Information on EC 1.14.12.18 - biphenyl 2,3-dioxygenase

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The enzyme appears in viruses and cellular organisms

EC NUMBER
COMMENTARY hide
1.14.12.18
-
RECOMMENDED NAME
GeneOntology No.
biphenyl 2,3-dioxygenase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
biphenyl + NADH + H+ + O2 = (1S,2R)-3-phenylcyclohexa-3,5-diene-1,2-diol + NAD+
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
oxidation
-
-
-
-
redox reaction
reduction
-
-
-
-
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
biphenyl degradation
-
-
Dioxin degradation
-
-
diphenyl ethers degradation
-
-
Metabolic pathways
-
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Microbial metabolism in diverse environments
-
-
SYSTEMATIC NAME
IUBMB Comments
biphenyl,NADH:oxygen oxidoreductase (2,3-hydroxylating)
Requires Fe2+. The enzyme from Burkholderia fungorum LB400 (previously Pseudomonas sp.) is part of a multicomponent system composed of an NADH:ferredoxin oxidoreductase (FAD cofactor), a [2Fe-2S] Rieske-type ferredoxin, and a terminal oxygenase that contains a [2Fe-2S] Rieske-type iron-sulfur cluster and a catalytic mononuclear nonheme iron centre. Chlorine-substituted biphenyls can also act as substrates. Similar to the three-component enzyme systems EC 1.14.12.3 (benzene 1,2-dioxygenase) and EC 1.14.12.11 (toluene dioxygenase).
CAS REGISTRY NUMBER
COMMENTARY hide
103289-55-0
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
-
-
-
Manually annotated by BRENDA team
large subunit
UniProt
Manually annotated by BRENDA team
large subunit fragment; strain B-358
UniProt
Manually annotated by BRENDA team
large subunit fragment; strain YT01
UniProt
Manually annotated by BRENDA team
strain LB4000
-
-
Manually annotated by BRENDA team
strain A5
-
-
Manually annotated by BRENDA team
strain A5
-
-
Manually annotated by BRENDA team
isolated from polluted soil
-
-
Manually annotated by BRENDA team
isolated from polluted soil
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
large subunit fragment
UniProt
Manually annotated by BRENDA team
strain B-356
-
-
Manually annotated by BRENDA team
strain B-356
-
-
Manually annotated by BRENDA team
strain JI104
-
-
Manually annotated by BRENDA team
strain JI104
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
Cam1
-
-
Manually annotated by BRENDA team
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-
-
Manually annotated by BRENDA team
strain R04
-
-
Manually annotated by BRENDA team
strain B8/36, former name Beijerinckia sp.
-
-
Manually annotated by BRENDA team
large subunit fragment; uncultured soil bacterium
UniProt
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
metabolism
physiological function
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
(3R,4R)-cis-isoflavan-4-ol + NADH + H+ + O2
(3R,4R)-3-(7-oxabicyclo[4.1.0]hepta-1,3,5-trien-2-yl)-3,4-dihydro-2H-chromen-4-ol + NAD+
show the reaction diagram
(3R,4S)-trans-isoflavan-4-ol + NADH + H+ + O2
(3R,4S)-3-(7-oxabicyclo[4.1.0]hepta-1,3,5-trien-2-yl)-3,4-dihydro-2H-chromen-4-ol + NAD+
show the reaction diagram
(3S,4R)-trans-isoflavan-4-ol + NADH + H+ + O2
(3S,4R)-3-(7-oxabicyclo[4.1.0]hepta-1,3,5-trien-2-yl)-3,4-dihydro-2H-chromen-4-ol + NAD+
show the reaction diagram
(3S,4S)-cis-isoflavan-4-ol + NADH + H+ + O2
(3S,4S)-3-(7-oxabicyclo[4.1.0]hepta-1,3,5-trien-2-yl)-3,4-dihydro-2H-chromen-4-ol + NAD+
show the reaction diagram
-
-
-
-
?
2,2',3,3'-tetrachlorobiphenyl + NADH + H+ + O2
5,6-dihydro-5,6-dihydroxy-2,2',3,3'-tetrachlorobiphenyl + 4,5-dihydro-4,5-dihydroxy-2,2',3,3'-tetrachlorobiphenyl + NAD+
show the reaction diagram
-
-
-
-
?
2,2',3,3'-tetrachlorobiphenyl + NADH + H+ + O2
?
show the reaction diagram
2,2',3,3'-tetrachlorobiphenyl + NADH + O2
?
show the reaction diagram
2,2',5,5'-tetrachlorobiphenyl + NADH + H+ + O2
3,4-dihydro-3,4-dihydroxy-2,2',5,5'-tetrachlorobiphenyl + NAD+
show the reaction diagram
-
-
-
-
?
