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Literature summary for 1.14.11.11 extracted from

  • Li, J.; van Belkum, M.J.; Vederas, J.C.
    Functional characterization of recombinant hyoscyamine 6beta-hydroxylase from Atropa belladonna (2012), Bioorg. Med. Chem., 20, 4356-4363.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
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 Atropa belladonna

Inhibitors

Inhibitors Comment Organism Structure
(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 slight inhibition by the substrate analogue Atropa belladonna
1-methylpiperidin-4-yl 2-phenylacetate
-
Atropa belladonna
6,7-Dehydrohyoscyamine
-
Atropa belladonna
8-methyl-8-azabicyclo[3.2.1]octan-3-yl 2-(4-(dimethylamino)phenyl)acetate
-
Atropa belladonna
8-methyl-8-azabicyclo[3.2.1]octan-3-yl 3-phenylpropanoate
-
Atropa belladonna
8-methyl-8-azabicyclo[3.2.1]octan-3-yl benzoate
-
Atropa belladonna
8-oxabicyclo[3.2.1]octan-3-yl 2-phenylacetate
-
Atropa belladonna
additional information synthesis and inhibitory potencies of substrate analogues, overview. No inhibition by substrate analogues 1-methylpiperidin-4-yl 2-phenylacetate, 8-methyl-8-azabicyclo[3.2.1]octan-3-yl 3-phenylpropanoate, and 8-oxabicyclo[3.2.1]octan-3-yl 2-phenylacetate Atropa belladonna

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.0521
-
L-hyoscyamine pH 7.6, 34°C, recombinant enzyme Atropa belladonna

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
39238
-
1 * 39238, recombinant enzyme, mass spectrometry, 1 * 39369, sequence calculation Atropa belladonna
39369
-
1 * 39238, recombinant enzyme, mass spectrometry, 1 * 39369, sequence calculation Atropa belladonna

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
L-hyoscyamine + 2-oxoglutarate + O2 Atropa belladonna
-
(6S)-hydroxyhyoscyamine + succinate + CO2
-
?
additional information Atropa belladonna hyoscyamine 6beta-hydroxylase, a 2-oxoglutarate-dependent dioxygenase, converts L-hyoscyamine to its 6,7-epoxy derivative, scopolamine, in two sequential steps. 6,7-Dehydrohyoscyamine, a potential precursor for the last step of epoxidation, is an obligatory intermediate in the biosynthesis of scopolamine ?
-
?

Organism

Organism UniProt Comment Textmining
Atropa belladonna
-
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant N- or C-terminally His-tagged from Escherichia coli by nickel affinity chromatography Atropa belladonna

Reaction

Reaction Comment Organism Reaction ID
L-hyoscyamine + 2-oxoglutarate + O2 = (6S)-hydroxyhyoscyamine + succinate + CO2 proposed mechanism of epoxidation catalyzed by the enzyme, overview Atropa belladonna

Storage Stability

Storage Stability Organism
-20°C, purified recombinant enzyme, 20 mM phosphate, 2 months, loss of approximately 20% of activity Atropa belladonna
4°C, purified recombinant enzyme, 20 mM phosphate, 4 days, loss of over 50% activity Atropa belladonna

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
8-methyl-8-azabicyclo[3.2.1]octan-3-yl 2-(4-(dimethylamino)phenyl)acetate + 2-oxoglutarate + O2
-
Atropa belladonna ?
-
?
8-methyl-8-azabicyclo[3.2.1]octan-3-yl benzoate + 2-oxoglutarate + O2
-
Atropa belladonna ?
-
?
L-hyoscyamine + 2-oxoglutarate + O2
-
Atropa belladonna (6S)-hydroxyhyoscyamine + succinate + CO2
-
?
additional information hyoscyamine 6beta-hydroxylase, a 2-oxoglutarate-dependent dioxygenase, converts L-hyoscyamine to its 6,7-epoxy derivative, scopolamine, in two sequential steps. 6,7-Dehydrohyoscyamine, a potential precursor for the last step of epoxidation, is an obligatory intermediate in the biosynthesis of scopolamine Atropa belladonna ?
-
?
additional information hyoscyamine 6beta-hydroxylase, a 2-oxoglutarate-dependent dioxygenase, converts L-hyoscyamine to its 6,7-epoxy derivative, scopolamine, in two sequential steps. The catalytic efficiency of AbH6H, especially for the second oxidation, which is the reaction of EC 1.14.11.14, is low. The epoxidation step is much slower than the hydroxylation step. Substrate analogues 1-methylpiperidin-4-yl 2-phenylacetate, 8-methyl-8-azabicyclo[3.2.1]octan-3-yl 3-phenylpropanoate, 8-oxabicyclo[3.2.1]octan-3-yl 2-phenylacetate, and 4 are no substrates for the enzyme Atropa belladonna ?
-
?

Subunits

Subunits Comment Organism
monomer 1 * 39238, recombinant enzyme, mass spectrometry, 1 * 39369, sequence calculation Atropa belladonna

Synonyms

Synonyms Comment Organism
H6H
-
Atropa belladonna
hyoscyamine 6beta-hydroxylase
-
Atropa belladonna

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
34
-
recombinant enzyme Atropa belladonna

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.6
-
recombinant enzyme Atropa belladonna

General Information

General Information Comment Organism
additional information the nitrogen atom in the tropane ring of L-hyoscyamine plays an important role in substrate recognition. Proposed mechanism of epoxidation catalyzed by H6H, overview Atropa belladonna