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Literature summary for 2.3.1.160 extracted from

  • Bayer, A.; Ma, X.; Stockigt, J.
    Acetyltransfer in natural product biosynthesis - functional cloning and molecular analysis of vinorine synthase (2004), Bioorg. Med. Chem., 12, 2787-2795.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) 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

Protein Variants

Protein Variants Comment 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 Comment 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)-agmatine 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 Comment 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] Comment Organism
46800
-
1 x * 46800, wild-type untagged enzyme Rauvolfia serpentina x Rhazya stricta

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
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 UniProt Comment Textmining
Rauvolfia serpentina x Rhazya stricta Q70PR7 hybrid
-

Purification (Commentary)

Purification (Comment) Organism
recombinant N-terminally His-tagged wild-type and mutant enzymes from Escherichia coli to homogeneity Rauvolfia serpentina x Rhazya stricta

Reaction

Reaction Comment Organism Reaction ID
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 Comment Substrates Organism Products Comment (Products) Rev. Reac.
acetyl-CoA + 16-epivellosimine the enzyme is involved in a crucial step of antiarrhythmic drug ajmaline biosynthesis, pathway overview Rauvolfia serpentina x Rhazya stricta CoA + vinorine
-
r
acetyl-CoA + 16-epivellosimine substrate specificity in both reaction directions, the reverse reaction is preferred Rauvolfia serpentina x Rhazya stricta CoA + vinorine
-
r
acetyl-CoA + gardneral i.e. 11-methoxy-16-epivellosimine, sarpagan-type alkaloid 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 Rauvolfia serpentina x Rhazya stricta ?
-
?

Subunits

Subunits Comment Organism
monomer 1 x * 46800, wild-type untagged enzyme Rauvolfia serpentina x Rhazya stricta

Synonyms

Synonyms Comment Organism
More enzyme belongs to the BAHD acetyltransferase super family Rauvolfia serpentina x Rhazya stricta
vinorine synthase
-
Rauvolfia serpentina x Rhazya stricta
VS
-
Rauvolfia serpentina x Rhazya stricta

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
35
-
-
Rauvolfia serpentina x Rhazya stricta

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment 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 Comment Organism
7.8
-
-
Rauvolfia serpentina x Rhazya stricta

pI Value

Organism Comment pI Value Maximum pI Value
Rauvolfia serpentina x Rhazya stricta sequence calculation
-
5