Information on EC 1.14.14.149 - 5-epiaristolochene 1,3-dihydroxylase

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The enzyme appears in viruses and cellular organisms

EC NUMBER
COMMENTARY hide
1.14.14.149
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RECOMMENDED NAME
GeneOntology No.
5-epiaristolochene 1,3-dihydroxylase
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REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
5-epiaristolochene + 2 [reduced NADPH-hemoprotein reductase] + 2 O2 = capsidiol + 2 [oxidized NADPH-hemoprotein reductase] + 2 H2O
show the reaction diagram
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REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
hydroxylation
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PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
Sesquiterpenoid and triterpenoid biosynthesis
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SYSTEMATIC NAME
IUBMB Comments
5-epiaristolochene,[reduced NADPH-hemoprotein reductase]:oxygen oxidoreductase (1- and 3-hydroxylating)
A heme-thiolate protein (P-450). Kinetic studies suggest that 1beta-hydroxyepiaristolochene is mainly formed first followed by hydroxylation at C-3. However the reverse order via 3alpha-hydroxyepiaristolochene does occur.
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
cvs. CM334 and ECW30R
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Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
evolution
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the enzyme is encoded by gene EAH which belongs to the multigene family EAS/EAH
metabolism
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the enzyme is involved in biosynthesis of capsidiol. Capsidiol accumulation is essential for nonhost resistance of pepper against Phytophthora infestans infection
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
1-deoxycapsidiol + [reduced NADPH-hemoprotein reductase] + O2
capsidiol + [oxidized NADPH-hemoprotein reductase] + H2O
show the reaction diagram
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-
-
-
?
1beta-hydroxy-5-epiaristolochene + [reduced NADPH-hemoprotein reductase] + O2
capsidiol + [oxidized NADPH-hemoprotein reductase] + H2O
show the reaction diagram
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the catalytic efficiency for 1beta-hydroxy-5-epiaristolochene is about 10times greater than that for 3alpha-hydroxy-5-epiaristolochene
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?
3-deoxycapsidiol + [reduced NADPH-hemoprotein reductase] + O2
capsidiol + [oxidized NADPH-hemoprotein reductase] + H2O
show the reaction diagram
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-
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-
?
3alpha-hydroxy-5-epiaristolochene + [reduced NADPH-hemoprotein reductase] + O2
capsidiol + [oxidized NADPH-hemoprotein reductase] + H2O
show the reaction diagram
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the catalytic efficiency for 3alpha-hydroxy-5-epiaristolochene is about 10times lower than that for 1beta-hydroxy-5-epiaristolochene
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?
5-epiaristolochene + [reduced NADPH-hemoprotein reductase] + O2
1-deoxycapsidiol + [oxidized NADPH-hemoprotein reductase] + H2O
show the reaction diagram
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-
-
-
?
5-epiaristolochene + [reduced NADPH-hemoprotein reductase] + O2
1beta-hydroxy-5-epiaristolochene + [oxidized NADPH-hemoprotein reductase] + H2O
show the reaction diagram
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the release of an 1beta-hydroxy-5-epiaristolochene intermediate at high 5-epiaristolochene concentrations and a 10fold catalytic preference for 1beta-hydroxy-5-epiaristolochene versus 3alpha-hydroxy-5-epiaristolochene is indicative of a preferred reaction order of hydroxylation at C-1, followed by that at C-3
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?
5-epiaristolochene + [reduced NADPH-hemoprotein reductase] + O2
3-deoxycapsidiol + [oxidized NADPH-hemoprotein reductase] + H2O
show the reaction diagram
5-epiaristolochene + [reduced NADPH-hemoprotein reductase] + O2
capsidiol + [oxidized NADPH-hemoprotein reductase] + H2O
show the reaction diagram
premnaspirodiene + [reduced NADPH-hemoprotein reductase] + O2
solavetivone + [oxidized NADPH-hemoprotein reductase] + H2O
show the reaction diagram
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?
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
1-deoxycapsidiol + [reduced NADPH-hemoprotein reductase] + O2
capsidiol + [oxidized NADPH-hemoprotein reductase] + H2O
show the reaction diagram
-
-
-
-
?
3-deoxycapsidiol + [reduced NADPH-hemoprotein reductase] + O2
capsidiol + [oxidized NADPH-hemoprotein reductase] + H2O
show the reaction diagram
-
-
-
-
?
5-epiaristolochene + [reduced NADPH-hemoprotein reductase] + O2
1-deoxycapsidiol + [oxidized NADPH-hemoprotein reductase] + H2O
show the reaction diagram
-
-
-
-
?
