Any feedback?
Information on EC 1.14.13.208 - benzoyl-CoA 2,3-epoxidase and Organism(s) Aromatoleum evansii and UniProt Accession Q9AIX7
for references in articles please use BRENDA:EC1.14.13.208Please wait a moment until all data is loaded. This message will disappear when all data is loaded.
EC Tree
1.14.13.208 benzoyl-CoA 2,3-epoxidaseIUBMB Comments
The enzyme is involved in aerobic benzoate metabolism in Azoarcus evansii. BoxB functions as the oxygenase part of benzoyl-CoA oxygenase in conjunction with BoxA, the reductase component, which upon binding of benzoyl-CoA, transfers two electrons to the ring in the course of monooxygenation. BoxA is a homodimeric 46 kDa iron-sulfur-flavoprotein (FAD), BoxB is a monomeric iron-protein .
Specify your search results
Word Map
- 1.14.13.208
- evansii
- benzoate
-
azoarcus
-
epoxidation
-
diiron
- boxc
- thioesters
- protocatechuate
- enoyl-coa
-
homodimeric
-
non-aromatic
- oxygenases
-
coa-dependent
- succinyl-coa
-
dihydrodiol
-
iron-sulfur
- semialdehyde
- phenylacetyl-coa
-
benzoate-induced
-
diferric
-
benzoate-coa
The taxonomic range for the selected organisms is: Aromatoleum evansii
The expected taxonomic range for this enzyme is: Bacteria, Archaea
The expected taxonomic range for this enzyme is: Bacteria, Archaea
Synonyms
benzoyl-coa oxygenase, boxba, benzoyl-coa epoxidase, boxab, benzoyl-coa 2,3-epoxidase, benzoyl-coa dioxygenase/reductase, benzoyl coenzyme a epoxidase, 
more
topSYNONYM 
ORGANISM
UNIPROT
COMMENTARY 
LITERATURE
BoxB
BoxB
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT
LITERATURE
benzoyl-CoA + NADPH + H+ + O2 = 2,3-epoxy-2,3-dihydrobenzoyl-CoA + NADP+ + H2O
epoxidation starts with the binding of the O2 molecule to the diferrous center to generate a diferric peroxide complex, followed by concerted O-O bond cleavage and epoxide formation. Two different pathways have been located, leading to (2S,3R)-epoxy and (2R,3S)-epoxy products. The barrier difference is 2.8 kcal/mol, corresponding to a diastereomeric excess of about 99:1. Further isomerization from epoxide to phenol has quite a high barrier, which cannot compete with the product release step. After product release into solution, fast epoxide-oxepin isomerization and racemization can take place easily, leading to a racemic mixture of (2S,3R) and (2R,3S) products. The deoxygenation of epoxide to regenerate benzoyl-CoA by a diferrous form of the enzyme proceeds via a stepwise mechanism. The C2-O bond cleavage happens first, coupled with one electron transfer from one iron center to the substrate, to form a radical intermediate, which is followed by the second C3-O bond cleavage. The first step is rate-limiting. Reaction mechanism, modeling and simulations, detailed overview
benzoyl-CoA + NADPH + H+ + O2 = 2,3-epoxy-2,3-dihydrobenzoyl-CoA + NADP+ + H2O
molecular reaction mechanism analysis, different pathways are possible, detailed overview
-
SYSTEMATIC NAME
IUBMB Comments
benzoyl-CoA,NADPH:oxygen oxidoreductase (2,3-epoxydizing)
The enzyme is involved in aerobic benzoate metabolism in Azoarcus evansii. BoxB functions as the oxygenase part of benzoyl-CoA oxygenase in conjunction with BoxA, the reductase component, which upon binding of benzoyl-CoA, transfers two electrons to the ring in the course of monooxygenation. BoxA is a homodimeric 46 kDa iron-sulfur-flavoprotein (FAD), BoxB is a monomeric iron-protein [1].
SUBSTRATE
PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
benzoyl-CoA + NADPH + H+ + O2
2,3-epoxy-2,3-dihydrobenzoyl-CoA + NADP+ + H2O
-
-
-
?
2-fluorobenzoyl-CoA + NADPH + H+ + O2
?
