BRENDA - Enzyme Database show
show all sequences of 2.3.1.160

Acetyltransfer in natural product biosynthesis - functional cloning and molecular analysis of vinorine synthase

Bayer, A.; Ma, X.; Stockigt, J.; Bioorg. Med. Chem. 12, 2787-2795 (2004)

Data extracted from this reference:

Cloned(Commentary)
Commentary
Organism
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
Engineering
Amino acid exchange
Commentary
Organism
C149A
site-directed mutagenesis, 10% of wild-type activity
Rauvolfia serpentina x Rhazya stricta
C89A
site-directed mutagenesis, activity similar to the wild-type enzyme
Rauvolfia serpentina x Rhazya stricta
D164A
site-directed mutagenesis, inactive mutant
Rauvolfia serpentina x Rhazya stricta
D32A
site-directed mutagenesis, 14% of wild-type activity
Rauvolfia serpentina x Rhazya stricta
D360A
site-directed mutagenesis, activity similar to the wild-type enzyme
Rauvolfia serpentina x Rhazya stricta
D362A
site-directed mutagenesis, 35% of wild-type activity
Rauvolfia serpentina x Rhazya stricta
H160A
site-directed mutagenesis, inactive mutant
Rauvolfia serpentina x Rhazya stricta
N293A
site-directed mutagenesis, 68% of wild-type activity
Rauvolfia serpentina x Rhazya stricta
S16A
site-directed mutagenesis, 71% of wild-type activity
Rauvolfia serpentina x Rhazya stricta
S243A
site-directed mutagenesis, 17% of wild-type activity
Rauvolfia serpentina x Rhazya stricta
S29A
site-directed mutagenesis, 25% of wild-type activity
Rauvolfia serpentina x Rhazya stricta
S413A
site-directed mutagenesis, activity similar to the wild-type enzyme
Rauvolfia serpentina x Rhazya stricta
S68A
site-directed mutagenesis, activity similar to the wild-type enzyme
Rauvolfia serpentina x Rhazya stricta
Inhibitors
Inhibitors
Commentary
Organism
Structure
4-(2-aminoethyl)-benzenesulfonyl fluoride
selective Ser modifying reagent, complete inhibition
Rauvolfia serpentina x Rhazya stricta
diethyldicarbonate
complete inhibition
Rauvolfia serpentina x Rhazya stricta
Hg2+
complete inhibition
Rauvolfia serpentina x Rhazya stricta
N-(N-(L-3-trans-carboxirane-2-carbonyl)-L-leucyl)-agmantine
i.e. E64, selective Cys modifying reagent, 50% inhibition
Rauvolfia serpentina x Rhazya stricta
N-tosyl-L-phenylalanine chloromethylketone
complete inhibition
Rauvolfia serpentina x Rhazya stricta
Nalpha-p-tosyl-L-lysine chloromethylketone
50% inhibition
Rauvolfia serpentina x Rhazya stricta
PMSF
58% inhibition
Rauvolfia serpentina x Rhazya stricta
KM Value [mM]
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.0011
-
gardneral
pH 7.0, 30°C, recombinant mutant C149A
Rauvolfia serpentina x Rhazya stricta
0.0026
-
gardneral
pH 7.0, 30°C, recombinant mutant N293A
Rauvolfia serpentina x Rhazya stricta
0.0047
-
gardneral
pH 7.0, 30°C, recombinant mutant S29A
Rauvolfia serpentina x Rhazya stricta
0.0075
-
gardneral
pH 7.0, 30°C, recombinant wild-type enzyme
Rauvolfia serpentina x Rhazya stricta
0.01
-
vinorine
pH 7.0, 30°C, recombinant wild-type enzyme
Rauvolfia serpentina x Rhazya stricta
0.021
-
gardneral
pH 7.0, 30°C, recombinant mutant S413A
Rauvolfia serpentina x Rhazya stricta
