Information on EC 1.14.12.3 - benzene 1,2-dioxygenase:

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The expected taxonomic range for this enzyme is: Bacteria


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EC NUMBERCOMMENTARY
1.14.12.3-

RECOMMENDED NAMEGeneOntology No.
benzene 1,2-dioxygenaseGO:0018619

REACTIONREACTION DIAGRAMCOMMENTARYORGANISM UNIPROT ACCESSION NO.LITERATURE
benzene + NADH + H+ + O2 = cis-cyclohexa-3,5-diene-1,2-diol + NAD+
show the reaction diagram
----
benzene + NADH + H+ + O2 = cis-cyclohexa-3,5-diene-1,2-diol + NAD+
show the reaction diagram
the catalytic iron-sulfur proteins of the enzyme consist of two dissimilar alpha and beta subunits, the alpha subunit contains a [2Fe-2S] cluster involved in electron transfer, the catalytic nonheme iron center, and is also responsible for substrate specificityPseudomonas putida-657604

REACTION TYPEORGANISM UNIPROT ACCESSION NO.COMMENTARYLITERATURE
oxidation----
redox reaction----
reduction----

PATHWAYKEGG LinkMetaCyc Link
benzene degradation-PWY-5450
Benzoate degradation00362 -
Chlorocyclohexane and chlorobenzene degradation00361 -
Microbial metabolism in diverse environments01120 -

SYSTEMATIC NAMEIUBMB Comments
benzene,NADH:oxygen oxidoreductase (1,2-hydroxylating)A system, containing a reductase which is an iron-sulfur flavoprotein (FAD), an iron-sulfur oxygenase and ferredoxin. Requires Fe2+.

SYNONYMSORGANISM UNIPROT ACCESSION NO.COMMENTARYLITERATURE
BDO----
BDOPseudomonas putida--439072
BDOPseudomonas putida ML2---
BEDPseudomonas putida--711842
BEDPseudomonas putida MST---
benzene dioxygenase----
benzene dioxygenasePseudomonas putida--439072, 657604, 711842
benzene dioxygenasePseudomonas putida ML2, Pseudomonas putida MST---
benzene hydroxylase----
oxygenase, benzene 1,2-di-----
benzene hydroxylaseRhodococcus opacusQ58818-674962
additional informationPseudomonas putida-enzyme system consisting of three components: a flavoprotein reductase, a ferredoxin and a catalytic iron sulfur protein657604

CAS REGISTRY NUMBERCOMMENTARY
9075-66-5-

ORGANISMCOMMENTARYLITERATURESEQUENCE CODESEQUENCE DB SOURCE
Pseudomonas putida-439063, 439065, 439066, 657604, 711842--Manually annotated by BRENDA team
Pseudomonas putidastrain ML2439064, 439068, 439069, 439070, 439071--Manually annotated by BRENDA team
Pseudomonas putidastrain ML2; strain ML2, genes bedC1C2BA439072--Manually annotated by BRENDA team
Pseudomonas putida ML2strain ML2439064, 439068, 439069, 439070, 439071--Manually annotated by BRENDA team
Pseudomonas putida ML2strain ML2; strain ML2, genes bedC1C2BA439072--Manually annotated by BRENDA team
Pseudomonas putida MST-711842--Manually annotated by BRENDA team
Pseudomonas sp.-439067--Manually annotated by BRENDA team
Rhodococcus opacuslarge subunit; strains B-4, B-9 and B-10674962Q58819TrEMBLManually annotated by BRENDA team
Rhodococcus opacussmall subunit; strains B-4, B-9 and B-10674962Q58818TrEMBLManually annotated by BRENDA team

GENERAL INFORMATIONORGANISM UNIPROT ACCESSION NO.COMMENTARYLITERATURE
No entries in this field