5-epiaristolochene + [reduced NADPH-hemoprotein reductase] + O2
3-deoxycapsidiol + [oxidized NADPH-hemoprotein reductase] + H2O
show the reaction diagram
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3-deoxycapsidiol is a reaction product when EAH is incubated with high concentrations (above 0.4 mM) of the 5-epiaristolochene substrate
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?
5-epiaristolochene + [reduced NADPH-hemoprotein reductase] + O2
capsidiol + [oxidized NADPH-hemoprotein reductase] + H2O
show the reaction diagram
-
-
-
-
?
premnaspirodiene + [reduced NADPH-hemoprotein reductase] + O2
solavetivone + [oxidized NADPH-hemoprotein reductase] + H2O
show the reaction diagram
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-
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-
?
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
Ancymidol
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dose-dependent inhibition with more than 80% by 0.075 mM ancymidol
dimethyl sulfoxide
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conversion of 5-epiaristolochene to capsidiol activity is inhibited at concentrations above 10% (v/v) dimethyl sulfoxide
ketoconazole
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dose-dependent inhibition with 95% inhibition at 0.1 mM ketoconazole
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
dimethyl sulfoxide
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maximum conversion of 5-epiaristolochene to capsidiol activity is observed at final concentrations of 2-5% (v/v) dimethyl sulfoxide
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.00174 - 0.02124
1beta-hydroxy-5-epiaristolochene
0.01518 - 0.0688
5-epiaristolochene
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.132 - 0.582
1beta-hydroxy-5-epiaristolochene
0.037 - 0.553
5-epiaristolochene
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
6.2 - 334.4
1beta-hydroxy-5-epiaristolochene
2.4 - 33.1
5-epiaristolochene
IC50 VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.025
Ancymidol
Nicotiana tabacum
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pH and temperature not specified in the publication
0.025
ketoconazole
Nicotiana tabacum
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pH and temperature not specified in the publication
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
expressed in a WAT11 line of yeast
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expressed in WAT11 yeast strain
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gene EAH, quantitative RT-PCR enzyme expression analysis, phylogenetic analysis
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EXPRESSION
ORGANISM
UNIPROT
LITERATURE
after an apparent lag phase of 8 h, a rapid induction of hydroxylase activity is observed 10 to 15 h after elicitor addition (cellulase or paraciticein) to the cell cultures, reaching a maximum by 18 h followed by a rather gradual decline of 10 to 20% over the next 8 h
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pathogen Phytophthora infestans isolates T30-4 and NL07434 highly induce the enzyme expression in Capsicum annuum, transcriptome analysis, overview
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ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
I468A
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the mutant produces significant amounts 1beta-hydroxy-5-epiaristolochene, but negligible amounts of capsidiol from 5-epiaristolochene
S368A
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the mutant retains its ability to fully convert 5-epiaristolochene to capsidiol, but the turnover rates for capsidiol formation is 3-13times lower compared with that of the wild type enzyme and is also able to convert 1beta-hydroxy-5-epiaristolochene to capsidiol with kcat values comparable with that of wild type enzyme, but with Km values for 1beta-hydroxy-5-epiaristolochene 4-6times greater compared with that of wild type
S368C
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the mutant exhibits wild type catalytic efficiency for 1beta-hydroxy-5-epiaristolochene biosynthesis, but is devoid of the successive hydroxylation activity for capsidiol biosynthesis
S368F
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devoid of any hydroxylase activity for 5-epiaristolochene or 1beta-hydroxy-5-epiaristolochene
S368I
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devoid of any hydroxylase activity for 5-epiaristolochene or 1beta-hydroxy-5-epiaristolochene
S368T
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the mutant retains its ability to fully convert 5-epiaristolochene to capsidiol, but the turnover rates for capsidiol formation is 3-13times lower compared with that of the wild type enzyme and is also able to convert 1beta-hydroxy-5-epiaristolochene to capsidiol with kcat values comparable with that of wild type enzyme, but with Km values for 1beta-hydroxy-5-epiaristolochene 4-6times greater compared with that of wild type
S368V
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the mutant exhibits wild type catalytic efficiency for 1beta-hydroxy-5-epiaristolochene biosynthesis, but is devoid of the successive hydroxylation activity for capsidiol biosynthesis, the mutant catalyzes the relative equal biosynthesis of 1beta-hydroxy-5-epiaristolochene, 2beta-hydroxy-5-epiaristolochene, and 3beta-hydroxy-5-epiaristolochene from 5-epiaristolochene with wild type efficiency and converts about 1.5% of these monohydroxylated products to their respective ketone forms