2,2',5,5'-tetrachlorobiphenyl + NADH + H+ + O2
?
show the reaction diagram
2,2',5,5'-tetrachlorobiphenyl + NADH + O2
?
show the reaction diagram
2,2'-dibromobiphenyl + NADH + H+ + O2
?
show the reaction diagram
2,2'-dibromobiphenyl + NADH + O2
2,3-dihydroxy-2'-bromobiphenyl + NAD+ + HBr
show the reaction diagram
2,2'-dichlorobiphenyl + NADH + H+ + O2
2,3-dihydroxy-2'-chlorobiphenyl + 3,4-dihydro-3,4-dihydroxy-2,2'-dichlorobiphenyl + NAD+
show the reaction diagram
2,2'-dichlorobiphenyl + NADH + H+ + O2
2,3-dihydroxy-2'-chlorobiphenyl + cis-3,4-dihydro-3,4-dihydroxy-2,2'-dichlorobiphenyl + NAD+ + HCl
show the reaction diagram
-
-
-
-
?
2,2'-dichlorobiphenyl + NADH + H+ + O2
2,3-dihydroxy-2'-chlorobiphenyl + NAD+
show the reaction diagram
-
poor substrate
-
-
?
2,2'-dichlorobiphenyl + NADH + H+ + O2
?
show the reaction diagram
2,2'-dichlorobiphenyl + NADH + O2
2,3-dihydroxy-2'-chlorobiphenyl + 3,4-dihydro-3,4-dihydroxy-2,2'-dichlorobiphenyl + NAD+
show the reaction diagram
2,2'-dichlorobiphenyl + NADH + O2
2,3-dihydroxy-2'-chlorobiphenyl + NAD+ + HCl
show the reaction diagram
2,2'-dichlorobiphenyl + NADH + O2
5,6-dihydroxy-1-phenylcyclohexa-1,3-diene + 2,3-dihydroxy-2'-chlorobiphenyl + NAD+ + HCl
show the reaction diagram
2,2'-difluorobiphenyl + NADH + H+ + O2
?
show the reaction diagram
2,2'-difluorobiphenyl + NADH + O2
2,3-dihydroxy-2'-fluorobiphenyl + 5,6-dihydroxy-2,2'-difluorobiphenyl + NAD+ + HF
show the reaction diagram
2,2'-dihydroxybiphenyl + NADH + H+ + O2
?
show the reaction diagram
2,2'-dihydroxybiphenyl + NADH + O2
2,3,2'-trihydroxybiphenyl + trihydroxybiphenyl + NAD+
show the reaction diagram
2,2'-dinitrobiphenyl + NADH + H+ + O2
?
show the reaction diagram
-
-
-
-
?
2,2'-dinitrobiphenyl + NADH + O2
2,3-dihydroxy-2'-nitrobiphenyl + NAD+ + NO2
show the reaction diagram
2,3',4'-trichlorobiphenyl + NADH + O2
?
show the reaction diagram
-
-
-
-
?
2,3',4-trichlorobiphenyl + NADH + H+ + O2
?
show the reaction diagram
2,3'-dichlorobiphenyl + NADH + H+ + O2
3-chloro-5-(2-chlorophenyl)cyclohexa-3,5-diene-1,2-diol + 4-chloro-3-(3-chlorophenyl)cyclohexa-3,5-diene-1,2-diol + 5-chloro-3-(2-chlorophenyl)cyclohexa-3,5-diene-1,2-diol
show the reaction diagram
2,3'-dichlorobiphenyl + NADH + O2
5,6-dihydroxy-1-phenylcyclohexa-1,3-diene + 2,3-dihydroxy-3'-chlorobiphenyl + 5',6'-dihydroxy-1'-phenylcyclohexa-1',3'-diene + NAD+ + HCl
show the reaction diagram
-
-
-
-
?
2,3,2',3'-tetrachlorobiphenyl + NADH + H+ + O2
4,5-dihydro-4,5-dihydroxy-2,3,2',3'-tetrachlorobiphenyl + NAD+
show the reaction diagram
-
oxygenated onto meta-para carbons 4 and 5
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-
?
2,3,2',3'-tetrachlorobiphenyl + NADH + H+ + O2
cis-4,5-dihydro-4,5-dihydroxy-2,3,2',3'-tetrachlorobiphenyl + NAD+
show the reaction diagram
-
-
-
-
?
2,3,4'-trichlorobiphenyl + NADH + H+ + O2
?
show the reaction diagram
2,3,4'-trichlorobiphenyl + NADH + O2
?
show the reaction diagram
-
-
-
-
?
2,4',5-trichlorobiphenyl + NADH + H+ + O2
3,4-dihydro-3,4-dihydroxy-2,5,4'-trichlorobiphenyl + 2',3'-dihydro-2',3'-dihydroxy-2,5,4'-trichlorobiphenyl + NAD+
show the reaction diagram
-
-
-
-
?
2,4'-dichlorobiphenyl + NADH + O2
2,3-dihydroxy-4'-chlorobiphenyl + NAD+ + HCl
show the reaction diagram
2,4,2',4'-tetrachlorobiphenyl + NADH + H+ + O2
2,3-dihydroxy-2',4,4'-trichlorobiphenyl + NAD+ + HCl
show the reaction diagram
-
oxygenated principally onto vicinal ortho-meta carbons 2 and 3
-
-
?