SUBSTRATEPRODUCT                      REACTION DIAGRAMORGANISM UNIPROT ACCESSION NO. COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
1-methylcyclohexene + O2 + NADH?
show the reaction diagram
Pseudomonas putida, Pseudomonas putida ML2--439072--?
2 cyclohexene + 2 O2 + 2 NADH + 2 H+2-cyclohexen-1-ol + cis-1,2-cyclohexanediol + 2 NAD+ + H2O
show the reaction diagram
Pseudomonas putida--439072failure of transformed bacteria to grow on cyclohexene is attributed to the toxicity of metabolic intermediates accumulating from the 2-cyclohexen-1-ol metabolism, cyclohexene metabolism pathways, overview-?
3-methylcyclohexene + O2 + NADH?
show the reaction diagram
Pseudomonas putida, Pseudomonas putida ML2--439072--?
benzene + NADH + H+ + O2cis-cyclohexa-3,5-diene-1,2-diol + NAD+
show the reaction diagram
Pseudomonas putida, Pseudomonas putida MST--711842--?
benzene + NADH + O2cis-cyclohexa-3,5-diene-1,2-diol + NAD+
show the reaction diagram
Pseudomonas sp.--439067-439067?
benzene + NADH + O2cis-cyclohexa-3,5-diene-1,2-diol + NAD+
show the reaction diagram
Pseudomonas putida--439063-439063?
benzene + NADH + O2cis-cyclohexa-3,5-diene-1,2-diol + NAD+
show the reaction diagram
Pseudomonas putida--439064-439064?
benzene + NADH + O2cis-cyclohexa-3,5-diene-1,2-diol + NAD+
show the reaction diagram
Pseudomonas putida--439065-439065?
benzene + NADH + O2cis-cyclohexa-3,5-diene-1,2-diol + NAD+
show the reaction diagram
Pseudomonas putida--439066-439066?
benzene + NADH + O2cis-cyclohexa-3,5-diene-1,2-diol + NAD+
show the reaction diagram
Pseudomonas putida--439068-439068?
benzene + NADH + O2cis-cyclohexa-3,5-diene-1,2-diol + NAD+
show the reaction diagram
Pseudomonas putida--439069-439069?
benzene + NADH + O2cis-cyclohexa-3,5-diene-1,2-diol + NAD+
show the reaction diagram
Pseudomonas putida--439070-439070?
benzene + NADH + O2cis-cyclohexa-3,5-diene-1,2-diol + NAD+
show the reaction diagram
Pseudomonas putida--439071-439071?
benzene + NADH + O2cis-cyclohexa-3,5-diene-1,2-diol + NAD+
show the reaction diagram
Rhodococcus opacusQ58818, Q58819-674962--?
benzene + NADH + O2cis-cyclohexa-3,5-diene-1,2-diol + NAD+
show the reaction diagram
Pseudomonas putida--439072the enzyme oxidizes cyclohexene to a mixture of two products, a monohydroxylated 2-cyclohexen-1-ol product and a dihydroxylated cis-1,2-cyclohexanediol439072?
benzene + NADH + O2cis-cyclohexa-3,5-diene-1,2-diol + NAD+
show the reaction diagram
Pseudomonas putida ML2--439064-439064?
benzene + NADH + O2cis-cyclohexa-3,5-diene-1,2-diol + NAD+
show the reaction diagram
Pseudomonas putida ML2--439068-439068?
benzene + NADH + O2cis-cyclohexa-3,5-diene-1,2-diol + NAD+
show the reaction diagram
Pseudomonas putida ML2--439069-439069?
benzene + NADH + O2cis-cyclohexa-3,5-diene-1,2-diol + NAD+
show the reaction diagram
Pseudomonas putida ML2--439070-439070?
benzene + NADH + O2cis-cyclohexa-3,5-diene-1,2-diol + NAD+
show the reaction diagram
Pseudomonas putida ML2--439071-439071?
benzene + NADH + O2cis-cyclohexa-3,5-diene-1,2-diol + NAD+
show the reaction diagram
Pseudomonas putida ML2--439072the enzyme oxidizes cyclohexene to a mixture of two products, a monohydroxylated 2-cyclohexen-1-ol product and a dihydroxylated cis-1,2-cyclohexanediol439072?
benzene + NADH + O2?
show the reaction diagram
Pseudomonas sp.-reaction in benzene catabolism439067--?
benzene + NADH + O2?
show the reaction diagram
Pseudomonas putida-reaction in benzene catabolism439063, 439064, 439065, 439066, 439068, 439069, 439070--?
benzene + NADH + O2?
show the reaction diagram
Pseudomonas putida ML2-reaction in benzene catabolism439064, 439068, 439069, 439070--?
benzene + NADH + O2(1R,2S)-cis-cyclohexa-3,5-diene-1,2-diol + NAD+
show the reaction diagram
Pseudomonas putida-bioremediation of aromatic environmental pollutants, initial step of benzene degradation657604--?
benzene + NADPH + O2cis-cyclohexa-3,5-diene-1,2-diol + NAD+
show the reaction diagram
Pseudomonas sp.-activity with NADPH is 10% of the activity with NADH439067--?
benzene + O2 + NADHcis-cyclohexa-3,5-diene-1,2-diol + NAD+
show the reaction diagram
Pseudomonas putida--439072, 657604--?
benzene + O2 + NADHcis-cyclohexa-3,5-diene-1,2-diol + NAD+
show the reaction diagram
Pseudomonas putida-best substrate439072--?
benzene + O2 + NADHcis-cyclohexa-3,5-diene-1,2-diol + NAD+
show the reaction diagram
Pseudomonas putida ML2-best substrate439072--?
ethylbenzene + NADH + O2cis-2,3-dihydroxy-1-ethyl-cyclohexa-4,6-diene + 1-phenethyl alcohol + NAD+
show the reaction diagram
Pseudomonas putida--657604--?
ethylbenzene + O2 + NADHcis-2,3-dihydroxy-1-ethyl-cyclohexa-4,6-diene + NAD+
show the reaction diagram
Pseudomonas putida--657604--?
toluene + NADH + O2cis-2,3-dihydroxy-1-methyl-cyclohexa-4,6-diene + NAD+
show the reaction diagram
Pseudomonas putida--657604--?
toluene + O2 + NADH?
show the reaction diagram
Pseudomonas putida--439072--?
trifluoromethylbenzene + NADH + O2?
show the reaction diagram
Pseudomonas putida, Pseudomonas putida ML2--439069---

