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Information on EC 3.3.2.10 - soluble epoxide hydrolase and Organism(s) Aspergillus niger and UniProt Accession Q9UR30

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EC Tree
     3 Hydrolases
         3.3 Acting on ether bonds
             3.3.2 Ether hydrolases
                3.3.2.10 soluble epoxide hydrolase
IUBMB Comments
Catalyses the hydrolysis of trans-substituted epoxides, such as trans-stilbene oxide, as well as various aliphatic epoxides derived from fatty-acid metabolism . It is involved in the metabolism of arachidonic epoxides (epoxyicosatrienoic acids; EETs) and linoleic acid epoxides. The EETs, which are endogenous chemical mediators, act at the vascular, renal and cardiac levels to regulate blood pressure [4,5]. The enzyme from mammals is a bifunctional enzyme: the C-terminal domain exhibits epoxide-hydrolase activity and the N-terminal domain has the activity of EC 3.1.3.76, lipid-phosphate phosphatase [1,2]. Like EC 3.3.2.9, microsomal epoxide hydrolase, it is probable that the reaction involves the formation of an hydroxyalkyl---enzyme intermediate [4,6]. The enzyme can also use leukotriene A4, the substrate of EC 3.3.2.6, leukotriene-A4 hydrolase, but it forms 5,6-dihydroxy-7,9,11,14-icosatetraenoic acid rather than leukotriene B4 as the product [9,10]. In vertebrates, five epoxide-hydrolase enzymes have been identified to date: EC 3.3.2.6 (leukotriene-A4 hydrolase), EC 3.3.2.7 (hepoxilin-epoxide hydrolase), EC 3.3.2.9 (microsomal epoxide hydrolase), EC 3.3.2.10 (soluble epoxide hydrolase) and EC 3.3.2.11 (cholesterol 5,6-oxide hydrolase) .
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Aspergillus niger
UNIPROT: Q9UR30
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The taxonomic range for the selected organisms is: Aspergillus niger
The expected taxonomic range for this enzyme is: Eukaryota, Bacteria, Archaea
Reaction Schemes
hide
an epoxide
+
H2O
=
a glycol
+
=
Synonyms
epoxide hydrolase, soluble epoxide hydrolase, ephx2, cytosolic epoxide hydrolase, epoxide hydrolase 1, epoxide hydrolase 2, ephx3, teso hydrolase, hepoxilin hydrolase, bnseh1, more
SYNONYM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
an epoxide + H2O = a glycol
show the reaction diagram
the catalytic triad is formed by Asp192, Asp348, and His374
SYSTEMATIC NAME
IUBMB Comments
epoxide hydrolase
Catalyses the hydrolysis of trans-substituted epoxides, such as trans-stilbene oxide, as well as various aliphatic epoxides derived from fatty-acid metabolism [7]. It is involved in the metabolism of arachidonic epoxides (epoxyicosatrienoic acids; EETs) and linoleic acid epoxides. The EETs, which are endogenous chemical mediators, act at the vascular, renal and cardiac levels to regulate blood pressure [4,5]. The enzyme from mammals is a bifunctional enzyme: the C-terminal domain exhibits epoxide-hydrolase activity and the N-terminal domain has the activity of EC 3.1.3.76, lipid-phosphate phosphatase [1,2]. Like EC 3.3.2.9, microsomal epoxide hydrolase, it is probable that the reaction involves the formation of an hydroxyalkyl---enzyme intermediate [4,6]. The enzyme can also use leukotriene A4, the substrate of EC 3.3.2.6, leukotriene-A4 hydrolase, but it forms 5,6-dihydroxy-7,9,11,14-icosatetraenoic acid rather than leukotriene B4 as the product [9,10]. In vertebrates, five epoxide-hydrolase enzymes have been identified to date: EC 3.3.2.6 (leukotriene-A4 hydrolase), EC 3.3.2.7 (hepoxilin-epoxide hydrolase), EC 3.3.2.9 (microsomal epoxide hydrolase), EC 3.3.2.10 (soluble epoxide hydrolase) and EC 3.3.2.11 (cholesterol 5,6-oxide hydrolase) [7].
