Information on EC 1.14.11.14 - 6beta-hydroxyhyoscyamine epoxidase

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The enzyme appears in viruses and cellular organisms

EC NUMBER
COMMENTARY hide
1.14.11.14
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RECOMMENDED NAME
GeneOntology No.
6beta-hydroxyhyoscyamine epoxidase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
(6S)-6beta-hydroxyhyoscyamine + 2-oxoglutarate + O2 = scopolamine + succinate + CO2 + H2O
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
oxidation
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redox reaction
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-
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reduction
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-
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PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
hyoscyamine and scopolamine biosynthesis
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superpathway of hyoscyamine and scopolamine biosynthesis
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Tropane, piperidine and pyridine alkaloid biosynthesis
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Metabolic pathways
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Biosynthesis of secondary metabolites
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SYSTEMATIC NAME
IUBMB Comments
(6S)-6beta-hydroxyhyoscyamine,2-oxoglutarate oxidoreductase (epoxide-forming)
Requires Fe2+ and ascorbate.
CAS REGISTRY NUMBER
COMMENTARY hide
121479-53-6
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ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
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Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
evolution
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the enzyme is a member of the 2-oxoglutarate-dependent dioxygenase family
metabolism
additional information
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
(6S)-6-hydroxyhyoscyamine + 2-oxoglutarate + O2
scopolamine + succinate + CO2
show the reaction diagram
(6S)-6beta-hydroxyhyoscyamine + 2-oxoglutarate + O2
scopolamine + succinate + CO2
show the reaction diagram
(6S)-hydroxyhyoscyamine + 2-oxoglutarate + O2
scopolamine + succinate + CO2
show the reaction diagram
-
-
-
?
6,7-dehydrohyoscyamine + 2-oxoglutarate + O2
?
show the reaction diagram
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-
-
-
?
8-methyl-8-azabicyclo[3.2.1]octan-3-yl 2-(4-(dimethylamino)phenyl)acetate + 2-oxoglutarate + O2
?
show the reaction diagram
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-
-
-
?
8-methyl-8-azabicyclo[3.2.1]octan-3-yl benzoate + 2-oxoglutarate + O2
?
show the reaction diagram
-
-
-
-
?
additional information
?
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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
(6S)-6-hydroxyhyoscyamine + 2-oxoglutarate + O2
scopolamine + succinate + CO2
show the reaction diagram
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alkaloid metabolism, scopolamine is formed by oxidative transformation of hyoscyamine in several solanaceous species
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?
(6S)-6beta-hydroxyhyoscyamine + 2-oxoglutarate + O2
scopolamine + succinate + CO2
show the reaction diagram
(6S)-hydroxyhyoscyamine + 2-oxoglutarate + O2
scopolamine + succinate + CO2
show the reaction diagram
A2IBI0
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?
additional information
?
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hyoscyamine 6beta-hydroxylase, a 2-oxoglutarate-dependent dioxygenase, converts L-hyoscyamine to its 6,7-epoxy derivative, scopolamine, in two sequential steps, the enzyme also catalyzes the reaaction of EC 1.14.11.11. 6,7-Dehydrohyoscyamine, a potential precursor for the last step of epoxidation, is an obligatory intermediate in the biosynthesis of scopolamine
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COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2-oxoglutarate
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ascorbate
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METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
(1R,2R,4S,5S,7s)-7-{[(2S)-3-hydroxy-2-phenylpropanoyl]oxy}-9,9-dimethyl-3-oxa-9-azoniatricyclo[3.3.1.02,4]nonane
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; slight inhibition by the substrate analogue
1-methylpiperidin-4-yl 2-phenylacetate
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6,7-Dehydrohyoscyamine
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8-methyl-8-azabicyclo[3.2.1]octan-3-yl 2-(4-(dimethylamino)phenyl)acetate
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8-methyl-8-azabicyclo[3.2.1]octan-3-yl 3-phenylpropanoate
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8-methyl-8-azabicyclo[3.2.1]octan-3-yl benzoate
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8-oxabicyclo[3.2.1]octan-3-yl 2-phenylacetate
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CuSO4
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100% inhibition
MgSO4
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24% inhibition
MnSO4
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100% inhibition
Pyridine 2,4-dicarboxylate
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ZnSO4
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100% inhibition
additional information
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ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
ascorbate
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required for activity
catalase
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KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.015
(6S)-6-hydroxyhyoscyamine
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0.0844
(6S)-6beta-hydroxyhyoscyamine
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pH 7.6, 34C, recombinant enzyme
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.000198
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pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
7.6
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recombinant enzyme
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
34
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recombinant enzyme
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
?