0.027
-
gardneral
pH 7.0, 30°C, recombinant mutant C89A
Rauvolfia serpentina x Rhazya stricta
0.057
-
acetyl-CoA
pH 7.0, 30°C, recombinant wild-type enzyme
Rauvolfia serpentina x Rhazya stricta
0.063
-
CoA
pH 7.0, 30°C, recombinant wild-type enzyme
Rauvolfia serpentina x Rhazya stricta
Molecular Weight [Da]
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
46800
-
1 x * 46800, wild-type untagged enzyme
Rauvolfia serpentina x Rhazya stricta
Natural Substrates/ Products (Substrates)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
acetyl-CoA + 16-epivellosimine
Rauvolfia serpentina x Rhazya stricta
the enzyme is involved in a crucial step of antiarrhythmic drug ajmaline biosynthesis, pathway overview
CoA + vinorine
-
-
r
Organism
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
Rauvolfia serpentina x Rhazya stricta
Q70PR7
hybrid
-
Purification (Commentary)
Commentary
Organism
recombinant N-terminally His-tagged wild-type and mutant enzymes from Escherichia coli to homogeneity
Rauvolfia serpentina x Rhazya stricta
Reaction
Reaction
Commentary
Organism
acetyl-CoA + 16-epivellosimine = CoA + vinorine
His160 and Asp164 are catalytic site residues of the non-triad motif HxxxD
Rauvolfia serpentina x Rhazya stricta
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
acetyl-CoA + 16-epivellosimine
the enzyme is involved in a crucial step of antiarrhythmic drug ajmaline biosynthesis, pathway overview
661321
Rauvolfia serpentina x Rhazya stricta
CoA + vinorine
-
-
-
r
acetyl-CoA + 16-epivellosimine
substrate specificity in both reaction directions, the reverse reaction is preferred
661321
Rauvolfia serpentina x Rhazya stricta
CoA + vinorine
-
-
-
r
acetyl-CoA + gardneral
i.e. 11-methoxy-16-epivellosimine, sarpagan-type alkaloid
661321
Rauvolfia serpentina x Rhazya stricta
CoA + 11-methoxyvinorine
ajmalin-type alkaloid
-
-
r
additional information
no activity with polyneuridine, vomilenine, 17-O-acetyl-norajmaline, and N-methylgardneral
661321
Rauvolfia serpentina x Rhazya stricta
?
-
-
-
-
Subunits
Subunits
Commentary
Organism
monomer
1 x * 46800, wild-type untagged enzyme
Rauvolfia serpentina x Rhazya stricta
Temperature Optimum [°C]
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
35
-
-
Rauvolfia serpentina x Rhazya stricta
Turnover Number [1/s]
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
0.008
-
gardneral
pH 7.0, 30°C, recombinant mutant C149A
Rauvolfia serpentina x Rhazya stricta
0.018
-
gardneral
pH 7.0, 30°C, recombinant mutant S29A
Rauvolfia serpentina x Rhazya stricta
0.043
-
gardneral
pH 7.0, 30°C, recombinant mutant N293A
Rauvolfia serpentina x Rhazya stricta
0.065
0.103
gardneral
pH 7.0, 30°C, recombinant wild-type enzyme
Rauvolfia serpentina x Rhazya stricta
0.068
-
gardneral
pH 7.0, 30°C, recombinant mutant C89A
Rauvolfia serpentina x Rhazya stricta
0.085
-
gardneral
pH 7.0, 30°C, recombinant mutant S413A
Rauvolfia serpentina x Rhazya stricta
0.735
-
vinorine
pH 7.0, 30°C, recombinant wild-type enzyme
Rauvolfia serpentina x Rhazya stricta
pH Optimum
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
