Information on EC 1.14.13.50 - pentachlorophenol monooxygenase

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The enzyme appears in viruses and cellular organisms

EC NUMBER
COMMENTARY hide
1.14.13.50
-
RECOMMENDED NAME
GeneOntology No.
pentachlorophenol monooxygenase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
2,3,5,6-tetrachlorophenol + NADPH + H+ + O2 = 2,3,5,6-tetrachlorohydroquinone + NADP+ + H2O
show the reaction diagram
-
-
-
-
pentachlorophenol + NADPH + H+ + O2 = 2,3,5,6-tetrachloro-1,4-benzoquinone + NADP+ + chloride + H2O
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
dehalogenation
-
-
-
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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
BRENDA Link
KEGG Link
MetaCyc Link
Chlorocyclohexane and chlorobenzene degradation
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Fluorobenzoate degradation
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Metabolic pathways
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Microbial metabolism in diverse environments
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pentachlorophenol degradation
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SYSTEMATIC NAME
IUBMB Comments
pentachlorophenol,NADPH:oxygen oxidoreductase (hydroxylating, dechlorinating)
A flavoprotein (FAD). The enzyme displaces a diverse range of substituents from the 4-position of polyhalogenated phenols but requires that a halogen substituent be present at the 2-position [2]. The enzyme converts many polyhalogenated phenols into hydroquinones, and requires that a halogen substituent be present at C-2 [2]. If C-4 carries a halogen substituent, reaction 1 is catalysed, e.g. 2,4,6-triiodophenol is oxidized to 2,6-diiodohydroquinone; if C-4 is unsubstituted, reaction 2 is catalysed.
CAS REGISTRY NUMBER
COMMENTARY hide
124148-88-5
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136111-57-4
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ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
strain ATCC 33790
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-
Manually annotated by BRENDA team
i.e. Mycobacterium chlorophenolicus, gene pcpB
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Manually annotated by BRENDA team
strain IST 103, i.e. PCP 103
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Manually annotated by BRENDA team
strain IST 103, i.e. PCP 103
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-
Manually annotated by BRENDA team
UG30
SwissProt
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
metabolism
hydroxylation of pentachlorophenol in the pentachlorophenol biodegradation pathway
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
2,3,4,5-tetrachlorophenol + NADPH + O2
2,3,5-trichlorohydroquinone + Cl- + NADP+ + H2O
show the reaction diagram
-
-
-
-
?
2,3,4,6-tetrachlorophenol + NADPH + O2
?
show the reaction diagram
-
-
-
-
?
2,3,4-trichlorophenol + NADPH + O2
?
show the reaction diagram
2,3,5,6-tetrachlorophenol + 2 NADPH + H+ + O2
2,3,5,6-tetrachloro-p-hydroquinone + 2 NADP+ + chloride + H2O
show the reaction diagram
-
-
-
-
?
2,3,5,6-tetrachlorophenol + NADPH + O2
2,3,5,6-tetrachloro-p-hydroquinone + NADP+
show the reaction diagram
-
-
-
?
2,3,5,6-tetrachlorophenol + NADPH + O2
?
show the reaction diagram
2,3,5-trichlorophenol + NADPH + O2
?
show the reaction diagram
-
-
-
-
?
2,3,6-trichlorophenol + NADPH + O2
2,3,6-trichloro-p-hydroquinone + NADP+ + H2O
show the reaction diagram
-
-
-
?
2,3,6-trichlorophenol + NADPH + O2
?
show the reaction diagram
2,3-dichlorophenol + NADPH + O2
?
show the reaction diagram
-
low activity
-
-
?
2,4,5-trichlorophenol + NADPH + O2
?
show the reaction diagram
-
-
-
-
?
2,4,6-tribromophenol + NADPH + O2
2,6-dibromo-p-hydroquinone + Br- + NADP+ + H2O
show the reaction diagram
-
low activity
-
?
2,4,6-trichlorophenol + NADPH + O2
2,6-dichloro-p-hydroquinone + Cl- + NADP+ + H2O
show the reaction diagram
2,4,6-trichlorophenol + NADPH + O2
?
show the reaction diagram
-
-
-
-
?
2,4-dichlorophenol + NADPH + O2
?
show the reaction diagram
-
-
-
-
?
2,4-dinitrophenol + NADPH + O2
?
show the reaction diagram
-
-
-
-
?
2,6-dibromo-4-nitrophenol + NADPH + O2
2,6-dibromo-p-hydroquinone + NO2- + NADP+ + H2O
show the reaction diagram
-
low activity
-
?