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x * 64000, recombinant GST-tagged enzyme, SDS-PAGE, x * 43000, recombinant His-tagged enzyme, SDS-PAGE
monomer
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1 * 39238, recombinant enzyme, mass spectrometry, 1 * 39369, sequence calculation
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
rapid loss of enzyme activity during purification
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OXIDATION STABILITY
ORGANISM
UNIPROT
LITERATURE
requires molecular oxygen
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439268
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-20C, enzyme is stored in small portions until assayed
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-20C, purified recombinant enzyme, 20 mM phosphate, 2 months, loss of approximately 20% of activity
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4C, purified recombinant enzyme, 20 mM phosphate, 4 days, loss of over 50% activity
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Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
partial, could not be separated from hyoscyamine 6beta-hydroxylase
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recombinant N- or C-terminally His-tagged from Escherichia coli by nickel affinity chromatography
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recombinant N-terminally His- or GST-tagged enzyme from Escherichia coli strain BL21 (DE3) by nickel or glutathione affinity chromatography, respectively
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Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
DNA and amino acid sequence determination and analysis, overexpression of N- or C-terminally His-tagged enzyme in Escherichia coli strain BL21 (DE3), AbH6H loses of the first amino acid methionine during transcription
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DNA and amino acid sequence determination and analysis, overexpression of N-terminally His- or GST-tagged enzyme in Escherichia coli strain BL21 (DE3)
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overexpression of the enzyme in hairy roots of transgenic Anisodus acutangulus using Agrobacterium tumefaciens strain C58C1 transfection method, coexpression with tropinone reductase I; overexpression of the enzyme in hairy roots of transgenic Anisodus acutangulus using Agrobacterium tumefaciens strain C58C1 transfection method, coexpression with tropinone reductase I
EXPRESSION
ORGANISM
UNIPROT
LITERATURE
trivalent chromium, as KCr(SO4)2, induces the enzyme expression at 1 mM
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ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
enhancied production of tropane alkaloids in transgenic Anisodus acutangulus hairy root cultures by overexpressing tropinone reductase I and hyoscyamine-6beta-hydroxylase Agrobacterium-mediated gene transfer technology, quantification of tropanalkaloid contents in transgenic lines, overview; enhancied production of tropane alkaloids in transgenic Anisodus acutangulus hairy root cultures by overexpressing tropinone reductase I and hyoscyamine-6beta-hydroxylase, quantification of tropan alkaloid contents in transgenic lines, overview
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
pharmacology
tropane alkaloids including hyoscyamine, anisodamine, scopolamine and anisodine, are used medicinally as anticholinergic agents with increasing market demand, improvement of production by metabolic engineering introduction of genes encoding the branch-controlling enzyme tropinone reductase I and the downstream rate-limiting enzyme hyoscyamine-6beta-hydroxylase; tropane alkaloids including hyoscyamine, anisodamine, scopolamine and anisodine, are used medicinally as anticholinergic agents with increasing market demand, improvement of production by metabolic engineering introduction of genes encoding the branch-controlling enzyme tropinone reductase I and the downstream rate-limiting enzyme hyoscyamine-6beta-hydroxylase