NATURAL SUBSTRATESNATURAL PRODUCTSREACTION DIAGRAMORGANISM UNIPROT ACCESSION NO.COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
1-methylcyclohexene + O2 + NADH?
show the reaction diagram
Pseudomonas putida, Pseudomonas putida ML2--439072--?
2 cyclohexene + 2 O2 + 2 NADH + 2 H+2-cyclohexen-1-ol + cis-1,2-cyclohexanediol + 2 NAD+ + H2O
show the reaction diagram
Pseudomonas putida--439072failure of transformed bacteria to grow on cyclohexene is attributed to the toxicity of metabolic intermediates accumulating from the 2-cyclohexen-1-ol metabolism, cyclohexene metabolism pathways, overview-?
3-methylcyclohexene + O2 + NADH?
show the reaction diagram
Pseudomonas putida, Pseudomonas putida ML2--439072--?
benzene + NADH + H+ + O2cis-cyclohexa-3,5-diene-1,2-diol + NAD+
show the reaction diagram
Pseudomonas putida, Pseudomonas putida MST--711842--?
benzene + NADH + O2?
show the reaction diagram
Pseudomonas sp.-reaction in benzene catabolism439067--?
benzene + NADH + O2?
show the reaction diagram
Pseudomonas putida-reaction in benzene catabolism439063, 439064, 439065, 439066, 439068, 439069, 439070--?
benzene + NADH + O2(1R,2S)-cis-cyclohexa-3,5-diene-1,2-diol + NAD+
show the reaction diagram
Pseudomonas putida-bioremediation of aromatic environmental pollutants, initial step of benzene degradation657604--?
benzene + NADH + O2?
show the reaction diagram
Pseudomonas putida ML2-reaction in benzene catabolism439064, 439068, 439069, 439070--?
benzene + O2 + NADHcis-cyclohexa-3,5-diene-1,2-diol + NAD+
show the reaction diagram
Pseudomonas putida, Pseudomonas putida ML2--439072--?
toluene + O2 + NADH?
show the reaction diagram
Pseudomonas putida--439072--?

COFACTORORGANISM UNIPROT ACCESSION NO.COMMENTARYLITERATUREIMAGE
FADPseudomonas putida-contains FAD439069 2D-image
FerredoxinPseudomonas putida-the benzene dioxygenase comprises three components: 1. flavoprotein, NADH:ferredoxin oxidoreductase, 2. an intermediate electron-transfer protein, or ferredoxin and 3. a terminal dioxygenase439066, 439069, 439070-
FerredoxinPseudomonas putida--657604-
iron-sulfur centrePseudomonas putida-[2Fe-2S] cluster657604 2D-image
NADHPseudomonas sp.--439067 2D-image
NADHRhodococcus opacusQ58818, Q58819-674962 2D-image
NADHPseudomonas putida--439063, 439064, 439065, 439066, 439068, 439069, 439072, 657604, 711842 2D-image
NADPHPseudomonas sp.-activity with NADPH is 10% of the activity with NADH439067 2D-image