83% of the activity with benzoyl-CoA
-
-
?
4-fluorobenzoyl-CoA + NADPH + H+ + O2
?
30% of the activity with benzoyl-CoA
-
-
?
benzoyl-CoA + NADPH + H+ + O2
2,3-epoxy-2,3-dihydrobenzoyl-CoA + NADP+ + H2O
-
-
-
-
?
benzoyl-CoA + NADPH + H+ + O2
?
-
second step in aerobic benzoate metabolism, enzyme is induced by benzoate
-
-
?
benzoyl-CoA + NADPH + H+ + O2
2,3-dihydro-2,3-dihydroxybenzoyl-CoA + NADP+
the enzyme is involved in aerobic benzoate metabolism in Azoarcus evansii
-
-
?
benzoyl-CoA + NADPH + H+ + O2
2,3-dihydro-2,3-dihydroxybenzoyl-CoA + NADP+
no activity with NADH as electron donor
-
-
?
additional information
?
-
benzoyl-CoA epoxidase is a dinuclear iron enzyme that catalyzes the epoxidation reaction of the aromatic ring of benzoyl-CoA with chemo-, regio- and stereo-selectivity. The enzyme may also catalyze the deoxygenation reaction of epoxide, suggesting a unique bifunctionality among the diiron enzymes
-
-
?
additional information
?
-
BoxB functions as the oxygenase part of benzoyl-CoA oxygenase in conjunction with BoxA, the reductase component. BoxA functions as the reducing component of benzoyl-CoA oxygenase, which, upon binding of benzoyl-CoA, transfers two electrons to the ring in the course of dioxygenation
-
-
?
additional information
?
-
-
BoxB functions as the oxygenase part of benzoyl-CoA oxygenase in conjunction with BoxA, the reductase component. BoxA functions as the reducing component of benzoyl-CoA oxygenase, which, upon binding of benzoyl-CoA, transfers two electrons to the ring in the course of dioxygenation
-
-
?
additional information
?
-
no activity with benzoate or phenylacetyl-CoA
-
-
?
additional information
?
-
-
no activity with benzoate or phenylacetyl-CoA
-
-
?
additional information
?
-
-
purified reductase component BoxA catalyses, in the absence of BoxB, the electron transfer from NADPH to free FAD with a rate of 17 mmol/min*mg protein. This activity is very high compared to the benzoyl-CoA conversion rate with purified BoxAB (0.29 mmol/min*mg BoxA). The reductase component BoxA oxidizes NADPH to transfer electrons, mediated by enzyme-bound FAD, to one or two of the [4Fe-4S] centres of the protein. The flow of electrons from NADPH to the artificial electron acceptor, free FAD, is greatly (10-fold) stimulated by binding of benzoyl-CoA to BoxA
-
-
?
additional information
?
-
-
benzoyl-CoA epoxidase is a dinuclear iron enzyme that catalyzes the epoxidation reaction of the aromatic ring of benzoyl-CoA with chemo-, regio- and stereo-selectivity
-
-
?
NATURAL SUBSTRATE
NATURAL PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
benzoyl-CoA + NADPH + H+ + O2
2,3-epoxy-2,3-dihydrobenzoyl-CoA + NADP+ + H2O
-
-
-
?
benzoyl-CoA + NADPH + H+ + O2
2,3-dihydro-2,3-dihydroxybenzoyl-CoA + NADP+
the enzyme is involved in aerobic benzoate metabolism in Azoarcus evansii
-
-
?
benzoyl-CoA + NADPH + H+ + O2
2,3-epoxy-2,3-dihydrobenzoyl-CoA + NADP+ + H2O
-
-
-
-
?
benzoyl-CoA + NADPH + H+ + O2
?
-
second step in aerobic benzoate metabolism, enzyme is induced by benzoate
-
-
?
additional information
?
-
benzoyl-CoA epoxidase is a dinuclear iron enzyme that catalyzes the epoxidation reaction of the aromatic ring of benzoyl-CoA with chemo-, regio- and stereo-selectivity. The enzyme may also catalyze the deoxygenation reaction of epoxide, suggesting a unique bifunctionality among the diiron enzymes
-
-
?
additional information
?