2,4,4'-trichlorobiphenyl + NADH + H+ + O2
2,3-dihydro-2,3-dihydroxy-2',4,4'-trichlorobiphenyl + NAD+
show the reaction diagram
2,5,2',5'-tetrachlorobiphenyl + NADH + O2
cis-3,4-dihydroxy-2,5-dichloro-1-[2',5'-dichlorophenyl]-cyclohexa-1,5-diene + 3,4-dihydroxy-1-phenylcyclohexa-1,5-diene + NAD+ + HCl
show the reaction diagram
2,5,2'-trichlorobiphenyl + NADH + O2
cis-3,4-dihydroxy-2,5-dichloro-1-[2'-chlorophenyl]-cyclohexa-1,5-diene + 2',3'-dihydroxy-2,5-dichlorobiphenyl + 5',6'-dihydrodiol + NAD+ + HCl
show the reaction diagram
-
-
-
-
?
2,5,3'-trichlorobiphenyl + NADH + O2
3,4-dihydroxy-1-phenylcyclohexa-1,5-diene + 5',6'-dihydroxy-1'-phenylcyclohexa-1',3'-diene + 2,3-catechol + NAD+ + HCl
show the reaction diagram
-
-
-
-
?
2,5,4'-trichlorobiphenyl + NADH + O2
?
show the reaction diagram
2,5-dichlorobiphenyl + NADH + O2
cis-2',3'-dihydroxy-1'-(2,5-dichlorophenyl)-cyclohexa-4',6'-diene + 3,4-dihydroxy-1-phenylcyclohexa-1,5-diene + NAD+ + HCl
show the reaction diagram
2,6',4'-trichlorobiphenyl + NADH + O2
?
show the reaction diagram
-
-
-
-
?
2,6-dichlorobiphenyl + NADH + H+ + O2
2,6-dichloro-2',3'-dihydro-2',3'-dihydroxybiphenyl + ?
show the reaction diagram
2,6-dichlorobiphenyl + NADH + H+ + O2
?
show the reaction diagram
-
poor substrate
-
-
?
2-chlorobiphenyl + NADH + O2
cis-2',3'-dihydroxy-1'-(2-chlorophenyl)-cyclohexa-4',6'-diene + catechol + NAD+
show the reaction diagram
2-hydroxy-3,5-dichlorobiphenyl + NADH + H+ + O2
?
show the reaction diagram
2-hydroxy-3-chlorobiphenyl + NADH + H+ + O2
5-(3-chloro-2-hydroxyphenyl)-6-hydroxy-3-cyclohexen-1-one + NAD+
show the reaction diagram
2-hydroxy-5-chlorobiphenyl + NADH + H+ + O2
cis-2,3-dihydro-2,3-dihydroxy-2'-hydroxy-5'-chlorobiphenyl + 5-(5-chloro-2-hydroxyphenyl)-6-hydroxy-3-cyclohexene-1-one + 2,2'-dihydroxy-5-chlorobiphenyl + 2,3'-dihydroxy-5-chlorobiphenyl + NAD+
show the reaction diagram
3,3'-dichlorobiphenyl + NADH + H+ + O2
5,6-dihydroxy-1-phenylcyclohexa-1,3-diene + 4,5-dihydroxy-1-phenylcyclohexa-1,2-diene + NAD+ + HCl
show the reaction diagram
-
-
-
?
3,3'-dichlorobiphenyl + NADH + H+ + O2
?
show the reaction diagram
3,3'-dichlorobiphenyl + NADH + O2
5,6-dihydroxy-1-phenylcyclohexa-1,3-diene + 4,5-dihydroxy-1-phenylcyclohexa-1,2-diene + NAD+ + HCl
show the reaction diagram
3,3'-dichlorobiphenyl + NADH + O2
?
show the reaction diagram
-
-
-
-
?
3,4'-dichlorobiphenyl + NAD(P)H + O2
5,6-dihydroxy-3,4'-dichlorobiphenyl + NAD(P)+
show the reaction diagram
3,4'-dichlorobiphenyl + NADH + H+ + O2
3-chloro-5-(4-chlorophenyl)cyclohexa-3,5-diene-1,2-diol + 3-chloro-6-(3-chlorophenyl)cyclohexa-3,4-diene-1,2-diol
show the reaction diagram
3,4,4'-trichlorobiphenyl + NADH + O2
?
show the reaction diagram
-
-
-
-
?
3,5',4'-trichlorobiphenyl + NADH + O2
?
show the reaction diagram
-
-
-
-
?
3-chlorobiphenyl + NADH + O2
cis-2',3'-dihydroxy-1'-(3-chlorophenyl)-cyclohexa-4',6'-diene + NAD+
show the reaction diagram
4,4'-dichlorobiphenyl + NAD(P)H + O2
2,3-dihydroxy-4,4'-dichlorobiphenyl + NAD(P)+
show the reaction diagram
4,4'-dichlorobiphenyl + NADH + H+ + O2
2,3-dihydroxy-4,4'-dichlorobiphenyl + NAD+
show the reaction diagram
-
-
-
?