METALS and IONS ORGANISM UNIPROT ACCESSION NO.COMMENTARY LITERATURE
Fe2+Pseudomonas putida-the benzene dioxygenase comprises three components: 1. flavoprotein, NADH:ferredoxin oxidoreductase, 2. an intermediate electron-transfer protein, or ferredoxin with a [2Fe-2S] cluster and 3. a terminal dioxygenase, containing two [2Fe-2S]iron-sulfur clusters which require two additional Fe2+ atoms/molecules for oxygenase activity439066
Fe2+Pseudomonas sp.-required439067
Fe2+Pseudomonas putida-the terminal dioxygenase component contains two Fe2+ ions per molecule in addition to two [2Fe-2S] iron-sulfur clusters439069
Fe2+Pseudomonas putida-required439072
Fe2+Pseudomonas putida-the alpha subunit of the enzyme contains a catalytic [2Fe-2S] cluster involved in electron transfer, the catalytic nonheme iron center is also responsible for substrate specificity, overview657604
IronPseudomonas putida-the intermediate electron-carrying protein possesses on [2Fe-2S] cluster, the terminal dioxygenase possesses 2 [2Fe-2S] clusters439063
IronPseudomonas putida-the terminal dioxygenase protein [2Fe-2S] centres, in the oxidized form the two iron atoms within the centre are high-spin ferric439065
IronPseudomonas putida-the benzene dioxygenase comprises three components: 1. flavoprotein, NADH:ferredoxin oxidoreductase, 2. an intermediate electron-transfer protein, or ferredoxin with a [2Fe-2S] cluster and 3. a terminal dioxygenase, containing two [2Fe-2S]iron-sulfur clusters which require two additional Fe2+ atoms/molecules for oxygenase activity439066
IronPseudomonas sp.-the terminal dioxygenase component contains 2 atoms of iron and 3 atoms of inorganic sulfur439067
IronPseudomonas putida-the sequence of the ferredoxin component of the benzene dioxygenase contains five Cys residues, four of which are required to coordinate the iron-sulfur cluster439068
IronPseudomonas putida-the terminal dioxygenase component contains two Fe2+ ions per molecule in addition to two [2Fe-2S] iron-sulfur clusters. The ferredoxin component contains one [2Fe-2S] cluster439069
IronPseudomonas putida-Rieske-type [2Fe-2S] centres are coordinated by two histidines and two cysteines439071

INHIBITORSORGANISM UNIPROT ACCESSION NO. COMMENTARY LITERATURE IMAGE
No entries in this field

ACTIVATING COMPOUNDORGANISM UNIPROT ACCESSION NO. COMMENTARY LITERATURE IMAGE
No entries in this field

KM VALUE [mM]KM VALUE [mM] MaximumSUBSTRATEORGANISM UNIPROT ACCESSION NO. COMMENTARY LITERATURE IMAGE
0.00782-NADHPseudomonas putida--439069 2D-image
0.0112-NADHPseudomonas sp.--439067 2D-image

TURNOVER NUMBER [1/s] TURNOVER NUMBER MAXIMUM[1/s] SUBSTRATEORGANISM UNIPROT ACCESSION NO. COMMENTARY LITERATURE IMAGE
No entries in this field

kcat/KM VALUE [1/mMs-1]kcat/KM VALUE [1/mMs-1] MaximumSUBSTRATEORGANISM UNIPROT ACCESSION NO. COMMENTARY LITERATURE IMAGE
No entries in this field

Ki VALUE [mM]Ki VALUE [mM] MaximumINHIBITORORGANISM UNIPROT ACCESSION NO. COMMENTARY LITERATURE IMAGE
No entries in this field

IC50 VALUE [mM]IC50 VALUE [mM] MaximumINHIBITORORGANISM UNIPROT ACCESSION NO. COMMENTARY LITERATURE IMAGE
No entries in this field

SPECIFIC ACTIVITY [µmol/min/mg] SPECIFIC ACTIVITY MAXIMUM ORGANISM UNIPROT ACCESSION NO. COMMENTARY LITERATURE
0.0049-Pseudomonas putida-recombinant enzyme expressed in Escherichia coli, substrate cyclohexene439072
0.0077-Pseudomonas putida-recombinant enzyme expressed in Escherichia coli, substrate 1-methylcyclohexene439072
0.0086-Pseudomonas putida-recombinant enzyme expressed in Escherichia coli, substrate 3-methylcyclohexene439072
0.0172-Pseudomonas putida-recombinant enzyme expressed in Escherichia coli, substrate toluene439072
0.0236-Pseudomonas putida-recombinant enzyme expressed in Escherichia coli, substrate benzene439072
0.025-Pseudomonas putida-with ethylbenzene as substrate657604
0.071-Pseudomonas putida-with toluene as substrate657604
0.115-Pseudomonas putida-with benzene as substrate657604
additional information-Pseudomonas sp.--439067

pH OPTIMUMpH MAXIMUMORGANISM UNIPROT ACCESSION NO. COMMENTARYLITERATURE
7-Pseudomonas putida-assay at711842
7.2-Pseudomonas putida-assay at439072, 657604

pH RANGEpH RANGE MAXIMUMORGANISM UNIPROT ACCESSION NO.COMMENTARYLITERATURE
No entries in this field

