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 | 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 | 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 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 Maximum [Da] | Comment | Organism |
---|---|---|---|
46800 | - |
1 x * 46800, wild-type untagged enzyme | Rauvolfia serpentina x Rhazya stricta |
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 | UniProt | Comment | Textmining |
---|---|---|---|
Rauvolfia serpentina x Rhazya stricta | Q70PR7 | hybrid | - |
Purification (Comment) | Organism |
---|---|
recombinant N-terminally His-tagged wild-type and mutant enzymes from Escherichia coli to homogeneity | Rauvolfia serpentina x Rhazya stricta |
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 | 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 | Comment | Organism |
---|---|---|
monomer | 1 x * 46800, wild-type untagged enzyme | Rauvolfia serpentina x Rhazya stricta |
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 Maximum [°C] | Comment | Organism |
---|---|---|---|
35 | - |
- |
Rauvolfia serpentina x Rhazya stricta |
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 Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.8 | - |
- |
Rauvolfia serpentina x Rhazya stricta |
Organism | Comment | pI Value Maximum | pI Value |
---|---|---|---|
Rauvolfia serpentina x Rhazya stricta | sequence calculation | - |
5 |