4,4'-dichlorobiphenyl + NADH + H+ + O2
?
show the reaction diagram
4-chlorobiphenyl + NADH + O2
2',3'-dihydrodiol-4-chlorobiphenyl + NAD+
show the reaction diagram
4-methylbiphenyl + NADH + O2
2-hydroxy-6-oxo-6-[4-methylphenyl]-hexa-2,4-dienoic acid + NAD+
show the reaction diagram
5,7-dihydroxyflavone + NADH + H+ + O2
2-(2,3-dihydroxyphenyl)-5,7-dihydroxy-chromen-4-one + NAD+
show the reaction diagram
6,7-dihydro-5H-benzocycloheptene + NAD(P)H + O2
(-)-cis-(1R,2S)-dihydroxybenzocycloheptane + NAD(P)+
show the reaction diagram
6-hydroxyflavanone + O2 + NAD(P)H
?
show the reaction diagram
7-hydroxyflavanone + O2 + NAD(P)H
?
show the reaction diagram
7-hydroxyflavone + NADH + H+ + O2
2-(2,3-dihydroxyphenyl)-7-hydroxy-chromen-4-one + NAD+
show the reaction diagram
benzene + ?
?
show the reaction diagram
-
benzene is only a substrate for Pseudomonas pseudoalcaligenes strain 1072
-
-
?
benzo[a]pyrene + NADH + O2
?
show the reaction diagram
biphenyl + NAD(P)H + O2
(1S,2R)-dihydroxy-3-phenylcyclohexa-3,5-diene + NAD(P)+
show the reaction diagram
biphenyl + NAD(P)H + O2
2,3-dihydro-dihydroxybiphenyl + NAD(P)+
show the reaction diagram
biphenyl + NAD(P)H + O2
cis-biphenyl 2,3-dihydrodiol + NAD(P)+
show the reaction diagram
biphenyl + NADH + H+ + O2
(1S,2R)-3-phenylcyclohexa-3,5-diene-1,2-diol + NAD+
show the reaction diagram
biphenyl + NADH + H+ + O2
(1S,2R)-3-phenylcyclohexa-3,5-diene-1,2-diol + NAD+ + ?
show the reaction diagram
-
-
-
-
?
biphenyl + NADH + H+ + O2
2,3-dihydro-dihydroxybiphenyl + NAD+
show the reaction diagram
biphenyl + NADH + H+ + O2
?
show the reaction diagram
biphenyl + NADH + H+ + O2
cis-(2R,3S)-dihydroxy-1-phenylcyclohexa-4,6-diene + NAD+
show the reaction diagram
-
-
-
-
?
biphenyl + NADH + O2
cis-biphenyl 2,3-dihydrodiol + NAD+
show the reaction diagram
carbazole + NADH + O2
3-hydroxycarbazole + NAD+
show the reaction diagram
dibenzo-p-dioxin + NADH + O2
2,2',3-trihydroxybiphenyl ether + NAD+
show the reaction diagram
dibenzo-p-dioxine + NADH + O2
2,3,2'-trihydroxy-diphenylether + dibenzo-p-dioxine-dihydrodiol + NAD+
show the reaction diagram
-
-
-
-
?
dibenzofuran + NADH + O2
?
show the reaction diagram
-
dibenzofuran is a poor substrate
-
-
?
dibenzofurane + NADH + O2
2,2',3-trihydroxybiphenyl + dihydro-dihydroxy-dibenzofuran + NAD+
show the reaction diagram
dibenzofurane + NADH + O2
monohydroxydibenzofuran + 2,3,2'-trihydroxybiphenyl + dibenzofuran-1,2-dihydrodiol + dibenzofuran-3,4-dihydrodiol + NAD+
show the reaction diagram
-
-
-
-
?
diphenylmethane + NADH + O2
?
show the reaction diagram
-
-
-
-
?
ethylbenzene + ?
?
show the reaction diagram
-
-
-
-
?
ethylbenzene + NADH + O2
?
show the reaction diagram
-
-
-
-
?
flavone + O2 + NAD(P)H
cis-flavone-2',3'-dihydrodiol + ?
show the reaction diagram
-
-
-
-
?
flavone + O2 + NADH + H+
cis-flavone-2',3'-dihydrodiol + ?
show the reaction diagram
isoflavone + NADH + H+ + O2
isoflavone-cis-(2'R,3'S)-dihydrodiol + NAD+
show the reaction diagram
-
-
-
-
?
isoflavone + O2 + NADH + H+
2',3'-dihydro-2',3'-cis-dihydroxyflavone + NAD+
show the reaction diagram
naphthalene + NADH + O2
?
show the reaction diagram
naphthalene + NADPH + O2
cis-naphthalene 1,2-dihydrodiol + NADP+
show the reaction diagram
phenanthrene + NADH + O2
?
show the reaction diagram
toluene + NAD(P)H + O2
?
show the reaction diagram
-
toluene is only a substrate for Pseudomonas pseudoalcaligenes strain 1072
-
-
?