7.8
-
-
Rauvolfia serpentina x Rhazya stricta
pI Value
Organism
Commentary
pI Value Maximum
pI Value
Rauvolfia serpentina x Rhazya stricta
sequence calculation
-
5
Cloned(Commentary) (protein specific)
Commentary
Organism
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
Engineering (protein specific)
Amino acid exchange
Commentary
Organism
C149A
site-directed mutagenesis, 10% of wild-type activity
Rauvolfia serpentina x Rhazya stricta
C89A
site-directed mutagenesis, activity similar to the wild-type enzyme
Rauvolfia serpentina x Rhazya stricta
D164A
site-directed mutagenesis, inactive mutant
Rauvolfia serpentina x Rhazya stricta
D32A
site-directed mutagenesis, 14% of wild-type activity
Rauvolfia serpentina x Rhazya stricta
D360A
site-directed mutagenesis, activity similar to the wild-type enzyme
Rauvolfia serpentina x Rhazya stricta
D362A
site-directed mutagenesis, 35% of wild-type activity
Rauvolfia serpentina x Rhazya stricta
H160A
site-directed mutagenesis, inactive mutant
Rauvolfia serpentina x Rhazya stricta
N293A
site-directed mutagenesis, 68% of wild-type activity
Rauvolfia serpentina x Rhazya stricta
S16A
site-directed mutagenesis, 71% of wild-type activity
Rauvolfia serpentina x Rhazya stricta
S243A
site-directed mutagenesis, 17% of wild-type activity
Rauvolfia serpentina x Rhazya stricta
S29A
site-directed mutagenesis, 25% of wild-type activity
Rauvolfia serpentina x Rhazya stricta
S413A
site-directed mutagenesis, activity similar to the wild-type enzyme
Rauvolfia serpentina x Rhazya stricta
S68A
site-directed mutagenesis, activity similar to the wild-type enzyme
Rauvolfia serpentina x Rhazya stricta
Inhibitors (protein specific)
Inhibitors
Commentary
Organism
Structure
4-(2-aminoethyl)-benzenesulfonyl fluoride
selective Ser modifying reagent, complete inhibition
Rauvolfia serpentina x Rhazya stricta
diethyldicarbonate
complete inhibition
Rauvolfia serpentina x Rhazya stricta
Hg2+
complete inhibition
Rauvolfia serpentina x Rhazya stricta
N-(N-(L-3-trans-carboxirane-2-carbonyl)-L-leucyl)-agmantine
i.e. E64, selective Cys modifying reagent, 50% inhibition
Rauvolfia serpentina x Rhazya stricta
N-tosyl-L-phenylalanine chloromethylketone
complete inhibition
Rauvolfia serpentina x Rhazya stricta
Nalpha-p-tosyl-L-lysine chloromethylketone
50% inhibition
Rauvolfia serpentina x Rhazya stricta
PMSF
58% inhibition
Rauvolfia serpentina x Rhazya stricta
KM Value [mM] (protein specific)
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.0011
-
gardneral
pH 7.0, 30°C, recombinant mutant C149A
Rauvolfia serpentina x Rhazya stricta
0.0026
-
gardneral
pH 7.0, 30°C, recombinant mutant N293A
Rauvolfia serpentina x Rhazya stricta
0.0047
-
gardneral
pH 7.0, 30°C, recombinant mutant S29A
Rauvolfia serpentina x Rhazya stricta
0.0075
-
gardneral
pH 7.0, 30°C, recombinant wild-type enzyme
Rauvolfia serpentina x Rhazya stricta
0.01
-
vinorine
pH 7.0, 30°C, recombinant wild-type enzyme
Rauvolfia serpentina x Rhazya stricta
0.021
-
gardneral
pH 7.0, 30°C, recombinant mutant S413A
Rauvolfia serpentina x Rhazya stricta
0.027
-
gardneral