CAS REGISTRY NUMBER
COMMENTARY hide
9048-63-9
-
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
(R)-4-nitrostyrene oxide + H2O
?
show the reaction diagram
-
-
-
?
(R)-1-chloro-2-(2,4-difluorophenyl)-2,3-epoxypropane + H2O
(R)-3-chloro-2-(2,4-difluoro-phenyl)-propane-1,2-diol
show the reaction diagram
-
in a racemic mixture only the (R)-enantiomeric epoxide is converted to the (R)-enantiomeric diol
-
-
?
(R)-p-nitrostyrene oxide + H2O
1-(4-nitrophenyl)ethane-1,2-diol
show the reaction diagram
-
-
-
-
?
(R,S)-4-nitrostyrene oxide + H2O
1-(4-nitrophenyl)ethane-1,2-diol
show the reaction diagram
-
-
-
-
?
(S)-4-nitrostyrene oxide
1-(4-nitrophenyl)ethane-1,2-diol
show the reaction diagram
-
-
-
-
?
glycidyl phenyl ether + H2O
?
show the reaction diagram
-
the selectivity factor E reflecting the relative rate of the reaction of the two enantiomers is only 4.6 for the wild-type enzyme, in slight favor of the (S)-product, a value of 10.8 is observed with the natural EH variant IS002B1 with the three amino acid exchanges A217V, K332E and A390E. A value of 7.4 is observed with the natural variant IR003B1 with the amino acid exchange R219G, a value of 6.6 is observed with the natural variant IB001C2 with the amino acid exchange F340Y, a value of 6.4 is observed with the natural variant IE001H6 with the amino acid exchange A327V, a value of 6.2 is observed with the natural variant IS001H8 with the amino acid exchange A327V and a value of 5.5 is observed with the natural variant IL001D4 with the amino acid exchange P222S
-
-
?
p-nitrostyrene oxide + H2O
?
show the reaction diagram
-
-
-
-
?
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
4-hydroxymercuribenzoate
-
-
cis-limonene epoxide
-
weak
dithionitrobenzene
-
-
epichlorhydrin
-
weak
geraniol derivatives
-
weak
-
iodoacetamide
-
-
m-chloroperbenzoic acid
-
-
omega-bromo-nitro-acetophenone
-
-
trans-limonene epoxide
-
weak
trichloropropene oxide
-
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.6
(R)-p-nitrostyrene oxide
-
-
1
(R,S)-4-nitrostyrene oxide
-
-
-
3.7
(S)-4-nitrostyrene oxide
-
-
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
16.4
(R)-p-nitrostyrene oxide
-
-
13.9
(R,S)-4-nitrostyrene oxide
-
-
-
1.7
(S)-4-nitrostyrene oxide
-
-
-
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5.5 - 8
-
pH 5.5: about 35% of maximal activity, pH 8.0: about 45% of maximal activity
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
15 - 45
-
15°C: about 50% of maximal activity, 45°C: about 65% of maximal activity
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
the enzyme has no N-terminal membrane anchor in contrast to the microsomal epoxide hydrolase, 3.3.2.9
-
Manually annotated by BRENDA team
UNIPROT
ENTRY NAME
ORGANISM
NO. OF AA
NO. OF TRANSM. HELICES
MOLECULAR WEIGHT[Da]
SOURCE
SEQUENCE
LOCALIZATION PREDICTION?