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
biphenyl + NADH + H+ + O2
(1S,2R)-3-phenylcyclohexa-3,5-diene-1,2-diol + NAD+
show the reaction diagram
biphenyl + NADH + H+ + O2
2,3-dihydro-dihydroxybiphenyl + NAD+
show the reaction diagram
biphenyl + NADH + H+ + O2
cis-(2R,3S)-dihydroxy-1-phenylcyclohexa-4,6-diene + NAD+
show the reaction diagram
-
-
-
-
?
additional information
?
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Iron
-
exogenous ferrous iron is required for maximal activity, the oxygenase component contains 2.43 g/mol iron, the ferredoxin component contains 2.1 g/mol iron
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
additional information
-
exposure of soil microcosms to different polychlorinated biphenyl congeners results in different bacterial community structures and abundance of BPH genes
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.0026
2,2'-dichlorobiphenyl
-
recombinant, purified protein
0.0033
3,3'-dichlorobiphenyl
-
recombinant, purified protein
0.00433
Benzene
-
Pseudomonas pseudoalcaligenes strain 1072
0.00018 - 1.1
Biphenyl
0.0108 - 0.0236
ethylbenzene
0.058
NADH
-
recombinant, purified NADH: ferredoxin oxidoreductase component
0.156
NADPH
-
recombinant, purified NADH: ferredoxin oxidoreductase component
0.028
O2
-
recombinant, purified protein
0.0117
Toluene
-
Pseudomonas pseudoalcaligenes strain 1072
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.056
Benzene
Pseudomonas pseudoalcaligenes
-
Pseudomonas pseudoalcaligenes strain 1072
0.113 - 4.6
Biphenyl
0.0497 - 0.219
ethylbenzene
0.115
Toluene
Pseudomonas pseudoalcaligenes
-
Pseudomonas pseudoalcaligenes strain 1072
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.15
-
referred to iron sulfur protein (terminal oxygenase) after purification
4.9
-
recombinant protein after purification
28.9
-
oxygenase component, crude cell extract
217
-
oxygenase component, after 10fold purification
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
7
-
assay at
7.2
-
NADH: ferredoxin oxidoreductase component
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
30
-
assay at
32
-
NADH: ferredoxin oxidoreductase component
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
additional information
PDB
SCOP
CATH
ORGANISM
UNIPROT
Paraburkholderia xenovorans (strain LB400)
Paraburkholderia xenovorans (strain LB400)
Paraburkholderia xenovorans (strain LB400)
Paraburkholderia xenovorans (strain LB400)
Paraburkholderia xenovorans (strain LB400)
Paraburkholderia xenovorans (strain LB400)
Paraburkholderia xenovorans (strain LB400)
Paraburkholderia xenovorans (strain LB400)
Paraburkholderia xenovorans (strain LB400)
Rhodococcus jostii (strain RHA1)
Rhodococcus jostii (strain RHA1)
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
12000
-
ferredoxin component, SDS-PAGE
25200
-
beta subunit with His-tag, SDS-PAGE
27300
-
beta subunit of ISP, SDS-PAGE
41500
-
NADH: ferredoxin component, gel filtration with Superose 6
43000
-
reductase component, SDS-PAGE
43600
-
NADH: ferredoxin component, SDS-PAGE
44000
-
native, recombinant and purified alpha subunit with His-tag, HPLC gel filtration
53000
-
alpha subunit of ISP, SDS-PAGE
53600
-
alpha subunit with His-tag, SDS-PAGE
186000
-
native protein with His-tag, HPLC gel filtration
209000
-
native protein, gel filtration with Superose 12
217000
-
oxygenase component, SDS-PAGE
234000
-
native protein, HPLC gel filtration
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
heterohexamer
heterotetramer
heterotrimer
additional information
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
crystal structure of the BPDOB356–2,6-dichlorobiphenyl complex is determined at 2.4 A resolution
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crystal structure of the BPDOB356/2,6-dichlorobiphenyl complex is determined at 2.4 A resolution
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purified recombinant BPDOLB400, sitting drop vapour diffusion method, anaerobic conditions, method optimization, 0.002 ml of 8 mg/ml protein in 25 mM HEPES, pH 7.3, containing 0.25 mM ferrous ammonium sulfate, and 2 mM dithiothreitol, is mixed with 0.002 ml of reservoir solution containing 20-25% w/v PEG 8000 or PEG 5000 MME, 50 mM PIPES, pH 6.5, 100 mM ammonium acetate, 5% v/v glycerol, and 0.2% w/v agarose, successful crystallization only occurs at pH 6.5, X-ray diffraction structure determination and analysis at 2.4-2.8 A resolution; purified recombinant BPDOLB400, sitting drop vapour diffusion method, anaerobic conditions, method optimization, 0.002 ml of 8 mg/ml protein in 25 mM HEPES, pH 7.3, containing 0.25 mM ferrous ammonium sulfate, and 2 mM dithiothreitol, is mixed with 0.002 ml of reservoir solution containing 20-25% w/v PEG 8000 or PEG 5000 MME, 50 mM PIPES, pH 6.5, 100 mM ammonium acetate, 5% v/v glycerol, and 0.2% w/v agarose, successful crystallization only occurs at pH 6.5, X-ray diffraction structure determination and analysis at 2.4-2.8 A resolution; sitting drop vapor diffusion method, using 20-25% (w/v) PEG 8000 or PEG 5000 MME, 50 mM PIPES pH 6.5, 100 mM ammonium acetate, 5% (v/v) glycerol and 0.2% (w/v) agarose
sitting drop vapor diffusion method, using 20–25% (w/v) PEG 8000 or PEG 5000 MME, 50 mM PIPES (pH 6.5), 100 mM ammonium acetate, 5% (v/v) glycerol, and 0.2% (w/v) agarose
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terminal oxygenase component of the biphenyl dioxygenase, BphA1A2 in substrate-free and biphenyl complex forms, cocrystallization method
hanging drop vapour diffusion method using 20% (w/v) polyethylene glycol 3350, 1 M NaCl and 0.1 M ZnCl2 at 6°C
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TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5 - 55
