Information on EC 1.14.12.3 -

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

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EC NUMBER	COMMENTARY
1.14.12.3	-

RECOMMENDED NAME	GeneOntology No.
benzene 1,2-dioxygenase	GO:0018619

REACTION	REACTION DIAGRAM	COMMENTARY	ORGANISM	UNIPROT ACCESSION NO.	LITERATURE
benzene + NADH + H+ + O2 = cis-cyclohexa-3,5-diene-1,2-diol + NAD+		-	-	-	-
benzene + NADH + H+ + O2 = cis-cyclohexa-3,5-diene-1,2-diol + NAD+		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 specificity	Pseudomonas putida	-	657604

REACTION TYPE	ORGANISM	UNIPROT ACCESSION NO.	COMMENTARY	LITERATURE
oxidation	-	-	-	-
redox reaction	-	-	-	-
reduction	-	-	-	-

PATHWAY	KEGG Link	MetaCyc Link
benzene degradation	-	PWY-5450

SYSTEMATIC NAME	IUBMB 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+.

SYNONYMS	ORGANISM	UNIPROT ACCESSION NO.	COMMENTARY	LITERATURE
BDO	-	-	-	-
BDO	Pseudomonas putida	-	-	439072
BED	Pseudomonas putida	-	-	711842
benzene dioxygenase	-	-	-	-
benzene dioxygenase	Pseudomonas putida	-	-	439072, 657604, 711842
benzene hydroxylase	-	-	-	-
oxygenase, benzene 1,2-di-	-	-	-	-
benzene hydroxylase	Rhodococcus opacus	Q58818, Q58819	-	674962
additional information	Pseudomonas putida	-	enzyme system consisting of three components: a flavoprotein reductase, a ferredoxin and a catalytic iron sulfur protein	657604

CAS REGISTRY NUMBER	COMMENTARY
9075-66-5	-

ORGANISM	COMMENTARY	LITERATURE	SEQUENCE CODE	SEQUENCE DB	SOURCE
Pseudomonas putida	-	439063, 439065, 439066, 657604, 711842	-	-
Pseudomonas putida	strain ML2	439064, 439068, 439069, 439070, 439071	-	-
Pseudomonas putida	strain ML2; strain ML2, genes bedC1C2BA	439072	-	-
Pseudomonas putida MST	-	711842	-	-
Pseudomonas sp.	-	439067	-	-
Rhodococcus opacus	large subunit; strains B-4, B-9 and B-10	674962	Q58819	TrEMBL
Rhodococcus opacus	small subunit; strains B-4, B-9 and B-10	674962	Q58818	TrEMBL

GENERAL INFORMATION	ORGANISM	UNIPROT ACCESSION NO.	COMMENTARY	LITERATURE
No entries in this field