Q9UR30_ASPNG
398
0
44524
TrEMBL
-
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
100000
recombinant enzyme, gel filtration
185000
-
gel filtration
45000
-
4 * 45000, SDS-PAGE
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
tetramer
-
4 * 45000, SDS-PAGE
additional information
the enzyme has no N-terminal membrane anchor in contrast to the microsomal epoxide hydrolase, EC 3.3.2.9
CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
three-dimensional crystal structure is determined at 3.5 A resolution by the multiwavelength anomalous diffraction method using crystals of a seleno-methionine substituted form of enzyme and then refined at 1.8 A resolution
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
D192A
site directed mutagenesis, inactive or nearly inactive mutant
D192N
site directed mutagenesis, inactive or nearly inactive mutant
D192S
site directed mutagenesis, inactive or nearly inactive mutant
D348A
site directed mutagenesis, inactive or nearly inactive mutant
D348E
site directed mutagenesis, the mutant shows 48% of wild-type enzyme activity
H374K
site directed mutagenesis, inactive or nearly inactive mutant
H374N
site directed mutagenesis, inactive or nearly inactive mutant
H374S
site directed mutagenesis, inactive or nearly inactive mutant, recombinantly expressed mutant enzyme is not soluble but remains in the particulate fraction of Escherichia coli cells
additional information
-
the selectivity factor E reflecting the relative rate of the reaction of the two enantiomers is only 4.6 for the wild-type enzyme, in slight favor of the (S)-product, a value of 10.8 is observed with the natural EH variant IS002B1 with the three amino acid exchanges A217V, K332E and A390E. A value of 7.4 is observed with the natural variant IR003B1 with the amino acid exchange R219G, a value of 6.6 is observed with the natural variant IB001C2 with the amino acid exchange F340Y, a value of 6.4 is observed with the natural variant IE001H6 with the amino acid exchange A327V, a value of 6.2 is observed with the natural variant IS001H8 with the amino acid exchange A327V and a value of 5.5 is observed with the natural variant IL001D4 with the amino acid exchange P222S
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
recombinant enzyme from Escherichia coli strain BL21(DE3) to homogeneity by ammonium sulfate fractionation, anion exchange and hydrophobic interaction chromatography, and gel filtration
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
DNA and amino acid sequence determination and analysis, phylogenetic tree, sequence comparisons, functional expression of wild-type enzyme and expression of enzyme mutants in Escherichia coli strain BL21(DE3)
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
synthesis
the enzyme is useful for enantioselective bio-organic synthesis of chiral substances
synthesis
-
the enzyme may be a good biocatalyst for the preparation of enantiopure epoxides or diols
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Morisseau, C.; Archelas, A.; Guitton, C.; Faucher, D.; Furstoss, R.; Baratti, J.C.
Purification and characterization of a highly enantioselective epoxide hydrolase from Aspergillus niger
Eur. J. Biochem.
263
386-395
1999
Aspergillus niger
Manually annotated by BRENDA team
Reetz, M.T.; Torre, C.; Eipper, A.; Lohmer, R.; Hermes, M.; Brunner, B.; Maichele, A.; Bocola, M.; Arand, M.; Cronin, A.; Genzel, Y.; Archelas, A.; Furstoss, R.
Enhancing the enantioselectivity of an epoxide hydrolase by directed evolution
Org. Lett.
6
177-180
2004
Aspergillus niger
Manually annotated by BRENDA team
Zou, J.; Hallberg, B.M.; Bergfors, T.; Oesch, F.; Arand, M.; Mowbray, S.L.; Jones, T.A.
Structure of Aspergillus niger epoxide hydrolase at 1.8 A resolution: implications for the structure and function of the mammalian microsomal class of epoxide hydrolases
Structure Fold. Des.
8
111-122
2000
Aspergillus niger (Q9UR30), Aspergillus niger, Aspergillus niger LCP521 (Q9UR30)
Manually annotated by BRENDA team
Monfort, N.; Archelas, A.; Furstoss, R.
Enzymatic transformations. Part 55: Highly productive epoxide hydrolase catalyzed resolution of an azole antifungal key synthon
Tetrahedron
60
601-605
2004
Aspergillus niger
-
Manually annotated by BRENDA team
Arand, M.; Hemmer, H.; Duerk, H.; Barattis, J.; Archelas, A.; Furstoss, R.
Cloning and molecular characterization of a soluble epoxide hydrolase from Aspergillus niger that is related to mammalian microsomal epoxide hydrolase
Biochem. J.
344
273-280
1999
Aspergillus niger (Q9UR30), Aspergillus niger LCP521 (Q9UR30)
-
Manually annotated by BRENDA team