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NADH: ferredoxin oxidoreductase component: no activity at 5 or 55°C, 15.2% of control activity at 50°C
57
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the oxygenase component has a half-life of 16 min
OXIDATION STABILITY
ORGANISM
UNIPROT
LITERATURE
20 min preincubation of His-tagged enzyme with 5 mM dithiothreitol on ice can restore activity of older preparations
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285287
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-70°C, several months
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-75°C, 4 months, 10% loss of activity
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Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
; recombinant His-tagged subunits alpha and beta and mutant variants from Escherichia coli strain C41 (DE3) by affinity chromatography, the tag is cleaved off by thrombin; recombinant His-tagged subunits alpha and beta and mutant variants from Escherichia coli strain C41(DE3) by affinity chromatography, the tag is cleaved off by thrombin
active BphAE and BphG copurified from Nicotiana benthamiana leaves
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butyl-Sepharose XK26/40 column chromatography, Q-Sepharose column chromatography, and hydroxyapatite column chromatography
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His-tagged ISP, expressed in Escherichia coli
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His-tagged ISP, expressed in Escherichia coli and Pseudomonas putida
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purification from heterologous expression in Pseudomonas putida KT2442
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purification of the iron sulfur protein (ISP) of biphenyl 2,3-dioxygenase
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purification of the NADH: ferredoxin oxidoreductase component
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Q-Sepharose FF column chromatography
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
A central part (amino acid position 268-397 of 458 residues) of the biphenyl dioxygenase large alpha subunit, BphA1, from Pseudomonas pseudoalcaligenes (strain KF707) is exchanged with the corresponding part of BphA1 from Pseudomonas putida strain KF715, to construct hybrid BphA1, BphA1 (amino acid 715-707). When expressed in Escherichia coli together with the bphA2A3A4BC genes from strain KF707, this enzyme is shown to possess activity for degrading both 1-phenylnaphthalene and 2-phenylnaphthalene.
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based on analyses with Nicotiana benthamiana plants transiently expressing the biphenyl dioxygenase genes (the two subunit oxygenase (BphAE) containing a Rieske-type iron–sulfur cluster and a mononuclear iron center, the Rieske-type ferredoxin (BphF), and the FAD-containing ferredoxin reductase (BphG)) from Burkholderia xenovorans LB400 and transgenic Nicotiana tabacum plants transformed with each of these four genes, it can be shown that each of the three biphenyl dioxygenase components can be produced individually as active protein in tobacco plants
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expressed in Escherichia coli
expressed in Escherichia coli C41 (DE3) cells; expression of His-tagged subunits alpha and beta and mutant variants in Escherichia coli strain C41 (DE3); expression of His-tagged subunits alpha and beta and mutant variants in Escherichia coli strain C41(DE3)
expressed in Escherichia coli DH11S cells
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expressed in Escherichia coli JM109 (pJHF108)
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expressed in Escherichia coli JM109 cells
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expressed in Escherichia coli strain BL21Star
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expressed in Escherichia coli Top10F cells
expression in Escherichia coli
gene cluster bphAaAbAcAd, genetic organization, the multicomponent enzyme components cannot be expressed in Escherichia coli actively due to lack of activity of the ferredoxin component in Escherichia coli involving codon usage bias. Therefore hybrid BphA gene derivatives are constructed by replacing ferredoxin and/or reductase components of RHA1 with those of Pseudomonas pseudoalcaligenes strain KF707. Expression of ferredoxin encoding gene bphAc in Escherichia coli strain Rosetta (DE3) pLacI resulting in an active protein
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genes bphAaAbAcAd, encoding the large and small subunits of the terminal oxygenase component and the ferredoxin and reductase subunits responsible for electron transfer from NADH to the large subunit. Functional expression Burkholderia sp. strain NK8 is only successful by combining the bphAaAbAcAd genes of RHA1 and bphA3A4 of Pseudomonas pseudoalcaligenes KF707, introduction of a plasmid containing the RHA1 bphAaAb and KF707 bphA3A4 genes plus the RHA1 bphB2C1D1 genes into strain NK8. The remaining enzyme genes involved in the transformation of biphenyl to benzoate, bphB2C1D1, which encodes dehydrogenase, ring-cleavage dioxygenase and hydrolase, confer activities to NK8, installation of the polychlorinated biphenyls degradation pathway, overview
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in Escherichia coli strain M15 and SG13009; in Pseudomonas putida strain KT2442
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in Escherichia coli, strain BL21(DE3)
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in Escherichia coli, strain M15
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in Escherichia coli, strains M15 and SG13009
