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Information on EC 2.3.1.160 - vinorine synthase and Organism(s) Rauvolfia serpentina and UniProt Accession Q70PR7

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EC Tree
     2 Transferases
         2.3 Acyltransferases
             2.3.1 Transferring groups other than aminoacyl groups
                2.3.1.160 vinorine synthase
IUBMB Comments
The reaction proceeds in two stages. The indole nitrogen of 16-epivellosimine interacts with its aldehyde group giving an hydroxy-substituted new ring. This alcohol is then acetylated. Also acts on gardneral (11-methoxy-16-epivellosimine). Generates the ajmalan skeleton, which forms part of the route to ajmaline.
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This record set is specific for:
Rauvolfia serpentina
UNIPROT: Q70PR7
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Word Map
The taxonomic range for the selected organisms is: Rauvolfia serpentina
The expected taxonomic range for this enzyme is: Eukaryota, Bacteria
Synonyms
vinorine synthase, more
SYNONYM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
vinorine synthase
additional information
enzyme belongs to the BAHD enzyme super family
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
acetyl-CoA + 16-epivellosimine = CoA + vinorine
show the reaction diagram
His160 of the HxxxD motif functions as a general base in catalysis, located at the center of the reaction channel at the interface of both domains and is accessible from both sides
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
cyclization
-
-
-
-
SYSTEMATIC NAME
IUBMB Comments
acyl-CoA:16-epivellosimine O-acetyltransferase (cyclizing)
The reaction proceeds in two stages. The indole nitrogen of 16-epivellosimine interacts with its aldehyde group giving an hydroxy-substituted new ring. This alcohol is then acetylated. Also acts on gardneral (11-methoxy-16-epivellosimine). Generates the ajmalan skeleton, which forms part of the route to ajmaline.
CAS REGISTRY NUMBER
COMMENTARY hide
88844-97-7
-
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
11-methoxy-16-epivellosimine + acetyl-CoA
CoA + 11-methoxy-vinorine
show the reaction diagram
-
-
-
-
?
16-epi-vellosimine + acetyl-CoA
vinorine + CoA
show the reaction diagram
-
VS responsible for generation of the basic carbon skeleton of the target compound ajmaline, His160 and Asp164 are the most important amino acids for the catalytic process, His-X-X-X-Asp sequence together with Asp-Phe-Gly-Trp-Gly are highly conserved motifs in acyltransferases
-
-
?
acetyl-CoA + 16-epivellosimine
CoA + vinorine
show the reaction diagram
NATURAL SUBSTRATE
NATURAL PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
acetyl-CoA + 16-epivellosimine
CoA + vinorine
show the reaction diagram
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.039
11-methoxy-16-epivellosimine
-
-
0.0194
16-epivellosimine
-
-
0.064
acetyl-CoA
-
-
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
UNIPROT
ENTRY NAME
ORGANISM
NO. OF AA
NO. OF TRANSM. HELICES
MOLECULAR WEIGHT[Da]
SOURCE
SEQUENCE
LOCALIZATION PREDICTION?
VINSY_RAUSE
421
0
46828
Swiss-Prot
other Location (Reliability: 2)
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
31000
-
gel filtration
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
monomer
two-domain structure
CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
purified detagged recombinant wild-type or selenomethionine-labeled enzyme, hanging drop vapour diffusion method, 32°C, 2-3 mg/ml protein in 20 mM Tris-HCl, pH 7.5, 10 mM 2-mercaptoethanol, 1 mM EDTA, 0.5 mM acetyl-CoA, with reservoir solution containing 0.1 M Tris-HCl, pH 8.7, 2 M ammonium sulfate, 2% PEG 400, cryoprotection by 20-25% glycerol, X-ray diffraction structure determination anad analysis at 2.6 A resolution, modeling of CoA binding
using the hanging-drop vapour-diffusion method
-
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
D164A
-
reduction in enzyme activity
D32A
-
reduction in enzyme activity
D362A
-
reduction in enzyme activity
H160A
-
reduction in enzyme activity
S29A
-
reduction in enzyme activity
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
of the recombinant protein by his-tag affinity column chromatography
-
recombinant soluble N-terminally His-tagged enzyme from Escherichia coli by nickel affinity chromatography and gel filtration
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
expression in Escherichia coli
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overexpression of N-terminally His-tagged enzyme in Escherichia coli
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Pfitzner, A.; Polz, L.; Stckligt, J.
Properties of vinorine synthase the Rauwolfia enzyme involved in the formation of the ajmaline skeleton
Z. Naturforsch. C
41
103-114
1986
Rauvolfia serpentina
-
Manually annotated by BRENDA team
Pfitzner, A.; Stoeckigt, J.
Biogenetic link between sarpagine and ajmaline type alkaloids
Tetrahedron Lett.
24
5197-5200
1983
Rauvolfia serpentina
-
Manually annotated by BRENDA team
Ma, X.; Koepke, J.; Panjikar, S.; Fritzsch, G.; Stockigt, J.
Crystal structure of vinorine synthase, the first representative of the BAHD superfamily
J. Biol. Chem.
280
13576-13583
2005
Rauvolfia serpentina (Q70PR7)
Manually annotated by BRENDA team
Stoeckigt, J.; Panjikar, S.; Ruppert, M.; Barleben, L.; Ma, X.; Loris, E.; Hill, M.
The molecular architecture of major enzymes from ajmaline biosynthetic pathway
Phytochem. Rev.
6
15-34
2007
Rauvolfia serpentina
-
Manually annotated by BRENDA team