-
-
benzoyl-CoA epoxidase is a dinuclear iron enzyme that catalyzes the epoxidation reaction of the aromatic ring of benzoyl-CoA with chemo-, regio- and stereo-selectivity
-
-
?
COFACTOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
FAD
-
the reductase componment BoxA contains 0.72 mol flavin adenine dinucleotide. BoxA oxidizes NADPH to transfer electrons, mediated by enzyme-bound FAD, to one or two of the [4Fe-4S] centres of the protein. The flow of electrons from NADPH to the artificial electron acceptor, free FAD, is greatly (10-fold) stimulated by binding of benzoyl-CoA to BoxA
NADPH
-
the reductase component BoxA oxidizes NADPH to transfer electrons, mediated by enzyme-bound FAD, to one or two of the [4Fe-4S] centres of the protein. The flow of electrons from NADPH to the artificial electron acceptor, free FAD, is greatly (10-fold) stimulated by binding of benzoyl-CoA to BoxA. No oxidation of NADH
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
Fe2+
Fe
Fe2+
Fe2+
a non-heme diiron enzyme. The two iron ions are bridged by a glutamate (Glu150). The first Fe2+ is ligated by a glutamate (Glu120) and a histidine (His153), while the second Fe2+ is ligated by an aspartate (Asp211), a glutamate (Glu240) and a histidine (His243)
Fe
BoxA is a homodimeric 46 kDa iron-sulfur-flavoprotein, which acts as reductase. BoxB is a monomeric iron-protein. Subunit BoxB contains 1.6 mol of iron per mol of protein, addition of 1 mM Fe2+ to the assays has no stimulatory or inhibitory effect on the catalytic turnover
Fe
-
the reductase componment BoxA contains 10.4 to 18.4 mol of Fe, and 13.3 to 17.9 mol of acid-labile sulfur per mol of native protein, depending on the method of protein determination
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
KCl
higher salt concentration (500 mM KCl) results in sixfold lower activity suggesting that the interaction of the protein components is affected by high salt concentration
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
BoxC
addition of the putative ring cleaving enzyme BoxC leads to a several-fold acceleration of the initial rate and completes conversion of benzoyl-CoA. BoxC might facilitate the binding of BoxA to BoxB and thus lead to the observed rate increase
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.03
benzoyl-CoA
pH 8, 22°C, addition of the putative ring cleaving enzyme BoxC leads to a several-fold acceleration of the initial rate and completes conversion of benzoyl-CoA. Because of this complex behaviour of the oxygenase system, the apparent Km value for benzoyl-CoA can only be estimated from the time curve of benzoyl-CoA concentration in an assay mixture that contains BoxAB and the putative ring-cleaving enzyme BoxC. This curve shows a half-maximal rate at a benzoyl-CoA concentration of 0.03 mM
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
pI VALUE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
5.59
benzoyl-CoA oxygenase component A, calculated from sequence
5.6
5.62
benzoyl-CoA oxygenase component B, calculated from sequence
ORGANISM
COMMENTARY
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
SOURCE
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
additional information
additional information
benzoyl-CoA substrate forms two hydrogen bonds with Gln116, which in turn is hydrogen-bonded to Glu120. A number of other second-shell residues are also important for the orientation of the benzoyl moiety, including Thr119, Ser123, Phe193, Phe203, and Thr210. Optimized structure of the BoxB active site with the truncated benzoyl-CoA and O2 substrates bound, corresponding to the Michaelis complex, enzyme structure and reaction mechanism mechanics/molecular mechanics calculations and modeling, overview
additional information
-
usage of PDB ID 3PM5 for enzyme structure modeling and simulation, quantum mechanics/molecular mechanics calculations, overview. Four general pathways for oxidizing aromatic rings are determined
UNIPROT
ENTRY NAME
ORGANISM
NO. OF AA
NO. OF TRANSM. HELICES
MOLECULAR WEIGHT[Da]
SOURCE
SEQUENCE
LOCALIZATION PREDICTION
The reliability class of the localization prediction ranges from 1 (very reliable) to 5 (not reliable).