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in Pseudomonas putida strain KT2442
pET101[L11E10-bphAE] and pDB31[LB400-bphFGBC] cotransformed into Escherichia coli BL21 Star(DE3)
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pET101[LB400-bphAE] and pDB31[LB400-bphFGBC] cotransformed into Escherichia coli BL21 Star(DE3)
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the construction of appropriate hybrid genes may be used as a general strategy to overcome problems in obtaining heterologous biphenyl dioxygenase activities in Escherichia coli or other host organisms
the evolved bphA1 gene, in which nine amino acids from the Pseudomonas pseudoalcaligenes KF707 BphA1 are changed to those from the Burkholderia xenovorans LB400 BphA1 (M247I/H255Q/V258I/G268A/D303E/-313G/S324T/V325I/T376N), is expressed in Escherichia coli along with the bphA2A3A4 and bphB genes derived from strain KF707. This recombinant Escherichia coli cells convert biphenyl and several heterocyclic aromatic compounds into the highly hydroxylated products such as biphenyl-2,3,2',3'-tetraol (from biphenyl), 2-(2,3-dihydroxyphenyl)benzoxazole-4,5-diol (from 2-phenylbenzoxazole), and 2-(2,5-dihydroxyphenyl)benzoxazole-4,5-diol (from 2-(2-hydroxyphenyl)benzoxazole)
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tobacco plants transiently expressing BPDO
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EXPRESSION
ORGANISM
UNIPROT
LITERATURE
bph gene cluster bphAaAbAcAdCB is localized on the linear plasmid pRHL1, which is followed by bphS and bphT genes which encode a two-component signal transduction system composed of a BphT response regulator and a BpdS sensor kinase and promote transcriptional induction by aromatic compounds biphenyl, benzene, alkylbenzene and chlorinated benzene
exposure of soil microcosms to different polychlorinated biphenyl congeners results in different expression patterns of biphenyl dioxygenase genes
exposure to polychlorinated biphenyl and Aroclor 1242 results in a marked increase of two out of the four BPH genes. Exposure of soil microcosms to different polychlorinated biphenyl congeners results in different expression patterns of biphenyl dioxygenase genes
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ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
A234S
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significant change in regiospecificity of substrate dioxygenation, factor 2.3
F332A
-
significant change in regiospecificity of substrate dioxygenation
F378A
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very strong change in regiospecificity of substrate dioxygenation, factor higher than 7
F384A
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very strong change in regiospecificity of substrate dioxygenation, factor higher than 7
G389A
-
significant change in regiospecificity of substrate dioxygenation
I243A
-
significant change in regiospecificity of substrate dioxygenation
I326A
-
significant change in regiospecificity of substrate dioxygenation
M231A
-
very strong change in regiospecificity of substrate dioxygenation, factor higher than 7
M324A
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significant change in regiospecificity of substrate dioxygenation
N377A
-
significant change in regiospecificity of substrate dioxygenation
P334A
-
significant change in regiospecificity of substrate dioxygenation
T335G/F336I/N338T/I341T
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relaxation of the enzyme towards polychlorinated biphenyls. Wild-type enzyme shows less than 10% degradation with 2,6-dichlorobiphenyl, 3,3'-dichlorobiphenyl, 4,4'-dichlorobiphenyl, 2,3',4'-trichlorobiphenyl - the mutant enzyme enzyme shows 45 to 99% depletion depending on the substrate. Wild-type enzyme shows no degradation of 2,4,4'-trichlorobiphenyl, mutant enzyme shows 44% depletion
V383S
-
significant change in regiospecificity of substrate dioxygenation
V393S
-
significant change in regiospecificity of substrate dioxygenation
W342A
-
significant change in regiospecificity of substrate dioxygenation
W392A
-
significant change in regiospecificity of substrate dioxygenation
A234S
-
significant change in regiospecificity of substrate dioxygenation, factor 2.3
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I243A
-
significant change in regiospecificity of substrate dioxygenation
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I326A
-
significant change in regiospecificity of substrate dioxygenation
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M231A
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very strong change in regiospecificity of substrate dioxygenation, factor higher than 7
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N377A
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significant change in regiospecificity of substrate dioxygenation
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T335G/F336I/N338T/I341T
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relaxation of the enzyme towards polychlorinated biphenyls. Wild-type enzyme shows less than 10% degradation with 2,6-dichlorobiphenyl, 3,3'-dichlorobiphenyl, 4,4'-dichlorobiphenyl, 2,3',4'-trichlorobiphenyl - the mutant enzyme enzyme shows 45 to 99% depletion depending on the substrate. Wild-type enzyme shows no degradation of 2,4,4'-trichlorobiphenyl, mutant enzyme shows 44% depletion
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N348H
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wild-type enzyme shows less than 10% activity with 2,6-dichlorobiphenyl, 4,4'-dichlorobiphenyl - mutant enzyme shows about 55% depletion. Wild-type enzyme shows no degradation of 2,4,4'-trichlorobiphenyl and 2,2',5,5'-tetrachlorobiphenyl - mutant enzyme shows 50% and 92% depletion