TEMPERATURE OPTIMUMTEMPERATURE OPTIMUM MAXIMUMORGANISM UNIPROT ACCESSION NO.COMMENTARYLITERATURE
22-Pseudomonas putida-assay at room temperature657604
30-Pseudomonas putida-assay at439072, 711842

TEMPERATURE RANGE TEMPERATURE MAXIMUM ORGANISM UNIPROT ACCESSION NO. COMMENTARY LITERATURE
No entries in this field

pI VALUEpI VALUE MAXIMUMORGANISM UNIPROT ACCESSION NO.COMMENTARYLITERATURE
No entries in this field

SOURCE TISSUE ORGANISM UNIPROT ACCESSION NO. COMMENTARY LITERATURE SOURCE
No entries in this field

LOCALIZATION ORGANISM UNIPROT ACCESSION NO. COMMENTARY GeneOntology No. LITERATURE SOURCE
solublePseudomonas sp.---439067Manually annotated by BRENDA team

PDBSCOPCATHORGANISM
No entries in this field

MOLECULAR WEIGHT MOLECULAR WEIGHT MAXIMUM ORGANISM UNIPROT ACCESSION NO. COMMENTARY LITERATURE
11860-Pseudomonas putida-ferredoxin component of the benzene dioxygenase, fast atom bombardment mass spectrometry439068
12000-Pseudomonas putida-intermediate electron-carrying protein, gel filtration439063
12300-Pseudomonas putida-intermediate electron-carrying protein, meniscus depletion method439063
168000-Pseudomonas putida-gel filtration439064
186000-Pseudomonas sp.-terminal dioxygenase component, meniscus depletion method439067
215000-Pseudomonas putida-terminal dioxygenase component, meniscus depletion method439066
215300-Pseudomonas putida-terminal dioxygenase component, meniscus depletion method439063

SUBUNITS ORGANISM UNIPROT ACCESSION NO. COMMENTARY LITERATURE
?Pseudomonas putida-alpha2, beta2, 2 * 54500 + 2 * 23500, terminal dioxygenase component. 2 * 42000, reductase component. The ferredoxin component has a MW of 12300 Da439069
?Pseudomonas putida ML2-alpha2, beta2, 2 * 54500 + 2 * 23500, terminal dioxygenase component. 2 * 42000, reductase component. The ferredoxin component has a MW of 12300 Da-
tetramerPseudomonas putida-2 * 23500 + 2 * 54500, SDS-PAGE439064
tetramerPseudomonas putida ML2-2 * 23500 + 2 * 54500, SDS-PAGE-
dimerPseudomonas putida-alpha, beta, iron sulfur protein component; the enzyme consist of two dissimilar alpha and beta subunits, alpha subunit structure modeling657604
additional informationPseudomonas putida-the benzene dioxygenase comprises three components: 1. flavoprotein, NADH:ferredoxin oxidoreductase, 2. an intermediate electron-transfer protein, or ferredoxin and 3. a terminal dioxygenase439066, 439069
additional informationPseudomonas putida-55000 Da is the MW of the alpha-subunit of the terminal dioxygenase439070
additional informationPseudomonas putida ML2-55000 Da is the MW of the alpha-subunit of the terminal dioxygenase; the benzene dioxygenase comprises three components: 1. flavoprotein, NADH:ferredoxin oxidoreductase, 2. an intermediate electron-transfer protein, or ferredoxin and 3. a terminal dioxygenase-

POSTTRANSLATIONAL MODIFICATION ORGANISM UNIPROT ACCESSION NO. COMMENTARY LITERATURE
No entries in this field

Crystallization/COMMENTARY ORGANISM UNIPROT ACCESSION NO. LITERATURE
No entries in this field

pH STABILITYpH STABILITY MAXIMUM ORGANISM UNIPROT ACCESSION NO. COMMENTARY LITERATURE
No entries in this field