2,6-dibromophenol + NADPH + O2
2,6-dibromo-p-hydroquinone + NADP+ + H2O
show the reaction diagram
-
-
-
?
2,6-dichlorophenol + NADPH + H+ + O2
2,6-dichloro-p-hydroquinone + NADP+ + H2O
show the reaction diagram
-
-
-
-
?
2,6-dichlorophenol + NADPH + O2
2,6-dichloro-p-hydroquinone + NADP+ + H2O
show the reaction diagram
-
-
-
?
2,6-dichlorophenol + NADPH + O2
?
show the reaction diagram
-
-
-
-
?
3,4,5,6-tetrachlorophenol + NADPH + O2
?
show the reaction diagram
-
-
-
-
?
3,4,5-trichlorophenol + 2 NADPH + H+ + O2
3,5-dichloro-p-hydroquinone + 2 NADP+ + chloride + H2O
show the reaction diagram
-
-
-
-
?
3,4,5-trichlorophenol + NADPH + O2
?
show the reaction diagram
-
-
-
-
?
3,5-dibromo-4-hydroxybenzonitrile + 2 NADPH + 2 H+ + O2
2,6-dibromo-p-hydroquinone + CN- + 2 NADP+ + H2O
show the reaction diagram
-
-
-
?
3,5-dibromo-4-hydroxybenzonitrile + NADPH + O2
?
show the reaction diagram
-
-
-
-
?
3,5-dichlorophenol + NADPH + H+ + O2
3,5-dichloro-p-hydroquinone + NADP+ + H2O
show the reaction diagram
-
-
-
-
?
3,5-diiodo-4-hydroxybenzonitrile + NADPH + O2
?
show the reaction diagram
-
-
-
-
?
4,6-dinitro-o-cresol + NADPH + O2
?
show the reaction diagram
-
-
-
-
?
4-amino-2,6-dichlorophenol + NADPH + O2
2,6-dichloro-p-hydroquinone + NADP+ + NH2OH
show the reaction diagram
-
low activity
-
?
4-nitrocatechol + NADPH + O2
?
show the reaction diagram
4-nitrophenol + NADPH + O2
?
show the reaction diagram
-
-
-
-
?
chloroguaiacol + NADPH + O2
?
show the reaction diagram
-
-
-
-
?
p-nitrophenol + NADPH + O2
?
show the reaction diagram
pentachlorophenol + 2 NADPH + H+ + O2
tetrachlorobenzoquinone + 2 NADP+ + chloride + H2O
show the reaction diagram
-
-
-
-
?
pentachlorophenol + 2 NADPH + H+ + O2
tetrachlorohydroquinone + 2 NADP+ + chloride + H2O
show the reaction diagram
-
product is not tetrachlorobenzoquinone as previously thought, ethyl acetate and glutathione can shift the redox-equilibrium in favor of tetrachlorobenzoquinone, room temperature (23-25°C), 50 mM phosphate buffer, pH 7.0, while with NADPH tetrachlorobenzoquinone is reduced to tetrachlorohydroquinone (with 0.5-1 mol NADPH/mol tetrachlorobenzoquinone 50-65% are reduced, with 2.5 mol or more almost complete reduction occurs), deionized distilled water, pH 6.8 (due to interference of phosphate or Tris-HCl buffer with the detection of tetrachlorohydroquinone)
-
?
pentachlorophenol + 2 NADPH + H+ + O2
tetrachlorohydroquinone + 2 NADP+ + Cl- + H2O
show the reaction diagram
pentachlorophenol + NADPH + O2
2,3,5,6-tetrachlorohydroquinone + NADP+ + chloride
show the reaction diagram
pentachlorophenol + NADPH + O2
2,3,5,6-tetrachlorohydroquinone + NADP+ + chloride + H2O
show the reaction diagram
pentachlorophenol + NADPH + O2
tetrachlorobenzoquinone + NADP+ + chloride + H2O
show the reaction diagram
pentachlorophenol + NADPH + O2
tetrachlorohydroquinone + NADP+ + chloride + H2O
show the reaction diagram
pentafluorophenol + NADPH + O2
?
show the reaction diagram
-
low activity
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-
?
tetrachlorobenzoquinone + NADPH
tetrachlorohydroquinone + NADP+
show the reaction diagram
tribromophenol + NADPH + O2
?
show the reaction diagram
-
-
-
-
?
triiodophenol + NADPH + O2
?
show the reaction diagram
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-
-
-
?
triiodophenol + NADPH + O2
diiodohydroquinone + iodide + NADP+ + H2O
show the reaction diagram
Tween 20 + NADPH + O2
?
show the reaction diagram
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-
?
additional information
?
