Information on EC 2.3.1.160 - vinorine synthase

Word Map on EC 2.3.1.160
Please wait a moment until all data is loaded. This message will disappear when all data is loaded.
Specify your search results
Select one or more organisms in this record:
Show additional data
Do not include text mining results
Include (text mining) results (more...)
Include results (AMENDA + additional results, but less precise; more...)


The expected taxonomic range for this enzyme is: Rauvolfia serpentina

EC NUMBER
COMMENTARY hide
2.3.1.160
-
RECOMMENDED NAME
GeneOntology No.
vinorine synthase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
acetyl-CoA + 16-epivellosimine = CoA + vinorine
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
cyclization
-
-
-
-
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
ajmaline and sarpagine biosynthesis
-
-
Biosynthesis of secondary metabolites
-
-
Indole alkaloid biosynthesis
-
-
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
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
Rauvolfia serpentina x Rhazya stricta
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
acetyl-CoA + gardneral
CoA + 11-methoxyvinorine
show the reaction diagram
additional information
?
-
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
acetyl-CoA + 16-epivellosimine
CoA + vinorine
show the reaction diagram
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
4-(2-aminoethyl)-benzenesulfonyl fluoride
Rauvolfia serpentina x Rhazya stricta
selective Ser modifying reagent, complete inhibition
diethyldicarbonate
Rauvolfia serpentina x Rhazya stricta
complete inhibition
Hg2+
Rauvolfia serpentina x Rhazya stricta
complete inhibition
N-(N-(L-3-trans-carboxirane-2-carbonyl)-L-leucyl)-agmantine
Rauvolfia serpentina x Rhazya stricta
i.e. E64, selective Cys modifying reagent, 50% inhibition
-
N-tosyl-L-phenylalanine chloromethylketone
Rauvolfia serpentina x Rhazya stricta
complete inhibition
Nalpha-p-tosyl-L-lysine chloromethylketone
Rauvolfia serpentina x Rhazya stricta
50% inhibition
PMSF
Rauvolfia serpentina x Rhazya stricta
58% inhibition
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.039
11-methoxy-16-epivellosimine
-
-
-
0.0194
16-epivellosimine
-
-
0.057 - 0.064
acetyl-CoA
0.063
CoA
Rauvolfia serpentina x Rhazya stricta
pH 7.0, 30C, recombinant wild-type enzyme
0.0011 - 0.027
gardneral
0.01
vinorine
Rauvolfia serpentina x Rhazya stricta
pH 7.0, 30C, recombinant wild-type enzyme
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.008 - 0.103
gardneral
0.735
vinorine
Rauvolfia serpentina x Rhazya stricta
Q70PR7
pH 7.0, 30C, recombinant wild-type enzyme
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
99.1
Rauvolfia serpentina x Rhazya stricta
-
purified enzyme
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
7
Rauvolfia serpentina x Rhazya stricta
-
assay at
7.8
Rauvolfia serpentina x Rhazya stricta
-
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
30
Rauvolfia serpentina x Rhazya stricta
-
assay at
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
pI VALUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5
Rauvolfia serpentina x Rhazya stricta
sequence calculation
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
Rauvolfia serpentina x Rhazya stricta
-
hybrid of Rauvolfia serpentina and Rhazya stricta
Manually annotated by BRENDA team
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
31000
-
gel filtration
42000
Rauvolfia serpentina x Rhazya stricta
-
about, gel filtration
46800
Rauvolfia serpentina x Rhazya stricta
1 x * 46800, wild-type untagged enzyme
50000
Rauvolfia serpentina x Rhazya stricta
-
1 * 50000, about, SDS-PAGE
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
monomer
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
purified detagged recombinant wild-type or selenomethionine-labeled enzyme, hanging drop vapour diffusion method, 32C, 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
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
991fold from cell suspension culture by anion exchange, hydrophobic interaction, and hydroxyapatite chromatography, and gel filtration
Rauvolfia serpentina x Rhazya stricta
-
of the recombinant protein by his-tag affinity column chromatography
-
recombinant N-terminally His-tagged wild-type and mutant enzymes from Escherichia coli to homogeneity
Rauvolfia serpentina x Rhazya stricta
recombinant soluble N-terminally His-tagged enzyme from Escherichia coli by nickel affinity chromatography and gel filtration
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
DNA sequence determination and analysis, functional overexpression of N-terminally His-tagged wild-type and mutant enzymes in Escherichia coli
Rauvolfia serpentina x Rhazya stricta
expression in Escherichia coli
-
overexpression of N-terminally His-tagged enzyme in Escherichia coli
ENGINEERING
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
C149A
Rauvolfia serpentina x Rhazya stricta
site-directed mutagenesis, 10% of wild-type activity
C89A
Rauvolfia serpentina x Rhazya stricta
site-directed mutagenesis, activity similar to the wild-type enzyme
D164A
Rauvolfia serpentina x Rhazya stricta
site-directed mutagenesis, inactive mutant
D32A
Rauvolfia serpentina x Rhazya stricta
site-directed mutagenesis, 14% of wild-type activity
D360A
Rauvolfia serpentina x Rhazya stricta
site-directed mutagenesis, activity similar to the wild-type enzyme
D362A
Rauvolfia serpentina x Rhazya stricta
site-directed mutagenesis, 35% of wild-type activity
H160A
Rauvolfia serpentina x Rhazya stricta
site-directed mutagenesis, inactive mutant
N293A
Rauvolfia serpentina x Rhazya stricta
site-directed mutagenesis, 68% of wild-type activity
S16A
Rauvolfia serpentina x Rhazya stricta
site-directed mutagenesis, 71% of wild-type activity
S243A
Rauvolfia serpentina x Rhazya stricta
site-directed mutagenesis, 17% of wild-type activity
S29A
Rauvolfia serpentina x Rhazya stricta
site-directed mutagenesis, 25% of wild-type activity
S413A
Rauvolfia serpentina x Rhazya stricta
site-directed mutagenesis, activity similar to the wild-type enzyme
S68A
Rauvolfia serpentina x Rhazya stricta
site-directed mutagenesis, activity similar to the wild-type enzyme