SUBSTRATE	PRODUCT	REACTION DIAGRAM	ORGANISM	UNIPROT ACCESSION NO.	COMMENTARY/ Substrate	LITERATURE/ Substrate	COMMENTARY/ Product	LITERATURE/ Product	Reversibility r=reversible ir=irreversible ?=not specified
1-methylcyclohexene + O2 + NADH	?		Pseudomonas putida	-	-	439072	-	-	?
3-methylcyclohexene + O2 + NADH	?		Pseudomonas putida	-	-	439072	-	-	?
benzene + NADH + H+ + O2	cis-cyclohexa-3,5-diene-1,2-diol + NAD+		Pseudomonas putida	-	-	711842	-	-	?
benzene + NADH + O2	cis-cyclohexa-3,5-diene-1,2-diol + NAD+		Pseudomonas sp.	-	-	439067	-	439067	?
benzene + NADH + O2	cis-cyclohexa-3,5-diene-1,2-diol + NAD+		Pseudomonas putida	-	-	439063	-	439063	?
benzene + NADH + O2	cis-cyclohexa-3,5-diene-1,2-diol + NAD+		Pseudomonas putida	-	-	439064	-	439064	?
benzene + NADH + O2	cis-cyclohexa-3,5-diene-1,2-diol + NAD+		Pseudomonas putida	-	-	439065	-	439065	?
benzene + NADH + O2	cis-cyclohexa-3,5-diene-1,2-diol + NAD+		Pseudomonas putida	-	-	439066	-	439066	?
benzene + NADH + O2	cis-cyclohexa-3,5-diene-1,2-diol + NAD+		Pseudomonas putida	-	-	439068	-	439068	?
benzene + NADH + O2	cis-cyclohexa-3,5-diene-1,2-diol + NAD+		Pseudomonas putida	-	-	439069	-	439069	?
benzene + NADH + O2	cis-cyclohexa-3,5-diene-1,2-diol + NAD+		Pseudomonas putida	-	-	439070	-	439070	?
benzene + NADH + O2	cis-cyclohexa-3,5-diene-1,2-diol + NAD+		Pseudomonas putida	-	-	439071	-	439071	?
benzene + NADH + O2	cis-cyclohexa-3,5-diene-1,2-diol + NAD+		Rhodococcus opacus	Q58818, Q58819	-	674962	-	-	?
benzene + NADH + O2	cis-cyclohexa-3,5-diene-1,2-diol + NAD+		Pseudomonas putida	-	-	439072	the enzyme oxidizes cyclohexene to a mixture of two products, a monohydroxylated 2-cyclohexen-1-ol product and a dihydroxylated cis-1,2-cyclohexanediol	439072	?
benzene + NADH + O2	?		Pseudomonas sp.	-	reaction in benzene catabolism	439067	-	-	?
benzene + NADH + O2	?		Pseudomonas putida	-	reaction in benzene catabolism	439063, 439064, 439065, 439066, 439068, 439069, 439070	-	-	?
benzene + NADH + O2	(1R,2S)-cis-cyclohexa-3,5-diene-1,2-diol + NAD+		Pseudomonas putida	-	bioremediation of aromatic environmental pollutants, initial step of benzene degradation	657604	-	-	?
benzene + NADPH + O2	cis-cyclohexa-3,5-diene-1,2-diol + NAD+		Pseudomonas sp.	-	activity with NADPH is 10% of the activity with NADH	439067	-	-	?
benzene + O2 + NADH	cis-cyclohexa-3,5-diene-1,2-diol + NAD+		Pseudomonas putida	-	-	439072, 657604	-	-	?
benzene + O2 + NADH	cis-cyclohexa-3,5-diene-1,2-diol + NAD+		Pseudomonas putida	-	best substrate	439072	-	-	?
cyclohexene + O2 + NADH	2-cyclohexen-1-ol + cis-1,2-cyclohexanediol		Pseudomonas putida	-	-	439072	failure 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	-	?
ethylbenzene + NADH + O2	cis-2,3-dihydroxy-1-ethyl-cyclohexa-4,6-diene + 1-phenethyl alcohol + NAD+		Pseudomonas putida	-	-	657604	-	-	?
ethylbenzene + O2 + NADH	cis-2,3-dihydroxy-1-ethyl-cyclohexa-4,6-diene + NAD+		Pseudomonas putida	-	-	657604	-	-	?
toluene + NADH + O2	cis-2,3-dihydroxy-1-methyl-cyclohexa-4,6-diene + NAD+		Pseudomonas putida	-	-	657604	-	-	?
toluene + O2 + NADH	?		Pseudomonas putida	-	-	439072	-	-	?
trifluoromethylbenzene + NADH + O2	?		Pseudomonas putida	-	-	439069	-	-	-