TEMPERATURE STABILITYTEMPERATURE STABILITY MAXIMUM ORGANISM UNIPROT ACCESSION NO. COMMENTARYLITERATURE
No entries in this field

GENERAL STABILITYORGANISM UNIPROT ACCESSION NO.LITERATURE
No entries in this field

ORGANIC SOLVENT ORGANISM UNIPROT ACCESSION NO. COMMENTARY LITERATURE
No entries in this field

OXIDATION STABILITY ORGANISM UNIPROT ACCESSION NO. LITERATURE
No entries in this field

STORAGE STABILITY ORGANISM UNIPROT ACCESSION NO. LITERATURE
-20°C, stable for 6 months to 1 yearPseudomonas putida-439069

Purification/COMMENTARY ORGANISM UNIPROT ACCESSION NO. LITERATURE
-Pseudomonas putida-439069
iron-sulfur proteins of the benzene dioxygenase system: 1.intermediate electron-carrying protein and terminal dioxygenasePseudomonas putida-439063
-Pseudomonas sp.-439067

Cloned/COMMENTARY ORGANISM UNIPROT ACCESSION NO. LITERATURE
expressed in Escherichia coli JM109; expression of wild-type and mutant enzymes in Escherichia coli strains JM109 and CJ236Pseudomonas putida-657604
expression in Escherichia coli JM109; genes bedC1C2BA, under the control of the tac promoter, subcloned into pLAFR5, successfully conjugated into seven of the Gram-negative cis-1,2-cyclohexanediol-degrading isolates and stably maintained and expressed in three of them, strain CHD1–CHD3, the strains grow on cis-1,2-cyclohexanediol as sole carbon source, express an active BDO and oxidise cyclohexene, but none of the three strains is able to grow on cyclohexene as sole carbon source, overview, expression in Escherichia coli strain JM109Pseudomonas putida-439072
expression in Escherichia coli strain DH5alpha and JM109, co-expression with benzene dihydrodiol dehydrogenase under the control of the Ptac promoter or without any induction. The recombinant strains expressing the BED and the BDDH enzymes transform benzene into dihydrodiol with corresponding consumption of oxygen and regenerate NADH by converting dihydrodiol to catecholPseudomonas putida-711842
expression of the alpha-subunit and the beta-subunit of terminal dioxygenase in Escherichia coliPseudomonas putida-439071
genes bedC1 and bedC2 encoding the terminal oxygenase alpha-subunit and beta-subunit, expression in Escherichia coliPseudomonas putida-439070