N348H/A404V
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wild-type enzyme shows less than 10% activity with 2,6-dichlorobiphenyl, 4,4'-dichlorobiphenyl - mutant enzyme shows about 55% depletion. Wild-type enzyme shows no degradation of 2,4,4'-trichlorobiphenyl and 2,2',5,5'-tetrachlorobiphenyl - mutant enzyme shows 29% and 84% depletion
T375N
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conversion of sequence to corresponding sequence of Pseudomonas sp. strain LB400
F336M
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the mutant produces principally 3,4-dihydro-3,4-dihydroxy-2,2'-dichlorobiphenyl from 2,2'-dichlorobiphenyl
F370Y
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lower reactivity toward 2,2-dichlorobiphenyl but unchanged regiospecificity toward this substrate compared to the wild type enzyme
L283S
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lower reactivity toward 2,2-dichlorobiphenyl but unchanged regiospecificity toward this substrate compared to the wild type enzyme
M237T
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lower reactivity toward 2,2-dichlorobiphenyl but unchanged regiospecificity toward this substrate compared to the wild type enzyme
S238T
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lower reactivity toward 2,2-dichlorobiphenyl but unchanged regiospecificity toward this substrate compared to the wild type enzyme
T335A/F336I
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the ratio of the product formed from 2,2'-dichlorobiphenyl, 2,3-dihydroxy-2'-chlorobiphenyl to 3,4-dihydro-3,4-dihydroxy-2,2'-dichlorobiphenyl is: 90/10 for the wild-type enzyme and 40/60 for the mutant enzymes
T335A/F336L
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the ratio of the product formed from 2,2'-dichlorobiphenyl, 2,3-dihydroxy-2'-chlorobiphenyl to 3,4-dihydro-3,4-dihydroxy-2,2'-dichlorobiphenyl is: 90/10 for the wild-type enzyme and 85/15 for the mutant enzymes
T335A/F336L/I341V
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the ratio of the product formed from 2,2'-dichlorobiphenyl, 2,3-dihydroxy-2'-chlorobiphenyl to 3,4-dihydro-3,4-dihydroxy-2,2'-dichlorobiphenyl is: 90/10 for the wild-type enzyme and 40/60 for the mutant enzymes
T335A/F336M
T335G
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the ratio of the product formed from 2,2'-dichlorobiphenyl, 2,3-dihydroxy-2'-chlorobiphenyl to 3,4-dihydro-3,4-dihydroxy-2,2'-dichlorobiphenyl is: 90/10 for the wild-type enzyme and 80/20 for the mutant enzymes
T377N
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lower reactivity toward 2,2-dichlorobiphenyl but unchanged regiospecificity toward this substrate compared to the wild type enzyme
H255Q/V258I/G268A/F277Y
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mutant of the bphA1 gene encoding the large subunit, that is responsible for substrate specificity, extremely enhanced benzene-, toluene-, and alkylbenzene-degrading ability
I335F/T376N
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does not show any significant difference in the oxidation of biphenyl compared with wild type Bph Dox, and exhibits 2,3-dioxygenase activity for 2,2'-dichlorobiphenyl and 3,4-dioxygenase activity for 2,5,4'-trichlorobiphenyl
T376D
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decreased degradation activity
T376F
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shows novel degradation activity for dibenzofuran, which is a poor substrate for the wild type enzyme
T376G
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decreased degradation activity
T376K
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shows novel degradation activity for dibenzofuran, which is a poor substrate for the wild type enzyme
T376N
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shows novel degradation activity for dibenzofuran, which is a poor substrate for the wild type enzyme
T376S
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decreased degradation activity
T376V
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shows novel degradation activity for dibenzofuran, which is a poor substrate for the wild type enzyme
H255Q/V258I/G268A/F277Y
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mutant of the bphA1 gene encoding the large subunit, that is responsible for substrate specificity, extremely enhanced benzene-, toluene-, and alkylbenzene-degrading ability
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T376F
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shows novel degradation activity for dibenzofuran, which is a poor substrate for the wild type enzyme
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T376G
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decreased degradation activity
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T376K
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shows novel degradation activity for dibenzofuran, which is a poor substrate for the wild type enzyme
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T376N
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shows novel degradation activity for dibenzofuran, which is a poor substrate for the wild type enzyme
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T376V
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shows novel degradation activity for dibenzofuran, which is a poor substrate for the wild type enzyme
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T335A/F336T/N338T/I341T
additional information
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
synthesis
Show AA Sequence (210 entries)
Please use the Sequence Search for a specific query.