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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
pentachlorophenol + 2 NADPH + H+ + O2
tetrachlorohydroquinone + 2 NADP+ + chloride + H2O
show the reaction diagram
P42535
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product is not tetrachlorobenzoquinone as previously thought, ethyl acetate and glutathione can shift the redox-equilibrium in favor of tetrachlorobenzoquinone, room temperature (23-25°C), 50 mM phosphate buffer, pH 7.0, while with NADPH tetrachlorobenzoquinone is reduced to tetrachlorohydroquinone (with 0.5-1 mol NADPH/mol tetrachlorobenzoquinone 50-65% are reduced, with 2.5 mol or more almost complete reduction occurs), deionized distilled water, pH 6.8 (due to interference of phosphate or Tris-HCl buffer with the detection of tetrachlorohydroquinone)
-
?
pentachlorophenol + 2 NADPH + H+ + O2
tetrachlorohydroquinone + 2 NADP+ + Cl- + H2O
show the reaction diagram
pentachlorophenol + NADPH + O2
2,3,5,6-tetrachlorohydroquinone + NADP+ + chloride + H2O
show the reaction diagram
pentachlorophenol + NADPH + O2
tetrachlorobenzoquinone + NADP+ + chloride + H2O
show the reaction diagram
pentachlorophenol + NADPH + O2
tetrachlorohydroquinone + NADP+ + chloride + H2O
show the reaction diagram
tetrachlorobenzoquinone + NADPH
tetrachlorohydroquinone + NADP+
show the reaction diagram
-
second step in pentachlorophenol degradation, physiological role in association with pentachloro monooxygenase, overview
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?
additional information
?
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COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
NADH
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can replace NADPH, less effective
additional information
-
dithiothreitol can not be used as electron donor
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METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Fe2+
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stimulates
additional information
-
no requirement for divalent cations
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
tetrachlorobenzoquinone
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inactivates the enzyme, feedback control
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
EDTA
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0.01 mM, stimulates by 67%
additional information
-
pentachlorphenol induces enzyme expression
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KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.0056
2,3,5,6-Tetrachlorophenol
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pH 7.0, 22°C
0.0126
2,6-dichlorophenol
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pH 7.0, 22°C
0.059
3,4,5-trichlorophenol
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pH 7.0, 22°C
0.098
3,5-dichlorophenol
-
pH 7.0, 22°C
0.214
NADH
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-
0.082
NADPH
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-
0.0005 - 0.05
Pentachlorophenol
additional information
additional information
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TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.32
2,3,5,6-Tetrachlorophenol
Sphingobium chlorophenolicum
-
pH 7.0, 22°C
0.046
2,6-dichlorophenol
Sphingobium chlorophenolicum
-
pH 7.0, 22°C
0.01
3,4,5-trichlorophenol
Sphingobium chlorophenolicum
-
pH 7.0, 22°C
0.01
3,5-dichlorophenol
Sphingobium chlorophenolicum
-
pH 7.0, 22°C
0.024 - 16000
Pentachlorophenol
additional information
additional information
Sphingobium chlorophenolicum
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kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
57
2,3,5,6-Tetrachlorophenol
Sphingobium chlorophenolicum
-
pH 7.0, 22°C
6415
3.81
2,6-dichlorophenol
Sphingobium chlorophenolicum
-
pH 7.0, 22°C
5300
0.169
3,4,5-trichlorophenol
Sphingobium chlorophenolicum
-
pH 7.0, 22°C
24578
0.102
3,5-dichlorophenol
Sphingobium chlorophenolicum
-
pH 7.0, 22°C
11176
24
Pentachlorophenol
Sphingobium chlorophenolicum
-
pH 7.0, 22°C
1077
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
3.7
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purified enzyme
additional information
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
7.5 - 8.5
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-
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6 - 8.5
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pH 6.0: about 28% of activity maximum, pH 7.5-8.5: activity maximum
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
40 - 45
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40°C: activity maximum, 45°C: above 75% of activity maximum, 50°C: inactive
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
additional information
-
isolate from contaminated site
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
-
enzyme is synthesized in the cytoplasm, then integrated in the inner membranes and finally translocated to the periplasm
Manually annotated by BRENDA team
-
enzyme is synthesized in the cytoplasm, then integrated in the inner membranes and finally translocated to the periplasm
Manually annotated by BRENDA team
-
enzyme is synthesized in the cytoplasm, then integrated in the inner membranes and finally translocated to the periplasm
-
Manually annotated by BRENDA team
additional information
-