NATURAL SUBSTRATES	NATURAL PRODUCTS	REACTION DIAGRAM	ORGANISM	UNIPROT ACCESSION NO.	COMMENTARY SUBSTRATE	LITERATURE (Substrate)	COMMENTARY PRODUCT	LITERATURE (Product)
1-methylcyclohexene + O2 + NADH	?		Pseudomonas putida	-	-	439072	-	-
3-methylcyclohexene + O2 + NADH	?		Pseudomonas putida	-	-	439072	-	-
benzene + NADH + H+ + O2	cis-cyclohexa-3,5-diene-1,2-diol + NAD+		Pseudomonas putida	-	-	711842	-	-
benzene + NADH + O2	?		Pseudomonas sp.	-	reaction in benzene catabolism	439067	-	-
benzene + NADH + O2	?		Pseudomonas putida	-	reaction in benzene catabolism	439063, 439064, 439065, 439066, 439068, 439069, 439070	-	-
benzene + NADH + O2	(1R,2S)-cis-cyclohexa-3,5-diene-1,2-diol + NAD+		Pseudomonas putida	-	bioremediation of aromatic environmental pollutants, initial step of benzene degradation	657604	-	-
benzene + O2 + NADH	cis-cyclohexa-3,5-diene-1,2-diol + NAD+		Pseudomonas putida	-	-	439072	-	-
cyclohexene + O2 + NADH	2-cyclohexen-1-ol + cis-1,2-cyclohexanediol		Pseudomonas putida	-	-	439072	failure 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	-
toluene + O2 + NADH	?		Pseudomonas putida	-	-	439072	-	-

COFACTOR	ORGANISM	UNIPROT ACCESSION NO.	COMMENTARY	LITERATURE	IMAGE
FAD	Pseudomonas putida	-	contains FAD	439069	2D-image
Ferredoxin	Pseudomonas putida	-	the benzene dioxygenase comprises three components: 1. flavoprotein, NADH:ferredoxin oxidoreductase, 2. an intermediate electron-transfer protein, or ferredoxin and 3. a terminal dioxygenase	439066, 439069, 439070	-
Ferredoxin	Pseudomonas putida	-	-	657604	-
iron-sulfur centre	Pseudomonas putida	-	[2Fe-2S] cluster	657604	-
NADH	Pseudomonas putida	-	-	439063, 439064, 439065, 439066, 439068, 439069, 439072, 657604, 711842	2D-image
NADH	Pseudomonas sp.	-	-	439067	2D-image
NADH	Rhodococcus opacus	Q58818, Q58819	-	674962	2D-image
NADPH	Pseudomonas sp.	-	activity with NADPH is 10% of the activity with NADH	439067	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 activity	439066
Fe2+	Pseudomonas sp.	-	required	439067
Fe2+	Pseudomonas putida	-	the terminal dioxygenase component contains two Fe2+ ions per molecule in addition to two [2Fe-2S] iron-sulfur clusters	439069
Fe2+	Pseudomonas putida	-	required	439072
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, overview	657604
Iron	Pseudomonas putida	-	the intermediate electron-carrying protein possesses on [2Fe-2S] cluster, the terminal dioxygenase possesses 2 [2Fe-2S] clusters	439063
Iron	Pseudomonas putida	-	the terminal dioxygenase protein [2Fe-2S] centres, in the oxidized form the two iron atoms within the centre are high-spin ferric	439065
Iron	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 activity	439066
Iron	Pseudomonas sp.	-	the terminal dioxygenase component contains 2 atoms of iron and 3 atoms of inorganic sulfur	439067
Iron	Pseudomonas 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 cluster	439068
Iron	Pseudomonas 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] cluster	439069
Iron	Pseudomonas putida	-	Rieske-type [2Fe-2S] centres are coordinated by two histidines and two cysteines	439071

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

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

KM VALUE [mM]	KM VALUE [mM] Maximum	SUBSTRATE	ORGANISM	UNIPROT ACCESSION NO.	COMMENTARY	LITERATURE	IMAGE
0.00782	-	NADH	Pseudomonas putida	-	-	439069	2D-image
0.0112	-	NADH	Pseudomonas sp.	-	-	439067	2D-image