The reliability class of the localization prediction ranges from 1 (very reliable) to 5 (not reliable). MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
46000
46000
2 * 46000 (BoxA) + 50000 (BoxB), boxA/boxB system, BoxA is a homodimeric 46 kDa iron-sulfur-flavoprotein, which acts as reductase. BoxB is a monomeric iron-protein (50000-60000 Da, gel filtration. 50000 Da, SDS-PAGE)
46000
x + 55000 (benzoyl-CoA oxygenase component B) + x * 46000, (benzoyl-CoA oxygenase component A), BoxB functions as the oxygenase part of benzoyl-CoA oxygenase in conjunction with BoxA, the reductase component. BoxA functions as the reducing component of benzoyl-CoA oxygenase, which, upon binding of benzoyl-CoA, transfers two electrons to the ring in the course of dioxygenation, calculated from sequence
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
?
2 * 46000 (BoxA) + 50000 (BoxB), boxA/boxB system, BoxA is a homodimeric 46 kDa iron-sulfur-flavoprotein, which acts as reductase. BoxB is a monomeric iron-protein (50000-60000 Da, gel filtration. 50000 Da, SDS-PAGE)
?
x + 55000 (benzoyl-CoA oxygenase component B) + x * 46000, (benzoyl-CoA oxygenase component A), BoxB functions as the oxygenase part of benzoyl-CoA oxygenase in conjunction with BoxA, the reductase component. BoxA functions as the reducing component of benzoyl-CoA oxygenase, which, upon binding of benzoyl-CoA, transfers two electrons to the ring in the course of dioxygenation, calculated from sequence
CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
hanging drop vapor diffusion method, using 1.1 M NaH2PO4 and 0.2 M K2HPO4 as precipitant
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
His6-tagged component B of benzoyl-CoA oxygenase, component A of benzoyl-CoA oxygenase
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Mohamed, M.E.; Zaar, A.; Ebenau-Jehle, C.; Fuchs, G.
Reinvestigation of a new type of aerobic benzoate metabolism in the proteobacterium Azoarcus evansii
J. Bacteriol.
183
1899-1908
2001
Aromatoleum evansii
Gescher, J.; Zaar, A.; Mohamed, M.; Schgger, H.; Fuchs, G.
Genes coding for a new pathway of aerobic benzoate metabolism in Azoarcus evansii
J. Bacteriol.
184
6301-6315
2002
Aromatoleum evansii (Q9AIX6 and Q9AIX7), Aromatoleum evansii
Zaar, A.; Gescher, J.; Eisenreich, W.; Bacher, A.; Fuchs, G.
New enzymes involved in aerobic benzoate metabolism in Azoarcus evansii
Mol. Microbiol.
54
223-238
2004
Aromatoleum evansii (Q9AIX6 and Q9AIX7), Aromatoleum evansii, Aromatoleum evansii KB740 (Q9AIX6 and Q9AIX7)
Rather, L.J.; Knapp, B.; Haehnel, W.; Fuchs, G.
Coenzyme A-dependent aerobic metabolism of benzoate via epoxide formation
J. Biol. Chem.
285
20615-20624
2010
Aromatoleum evansii, Aromatoleum evansii KB740
Rather, L.J.; Bill, E.; Ismail, W.; Fuchs, G.
The reducing component BoxA of benzoyl-coenzyme A epoxidase from Azoarcus evansii is a [4Fe-4S] protein
Biochim. Biophys. Acta
1814
1609-1615
2011
Aromatoleum evansii
Rather, L.J.; Weinert, T.; Demmer, U.; Bill, E.; Ismail, W.; Fuchs, G.; Ermler, U.
Structure and mechanism of the diiron benzoyl-coenzyme A epoxidase BoxB
J. Biol. Chem.
286
29241-29248
2011
Aromatoleum evansii (Q9AIX7), Aromatoleum evansii
Liao, R.; Siegbahn, P.
Mechanism and selectivity of the dinuclear iron benzoyl-coenzyme A epoxidase BoxB
Chem. Sci.
6
2754-2764
2015
Aromatoleum evansii (Q9AIX7)
EXTERNAL LINKS (specific for EC-Number 1.14.13.208)
Use of this online version of BRENDA is free under the CC BY 4.0 license. See terms of use for full details.
Release 2023.1 (January 2023)
Legal
Curated at
Member of