pH 7.0, 30°C, recombinant mutant C89A
Rauvolfia serpentina x Rhazya stricta
0.057
-
acetyl-CoA
pH 7.0, 30°C, recombinant wild-type enzyme
Rauvolfia serpentina x Rhazya stricta
0.063
-
CoA
pH 7.0, 30°C, recombinant wild-type enzyme
Rauvolfia serpentina x Rhazya stricta
Molecular Weight [Da] (protein specific)
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
46800
-
1 x * 46800, wild-type untagged enzyme
Rauvolfia serpentina x Rhazya stricta
Natural Substrates/ Products (Substrates) (protein specific)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
acetyl-CoA + 16-epivellosimine
Rauvolfia serpentina x Rhazya stricta
the enzyme is involved in a crucial step of antiarrhythmic drug ajmaline biosynthesis, pathway overview
CoA + vinorine
-
-
r
Purification (Commentary) (protein specific)
Commentary
Organism
recombinant N-terminally His-tagged wild-type and mutant enzymes from Escherichia coli to homogeneity
Rauvolfia serpentina x Rhazya stricta
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
acetyl-CoA + 16-epivellosimine
the enzyme is involved in a crucial step of antiarrhythmic drug ajmaline biosynthesis, pathway overview
661321
Rauvolfia serpentina x Rhazya stricta
CoA + vinorine
-
-
-
r
acetyl-CoA + 16-epivellosimine
substrate specificity in both reaction directions, the reverse reaction is preferred
661321
Rauvolfia serpentina x Rhazya stricta
CoA + vinorine
-
-
-
r
acetyl-CoA + gardneral
i.e. 11-methoxy-16-epivellosimine, sarpagan-type alkaloid
661321
Rauvolfia serpentina x Rhazya stricta
CoA + 11-methoxyvinorine
ajmalin-type alkaloid
-
-
r
additional information
no activity with polyneuridine, vomilenine, 17-O-acetyl-norajmaline, and N-methylgardneral
661321
Rauvolfia serpentina x Rhazya stricta
?
-
-
-
-
Subunits (protein specific)
Subunits
Commentary
Organism
monomer
1 x * 46800, wild-type untagged enzyme
Rauvolfia serpentina x Rhazya stricta
Temperature Optimum [°C] (protein specific)
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
35
-
-
Rauvolfia serpentina x Rhazya stricta
Turnover Number [1/s] (protein specific)
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
0.008
-
gardneral
pH 7.0, 30°C, recombinant mutant C149A
Rauvolfia serpentina x Rhazya stricta
0.018
-
gardneral
pH 7.0, 30°C, recombinant mutant S29A
Rauvolfia serpentina x Rhazya stricta
0.043
-
gardneral
pH 7.0, 30°C, recombinant mutant N293A
Rauvolfia serpentina x Rhazya stricta
0.065
0.103
gardneral
pH 7.0, 30°C, recombinant wild-type enzyme
Rauvolfia serpentina x Rhazya stricta
0.068
-
gardneral
pH 7.0, 30°C, recombinant mutant C89A
Rauvolfia serpentina x Rhazya stricta
0.085
-
gardneral
pH 7.0, 30°C, recombinant mutant S413A
Rauvolfia serpentina x Rhazya stricta
0.735
-
vinorine
pH 7.0, 30°C, recombinant wild-type enzyme
Rauvolfia serpentina x Rhazya stricta
pH Optimum (protein specific)
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
7.8
-
-
Rauvolfia serpentina x Rhazya stricta
pI Value (protein specific)
Organism
Commentary
pI Value Maximum
pI Value
Rauvolfia serpentina x Rhazya stricta
sequence calculation
-
5
Other publictions for EC 2.3.1.160
No.