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

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

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

IC50 VALUE [mM]	IC50 VALUE [mM] Maximum	INHIBITOR	ORGANISM	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 cyclohexene	439072
0.0077	-	Pseudomonas putida	-	recombinant enzyme expressed in Escherichia coli, substrate 1-methylcyclohexene	439072
0.0086	-	Pseudomonas putida	-	recombinant enzyme expressed in Escherichia coli, substrate 3-methylcyclohexene	439072
0.0172	-	Pseudomonas putida	-	recombinant enzyme expressed in Escherichia coli, substrate toluene	439072
0.0236	-	Pseudomonas putida	-	recombinant enzyme expressed in Escherichia coli, substrate benzene	439072
0.025	-	Pseudomonas putida	-	with ethylbenzene as substrate	657604
0.071	-	Pseudomonas putida	-	with toluene as substrate	657604
0.115	-	Pseudomonas putida	-	with benzene as substrate	657604
additional information	-	Pseudomonas sp.	-	-	439067

pH OPTIMUM	pH MAXIMUM	ORGANISM	UNIPROT ACCESSION NO.	COMMENTARY	LITERATURE
7	-	Pseudomonas putida	-	assay at	711842
7.2	-	Pseudomonas putida	-	assay at	439072, 657604

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

TEMPERATURE OPTIMUM	TEMPERATURE OPTIMUM MAXIMUM	ORGANISM	UNIPROT ACCESSION NO.	COMMENTARY	LITERATURE
22	-	Pseudomonas putida	-	assay at room temperature	657604
30	-	Pseudomonas putida	-	assay at	439072, 711842

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

pI VALUE	pI VALUE MAXIMUM	ORGANISM	UNIPROT ACCESSION NO.	COMMENTARY	LITERATURE
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
soluble	Pseudomonas sp.	-	-	-	439067

PDB	SCOP	CATH	ORGANISM
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 spectrometry	439068
12000	-	Pseudomonas putida	-	intermediate electron-carrying protein, gel filtration	439063
12300	-	Pseudomonas putida	-	intermediate electron-carrying protein, meniscus depletion method	439063
168000	-	Pseudomonas putida	-	gel filtration	439064
186000	-	Pseudomonas sp.	-	terminal dioxygenase component, meniscus depletion method	439067
215000	-	Pseudomonas putida	-	terminal dioxygenase component, meniscus depletion method	439066
215300	-	Pseudomonas putida	-	terminal dioxygenase component, meniscus depletion method	439063

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 Da	439069
tetramer	Pseudomonas putida	-	2 * 23500 + 2 * 54500, SDS-PAGE	439064
dimer	Pseudomonas putida	-	alpha, beta, iron sulfur protein component; the enzyme consist of two dissimilar alpha and beta subunits, alpha subunit structure modeling	657604
additional information	Pseudomonas putida	-	the benzene dioxygenase comprises three components: 1. flavoprotein, NADH:ferredoxin oxidoreductase, 2. an intermediate electron-transfer protein, or ferredoxin and 3. a terminal dioxygenase	439066, 439069
additional information	Pseudomonas putida	-	55000 Da is the MW of the alpha-subunit of the terminal dioxygenase	439070

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

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

GENERAL STABILITY	ORGANISM	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 year	Pseudomonas 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 dioxygenase	Pseudomonas 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 CJ236	Pseudomonas 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 JM109	Pseudomonas 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 catechol	Pseudomonas putida	-	711842
expression of the alpha-subunit and the beta-subunit of terminal dioxygenase in Escherichia coli	Pseudomonas putida	-	439071
genes bedC1 and bedC2 encoding the terminal oxygenase alpha-subunit and beta-subunit, expression in Escherichia coli	Pseudomonas putida	-	439070

