 22500 + 5 * 40000, SDS-PAGE
23300
-
alpha,beta, 5 * 23300 + 5 * 25500, SDS-PAGE
25500
-
alpha,beta, 5 * 23300 + 5 * 25500, SDS-PAGE
26000
-
x * 26000 + x * 22000, 2D-PAGE
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
27097
-
x * 27097 + x * 21502, deduced from nucleotide sequence
34300
-
trimer of dimers, (alphabeta)3, 3 * 29000, alpha subunit, + 3 * 34300, beta-subunit, SDS-PAGE
40000
-
alpha,beta, 5 * 22500 + 5 * 40000, SDS-PAGE
97832
-
8 * 97832, calculated
150000
189900
-
gel filtration
190000
-
gel filtration
202000
-
meniscus depletion
204000
-
sedimentation equilibrium
220000
-
gel filtration
315000
-
analytical ultracentrifugation
480000
500000
A0A193DXA9 and A0A193DXP2
gel filtration
510000
-
gel filtration
590000
-
ultracentrifugation
677000
-
sedimentation velocity analysis
700000
-
diffusion data
783100
-
calculated
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
decamer
heterodimer
hexamer
octamer
tetramer
-
alpha,beta, 2 * 22280 + 2 * 26630, holoenzyme probably contains 6 alpha2,beta2 tetramers, amino acid sequence
additional information
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
no glycoprotein
-
carbohydrate contributes less than 0.2% to the mass of the holoenzyme
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
-
X-ray crystallography
-
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
-
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6 - 9.6
-
24°C, 1 week, less than 10% loss of activity
439483
7.6 - 8.6
Nocardia erythropolis
-
most stable
439502
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5
-
stable for long periods
20 - 50
-
purified enzyme, 50 min, pH 7.5, relatively stable
42
Nocardia erythropolis
-
1 h, stable below
60 - 70
A0A193DXA9 and A0A193DXP2
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
60
-
5 min, stable
65
-
soluble and immobilized enzyme, 15 min, complete loss of activity
70
-
purified enzyme, 20 min, pH 7.5, loss of 95% activity
90
-
the free enzyme loses 82% of relative activity after 180 min of incubations at 90°C
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
enzyme immobilized on porous glass beads is stable over wide ranges of pH and temperature
-
Fe3O4 nanoparticles-immobilized enzyme exhibits improved thermal stability compared to the free enzyme
A0A193DXA9 and A0A193DXP2
immobilization: slight increase of thermal stability, soluble and immobilized enzyme exhibit substantial activity in 2 M urea
-
inactivated by reducing agents
-
lyophilization, stable
-
multi-walled carbon nanotube-immobilized enzyme exhibits improved thermal and pH stabilities compared to the free enzyme
-
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
-
UV irradiation: 38000 ergs per m2, inactivates
-
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-20°C, stable for several years
-
0-3°C, several weeks, no loss in activity
-
25°C, free enzyme, 30 days, 61% loss of activity
-
25°C, immobilized enzyme, 30 days, 56% loss of activity
-
4°C, free enzyme, 30 days, 59% loss of activity
-
4°C, immobilized enzyme, 30 days, 56% loss of activity
-
4°C, pH 8.5, 6 months
-
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
I0DHJ0 AND I0DHJ1
5°C, long periods, no loss in activity
-
stable only 7 days
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
ammonium sulfate, DEAE-cellulose
-
ammonium sulfate, DEAE-cellulose, Bio-gel agarose, DEAE-cellulose
-
ammonium sulfate, DEAE-cellulose, Sephadex G-200
-
ammonium sulfate, DEAE-Sepharose, octyl-Sepharose, Phenyl-sepharose, recombinant enzyme
-
ammonium sulfate, DEAE-Sepharose, Phenyl-Sepharose
-
ammonium sulfate, Q-Sepharose, partial purification
-
ammonium sulfate, Sephacryl S 300, DEAE-Sephadex A50
-
cells harvested by centrifugation, washed twice with 50 mM sodium phosphate buffer, pH 7.0 containing 10% glycerol, disruption with solicitor, centrifuged, supernatant stored at -20°C
-
enzymes from Escherichia coli: DEAE-Sepharose Fast Flow column (5.5 x 19 cm), Phenyl-Sepharose CL-4B column (4.5 x 22.5 cm), Sephacryl S-300 column (3 x 97.5 cm); enzymes from Pseudomonas florescens
-
native enzyme 296.8fold by ammonium sulfate fractionation, anion exchange and hydrophobic interaction chromatography
-
Ni-affinity column chromatography
A0A193DXA9 and A0A193DXP2
Q-Sepharose FF, ammonium sulfate, Sephacryl S-300HR, phenyl-superose HR5/5, Mono Q, type II protocatechuate 3,4-dioxygenase; Q-Sepharose FF, ammonium sulfate, Sephacryl S-300HR, Phenyl-Superose, type I protocatechuate 3,4-dioxygenase
-
Q-Sepharose FF, ammonium sulfate, Superdex 200, Fractogel, type II protocatechuate 3,4-dioxygenase
-
recombinant enzyme
-
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
expressed in Escherichia coli
-
expressed in Escherichia coli BL21(DE3) cells
A0A193DXA9 and A0A193DXP2
expressed in Escherichia coli, the genetic locus ncg12314-ncg12315 of the Corynebacterium glutamicum encodes a putative protocatechuate 3,4-dioxygenase
-
expression in Escherichia coli
-
expression of protocatechuate 3,4-dioxygenase II in Escherichia coli
-
expression of wild-type and Y447H mutant enzyme in Pseudomonas fluorescens
-
expression of wild-type, W153V, R142K and R142/W153V double mutant of protocatechuate 3,4-dioxygenase I in Escherichia coli
-
phylogenetic tree
-
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
-
EXPRESSION
ORGANISM
UNIPROT
LITERATURE
4-hydroxybenzoate induces the protocatechuate 3,4-dioxygenase
I0DHJ0 AND I0DHJ1
4-hydroxybenzoate induces the protocatechuate 3,4-dioxygenase; enzyme expression is induced when cells are grown on 9 mM 4-hydroxybenzoate
-
-
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
enzyme expression is induced when cells are grown on 9 mM 4-hydroxybenzoate
I0DHJ0 AND I0DHJ1
protocatechuate induces PcaHG activity and PcaO regulates the key step in aromatic cleavage during PCA degradation at the pcaHG transcriptional level
-
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
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
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
R153V
-
protocatechuate 3,4-dioxygenase I gain of function mutation, mutant enzyme oxidizes 4-sulfocatechol
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.
additional information
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
analysis
-
use in flushing and scrubbing oxygen out of instruments such as stopped-flow spectrophotometers
degradation
environmental protection
-
the purified enzyme can be used in bioremediation of polluted groundwater or soil contaminated with various aromatic compounds ranging from monocyclic to polycyclic
Show AA Sequence (1310 entries)
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