EXPRESSION ORGANISM UNIPROT ACCESSION NO. LITERATURE
No entries in this field

ENGINEERINGORGANISM UNIPROT ACCESSION NO.COMMENTARYLITERATURE
A291SPseudomonas putida-reduced activity with ethylbenzene657604
E444DPseudomonas putida-no effect on activity657604
G404DPseudomonas putida-reduced activity with ethylbenzene657604
H119CPseudomonas putida-the mutant alpha-subunit of the terminal dioxygenase is unable to coordinate an EPR-detectable Rieske [2Fe-2S] cluster with the characteristic g factors439070
H222MPseudomonas putida-in the reconstitution assay with the reductase component, the ferredoxin component and the beta-subunit of terminal dioxygenase the mutant alpha-subunit is unable to reconstitute dioxygenase activity439070
H228CPseudomonas putida-in the reconstitution assay with the reductase component, the ferredoxin component and the beta-subunit of terminal dioxygenase the mutant alpha-subunit is unable to reconstitute dioxygenase activity439070
H98CPseudomonas putida-the mutant alpha-subunit of the terminal dioxygenase is unable to coordinate an EPR-detectable Rieske [2Fe-2S] cluster with the characteristic g factors, detection of a novel EPR spectrum, the intensity of the spectrum is approximately 8% from the wild-type439070
I301VPseudomonas putida-the mutation in the C-terminal part of subunit alpha enhances the substrate specificity for ethylbenzene, the mutant shows altered patterns of products formed from toluene and ethylbenzene, including monohydroxylated side chains657604
I301V/T305S/I307L/L309VPseudomonas putida-increased activity with ethylbenzene; the mutations in the C-terminal part of subunit alpha enhance the substrate specificity for ethylbenzene, the quadruple mutant also shows a high uncoupled rate of electron transfer without product formation657604
I307LPseudomonas putida-the mutation in the C-terminal part of subunit alpha enhances the substrate specificity for ethylbenzene, the mutant shows altered patterns of products formed from toluene and ethylbenzene, including monohydroxylated side chains657604
I412VPseudomonas putida-reduced activity with ethylbenzene657604
K436RPseudomonas putida-reduced activity with ethylbenzene657604
L285WPseudomonas putida-reduced activity with ethylbenzene657604
L285W/A291S/G404DPseudomonas putida-slightly reduced activity with ethylbenzene657604
L28W/A291SPseudomonas putida-reduced activity with ethylbenzene657604
T305SPseudomonas putida-the mutation in the C-terminal part of subunit alpha enhances the substrate specificity for ethylbenzene, the mutant shows altered patterns of products formed from toluene and ethylbenzene, including monohydroxylated side chains657604
V324I/I327VPseudomonas putida-reduced activity with ethylbenzene657604
Y118SPseudomonas putida-the mutant alpha-subunit of the terminal dioxygenase shows an EPR spectrum of half the intensity of that of the wild-type. In the reconstitution assay with the reductase component, the ferredoxin component and the beta-subunit of terminal dioxygenase it shows significantly reduced activities439070
Y221APseudomonas putida-in the reconstitution assay with the reductase component, the ferredoxin component and the beta-subunit of terminal dioxygenase the mutant alpha-subunit shows significantly reduced activity439070
H119CPseudomonas putida ML2-the mutant alpha-subunit of the terminal dioxygenase is unable to coordinate an EPR-detectable Rieske [2Fe-2S] cluster with the characteristic g factors-
H222MPseudomonas putida ML2-in the reconstitution assay with the reductase component, the ferredoxin component and the beta-subunit of terminal dioxygenase the mutant alpha-subunit is unable to reconstitute dioxygenase activity-
H98CPseudomonas putida ML2-the mutant alpha-subunit of the terminal dioxygenase is unable to coordinate an EPR-detectable Rieske [2Fe-2S] cluster with the characteristic g factors, detection of a novel EPR spectrum, the intensity of the spectrum is approximately 8% from the wild-type-
Y118SPseudomonas putida ML2-the mutant alpha-subunit of the terminal dioxygenase shows an EPR spectrum of half the intensity of that of the wild-type. In the reconstitution assay with the reductase component, the ferredoxin component and the beta-subunit of terminal dioxygenase it shows significantly reduced activities-
L309VPseudomonas putida-the mutation in the C-terminal part of subunit alpha enhances the substrate specificity for ethylbenzene, the mutant shows altered patterns of products formed from toluene and ethylbenzene, including monohydroxylated side chains657604
additional informationPseudomonas putida-construction of chimeric proteins and mutants of the benzene dioxygenase alpha subunit, the chimera are formed by benzene and toluene dioxygenases, the amino acid sequences of the alpha subunits of both enzymes differ at only 33 of 450 amino acids, these residues are primarily responsible for the change in specificity, the chimeric protein containing toluene dioxygenase C-terminal region residues 281 to 363 shows greater substrate preference for alkyl benzenes, identification of four amino acid substitutions in this region, I301V, T305S, I307L, and L309V, that particularly enhance the preference for ethylbenzene, structure modeling, overview657604
additional informationPseudomonas putida-establishing of a detection system to monitor environmental benzene contamination by co-expression of benzene dioxygenase with benzene dihydrodiol dehydrogenase in Escherichia coli. The procedures involving whole-cell bioassays determine the concentration of benzene through benzene dioxygenase activity, which allows for direct correlation of oxygen consumption, and through the benzene dihydrodiol dehydrogenase that causes catechol accumulation and restores NADH necessary for the activity of the first enzyme711842
Y221APseudomonas putida ML2-in the reconstitution assay with the reductase component, the ferredoxin component and the beta-subunit of terminal dioxygenase the mutant alpha-subunit shows significantly reduced activity-
additional informationPseudomonas putida MST-establishing of a detection system to monitor environmental benzene contamination by co-expression of benzene dioxygenase with benzene dihydrodiol dehydrogenase in Escherichia coli. The procedures involving whole-cell bioassays determine the concentration of benzene through benzene dioxygenase activity, which allows for direct correlation of oxygen consumption, and through the benzene dihydrodiol dehydrogenase that causes catechol accumulation and restores NADH necessary for the activity of the first enzyme-

Renatured/COMMENTARYORGANISM UNIPROT ACCESSION NO.LITERATURE
No entries in this field