probably not a soluble protein
-
Manually annotated by BRENDA team
PDB
SCOP
CATH
ORGANISM
UNIPROT
Nocardia farcinica (strain IFM 10152)
Nocardia farcinica (strain IFM 10152)
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
24000
-
gel filtration
30000
-
gel filtration
59930
deduced from DNA sequence determination
60000
-
x * 60000, SDS-PAGE
63000
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1 * 63000, SDS-PAGE, under certain conditions dimer and multimer conformations are also observed
66000
-
gel filtration
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
monomer
additional information
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-20°C, as ammonium sulfate precipitate or in solution at concentration greater than 1 mg/ml, 2 months
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4°C, 50% loss of activity after 1 week
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4°C, in cell extract or as 40-60% fraction of ammonium sulfate precipitation, total loss of activity after 3 days
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
from culture extract to homogeneity, about 13fold
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frozen pellet of cell culture is suspended in lysis buffer (50 mM Tris-HCl, pH 7.7, 025 mM phenylmethanesulfonyl fluoride, 1 microM pepstatin A, lysozyme), cells disrupted by sonication, centrifuged, supernatant purified by affinity chromatography, mixed with Ni-NTA agarose medium, washed, packed into a column, eluted, active fractions combined and stored in buffer (20 mM phosphate buffer, pH 7.0, 0.25 mM phenylmethanesulfonyl fluoride, 5% glycerol)
partially
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recombinant enzyme from Escherichia coli to homogeneity; recombinant enzyme from Pseudomonas aeruginosa to over 95% homogeneity by nickel affinity chromatography
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recombinant wild-type and mutant enzymes from Escherichia coli
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recombinant wild-type and mutant enzymes from Escherichia coli strain Bl21(DE3)
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recombinant wild-type and mutant from Escherichia coli as glutathione-S-transferase fusion proteins
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Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
DNA and amino acid sequence determination and analysis, expression of His-tagged wild-type and mutant enzymes in Escherichia coli
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Escherichia coli M15 overexpressing recombinant hexahistidine-tagged pentachlorophenol 4-monooxygenase
expression in Escherichia coli
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expression of wild-type and mutant enzymes in Escherichia coli strain BL21(DE3)
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gene isolated from Pseudomonas sp. plasmid, recombinant expression in Escherichia coli XLBlue1alpha, DNA sequence analysis of the recombinant clones, comparison with other sequences
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gene pcpB, DNA and amino acid sequence analysis, phylogenetic analysis
gene pcpB, expression in Escherichia coli; gene pcpD, expression as C-terminally His12-tagged protein in Pseudomonas aeruginosa strain PAO1-Lac
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pcpB gene, constitutive overexpression in Escherichia coli, DNA sequence determination and analysis
pcpB gene, expression in Escherichia coli and Flavobacterium sp. ATCC 39723 of wild-type and isogenic mutant, DNA sequence analysis
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pcpB gene, expression in Escherichia coli BL21, DNA sequence analysis
pcpB gene, expression of wild-type and mutant in Escherichia coli as glutathione-S-transferase fusion protein
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ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
F223A
-
site-directed mutagenesis, increased activity compared to the wild-type enzyme
F225A
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site-directed mutagenesis, increased activity compared to the wild-type enzyme
F87A
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site-directed mutagenesis, mutant enzyme is nearly not soluble in contrast to the wild-type, highly reduced activity compared to the wild-type enzyme
moe
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construction of an allelic exchange mutant and a pcpD knockout mutant strains, the latter show reduced activity at high pentachlorophenol concentrations, not at low concentrations
R235A
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site-directed mutagenesis, determination of molecular weight and comparison to wild-type enzyme
Y216F
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site-directed mutagenesis, 60% decreased activity compared to the wild-type enzyme
additional information
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
degradation
pentachlorophenol is a chloroaromatic pesticide used to protect lumber, and an environmental pollutant, Sphingobium chlorophenolicum is a microorganism that can degrade the agent to 3-oxoadipate using 5 catalytic enzymes, pentachlorophenol 4-monooxygenase catalyzes the first and rate-limiting step
environmental protection
additional information
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