1st author
Pub Med
title
organims
journal
volume
pages
year
Activating Compound
Application
Cloned(Commentary)
Crystallization (Commentary)
Engineering
General Stability
Inhibitors
KM Value [mM]
Localization
Metals/Ions
Molecular Weight [Da]
Natural Substrates/ Products (Substrates)
Organic Solvent Stability
Organism
Oxidation Stability
Posttranslational Modification
Purification (Commentary)
Reaction
Renatured (Commentary)
Source Tissue
Specific Activity [micromol/min/mg]
Storage Stability
Substrates and Products (Substrate)
Subunits
Temperature Optimum [°C]
Temperature Range [°C]
Temperature Stability [°C]
Turnover Number [1/s]
pH Optimum
pH Range
pH Stability
Cofactor
Ki Value [mM]
pI Value
IC50 Value
Activating Compound (protein specific)
Application (protein specific)
Cloned(Commentary) (protein specific)
Cofactor (protein specific)
Crystallization (Commentary) (protein specific)
Engineering (protein specific)
General Stability (protein specific)
IC50 Value (protein specific)
Inhibitors (protein specific)
Ki Value [mM] (protein specific)
KM Value [mM] (protein specific)
Localization (protein specific)
Metals/Ions (protein specific)
Molecular Weight [Da] (protein specific)
Natural Substrates/ Products (Substrates) (protein specific)
Organic Solvent Stability (protein specific)
Oxidation Stability (protein specific)
Posttranslational Modification (protein specific)
Purification (Commentary) (protein specific)
Renatured (Commentary) (protein specific)
Source Tissue (protein specific)
Specific Activity [micromol/min/mg] (protein specific)
Storage Stability (protein specific)
Substrates and Products (Substrate) (protein specific)
Subunits (protein specific)
Temperature Optimum [°C] (protein specific)
Temperature Range [°C] (protein specific)
Temperature Stability [°C] (protein specific)
Turnover Number [1/s] (protein specific)
pH Optimum (protein specific)
pH Range (protein specific)
pH Stability (protein specific)
pI Value (protein specific)
Expression
General Information
General Information (protein specific)
Expression (protein specific)
KCat/KM [mM/s]
KCat/KM [mM/s] (protein specific)
682286
Stoeckigt
-
The molecular architecture of ...
Rauvolfia serpentina
Phytochem. Rev.
6
15-34
2007
-
-
1
1
5
-
-
-
-
-
-
-
-
1
-
-
1
-
-
-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
1
5
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
662350
Ma
Crystal structure of vinorine ...
Rauvolfia serpentina
J. Biol. Chem.
280
13576-13583
2005
-
-
1
1
-
-
-
-
-
-
-
1
-
3
-
-
1
1
-
-
-
-
2
1
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
1
-
-
-
-
-
-
-
-
-
1
-
-
-
1
-
-
-
-
2
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
661320
Gerasimenko
Purification and partial amino ...
Rauvolfia serpentina x Rhazya stricta
Bioorg. Med. Chem.
12
2781-2786
2004
-
-
-
-
-
-
-
2
-
-
2
1
-
5
-
-
1
-
-
3
1
-
4
1
1
-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
2
-
-
2
1
-
-
-
1
-
3
1
-
4
1
1
-
-
-
1
-
-
-
-
-
-
-
-
-
661321
Bayer
Acetyltransfer in natural prod ...
Rauvolfia serpentina x Rhazya stricta
Bioorg. Med. Chem.
12
2787-2795
2004
-
-
1
-
13
-
7
9
-
-
1
1
-
7
-
-
1
1
-
-
-
-
4
1
1
-
-
7
1
-
-
-
-
1
-
-
-
1
-
-
13
-
-
7
-
9
-
-
1
1
-
-
-
1
-
-
-
-
4
1
1
-
-
7
1
-
-
1
-
-
-
-
-
-
486439
Pfitzner
-
Properties of vinorine synthas ...
Rauvolfia serpentina
Z. Naturforsch. C
41
103-114
1986
-
-
-
-
-
-
-
3
-
-
1
1
-
1
-
-
1
-
-
-
1
-
2
-
1
1
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
3
-
-
1
1
-
-
-
1
-
-
1
-
2
-
1
1
-
-
1
-
-
-
-
-
-
-
-
-
486440
Pfitzner
-
Biogenetic link between sarpag ...
Rauvolfia serpentina
Tetrahedron Lett.
24
5197-5200
1983
-
-
-
-
-
-
-
-
-
-
-
1
-
1
-
-
1
-
-
-
-
-
1
-
-
-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
-
1
-
-
-
-
1
-
-
-
-
-
1
-
-
-
-
-
-
-
-
-