APPLICATIONORGANISM UNIPROT ACCESSION NO.COMMENTARYLITERATURE
analysisPseudomonas putida-the enzyme is used in a whole-cell assay as biosensor for benzene concentration and vapopur, overview711842
analysisPseudomonas putida MST-the enzyme is used in a whole-cell assay as biosensor for benzene concentration and vapopur, overview-

REF. AUTHORS TITLE JOURNAL VOL. PAGES YEAR ORGANISM (UNIPROT ACCESSION NO.)LINK TO PUBMEDSOURCE
439063Crutcher, S.E.; Geary, P.J.Properties of the iron--sulphur proteins of the benzene dioxygenase system from Pseudomonas putidaBiochem. J.177393-4001979Pseudomonas putida PubMed
439064Zamanian, M.; Mason, J.R.Benzene dioxygenase in Pseudomonas putida. Subunit composition and immuno-cross-reactivity with other aromatic dioxygenasesBiochem. J.244611-6161987Pseudomonas putida, Pseudomonas putida ML2 PubMed
439065Geary, P.J.; Dickson, D.P.Moessbauer spectroscopic studies of the terminal dioxygenase protein of benzene dioxygenase from Pseudomonas putidaBiochem. J.195199-2031981Pseudomonas putida PubMed
439066Geary, P.J.; Saboowalla, F.; Patil, D.; Cammack, R.An investigation of the iron-sulphur proteins of benzene dioxygenase from Pseudomonas putida by electron-spin-resonance spectroscopyBiochem. J.217667-6731984Pseudomonas putida PubMed
439067Axcell, B.C.; Geary, P.J.Purification and some properties of a soluble benzene-oxidizing system from a strain of PseudomonasBiochem. J.146173-1831975Pseudomonas sp. PubMed
439068Morrice, N.; Geary, P.J.; Cammack, R.; Harris, A.; Beg, F.; Aitken, A.Primary structure of protein B from Pseudomonas putida, member of a new class of 2Fe-2S ferredoxinsFEBS Lett.231336-3401988Pseudomonas putida, Pseudomonas putida ML2 PubMed
439069Wackett, L.P.Benzene dioxygenase from Pseudomonas putida, ML2 (NCIB 12190)Methods Enzymol.18852-601990Pseudomonas putida, Pseudomonas putida ML2 PubMed
439070Mason, J.R.; Butler, C.S.; Cammack, R.; Shergill, J.K.Structural studies on the catalytic component of benzene dioxygenase from Pseudomonas putidaBiochem. Soc. Trans.2590-951997Pseudomonas putida, Pseudomonas putida ML2 PubMed
439071Shergill, J.K.; Butler, C.S.; White, A.C.; Cammack, R.; Mason, J.R.EPR, ENDOR and ESEEM studies on recombinant benzene dioxygenaseBiochem. Soc. Trans.22288S1994Pseudomonas putida, Pseudomonas putida ML2 PubMed
439072Swift, R.J.; Carter, S.F.; Widdowson, D.A.; Mason, J.R.; Leak, D.J.Expression of benzene dioxygenase from Pseudomonas putida ML2 in cis-1,2-cyclohexanediol-degrading pseudomonadsAppl. Microbiol. Biotechnol.55721-7262001Pseudomonas putida, Pseudomonas putida ML2 PubMed
657604Bagneris, C.; Cammack, R.; Mason, J.R.Subtle difference between benzene and toluene dioxygenases of Pseudomonas putidaAppl. Environ. Microbiol.711570-15802005Pseudomonas putida PubMed
674962Na, K.S.; Kuroda, A.; Takiguchi, N.; Ikeda, T.; Ohtake, H.; Kato, J.Isolation and characterization of benzene-tolerant Rhodococcus opacus strainsJ. Biosci. Bioeng.99378-3822005Rhodococcus opacus (Q58818), Rhodococcus opacus (Q58819), Rhodococcus opacus PubMed
711842Di Gennaro, P.; Bruzzese, N.; Anderlini, D.; Aiossa, M.; Papacchini, M.; Campanella, L.; Bestetti, G.Development of microbial engineered whole-cell systems for environmental benzene determinationEcotoxicol. Environ. Saf.74542-5492010Pseudomonas putida, Pseudomonas putida MST PubMed

LINKS TO OTHER DATABASES (specific for EC-Number 1.14.12.3)
ExplorEnz
ExPASy
KEGG
MetaCyc
NCBI: PubMed, Protein, Nucleotide, Structure, Genome, OMIM
IUBMB Enzyme Nomenclature
PROSITE Database of protein families and domains
SYSTERS
Protein Mutant Database
InterPro (database of protein families, domains and functional sites)