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

ENGINEERING	ORGANISM	UNIPROT ACCESSION NO.	COMMENTARY	LITERATURE
A291S	Pseudomonas putida	-	reduced activity with ethylbenzene	657604
E444D	Pseudomonas putida	-	no effect on activity	657604
G404D	Pseudomonas putida	-	reduced activity with ethylbenzene	657604
H119C	Pseudomonas 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	439070
H222M	Pseudomonas 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 activity	439070
H228C	Pseudomonas 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 activity	439070
H98C	Pseudomonas 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-type	439070
I301V	Pseudomonas 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 chains	657604
I301V/T305S/I307L/L309V	Pseudomonas 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 formation	657604
I307L	Pseudomonas 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 chains	657604
I412V	Pseudomonas putida	-	reduced activity with ethylbenzene	657604
K436R	Pseudomonas putida	-	reduced activity with ethylbenzene	657604
L285W	Pseudomonas putida	-	reduced activity with ethylbenzene	657604
L285W/A291S/G404D	Pseudomonas putida	-	slightly reduced activity with ethylbenzene	657604
L28W/A291S	Pseudomonas putida	-	reduced activity with ethylbenzene	657604
T305S	Pseudomonas 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 chains	657604
V324I/I327V	Pseudomonas putida	-	reduced activity with ethylbenzene	657604
Y118S	Pseudomonas 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 activities	439070
Y221A	Pseudomonas 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 activity	439070
L309V	Pseudomonas 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 chains	657604
additional information	Pseudomonas 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, overview	657604
additional information	Pseudomonas 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 enzyme	711842

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

APPLICATION	ORGANISM	UNIPROT ACCESSION NO.	COMMENTARY	LITERATURE
analysis	Pseudomonas putida	-	the enzyme is used in a whole-cell assay as biosensor for benzene concentration and vapopur, overview	711842

REF.	AUTHORS	TITLE	JOURNAL	VOL.	PAGES	YEAR	ORGANISM	LINK TO PUBMED	SOURCE
439063	Crutcher, S.E.; Geary, P.J.	Properties of the iron--sulphur proteins of the benzene dioxygenase system from Pseudomonas putida	Biochem. J.	177	393-400	1979	Pseudomonas putida	PubMed
439064	Zamanian, M.; Mason, J.R.	Benzene dioxygenase in Pseudomonas putida. Subunit composition and immuno-cross-reactivity with other aromatic dioxygenases	Biochem. J.	244	611-616	1987	Pseudomonas putida	PubMed
439065	Geary, P.J.; Dickson, D.P.	Moessbauer spectroscopic studies of the terminal dioxygenase protein of benzene dioxygenase from Pseudomonas putida	Biochem. J.	195	199-203	1981	Pseudomonas putida	PubMed
439066	Geary, 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 spectroscopy	Biochem. J.	217	667-673	1984	Pseudomonas putida	PubMed
439067	Axcell, B.C.; Geary, P.J.	Purification and some properties of a soluble benzene-oxidizing system from a strain of Pseudomonas	Biochem. J.	146	173-183	1975	Pseudomonas sp.	PubMed
439068	Morrice, 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 ferredoxins	FEBS Lett.	231	336-340	1988	Pseudomonas putida	PubMed
439069	Wackett, L.P.	Benzene dioxygenase from Pseudomonas putida, ML2 (NCIB 12190)	Methods Enzymol.	188	52-60	1990	Pseudomonas putida	PubMed
439070	Mason, J.R.; Butler, C.S.; Cammack, R.; Shergill, J.K.	Structural studies on the catalytic component of benzene dioxygenase from Pseudomonas putida	Biochem. Soc. Trans.	25	90-95	1997	Pseudomonas putida	PubMed
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439072	Swift, 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 pseudomonads	Appl. Microbiol. Biotechnol.	55	721-726	2001	Pseudomonas putida	PubMed
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711842	Di Gennaro, P.; Bruzzese, N.; Anderlini, D.; Aiossa, M.; Papacchini, M.; Campanella, L.; Bestetti, G.	Development of microbial engineered whole-cell systems for environmental benzene determination	Ecotoxicol. Environ. Saf.	74	542-549	2010	Pseudomonas putida	PubMed

LINKS TO OTHER DATABASES (specific for EC-Number 1.14.12.3)

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)

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