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Rauvolfia serpentina N‐methyltransferases involved in ajmaline and Nβ‐methylajmaline biosynthesis belong to a gene family derived from γ‐tocopherol C‐methyltransferase
- Cázares‐Flores, Paulo, Levac, Dylan, De Luca, Vincenzo
- The plant journal 2016 v.87 no.4 pp. 335-342
- Escherichia coli, Rauvolfia serpentina, biochemical pathways, biosynthesis, enzyme activity, gamma-tocopherol, genes, indole alkaloids, messenger RNA, molecular cloning, monoterpenoids, recombinant proteins, roots, substrate specificity
- Ajmaline biosynthesis in Rauvolfia serpentina has been one of the most studied monoterpenoid indole alkaloid (MIA) pathways within the plant family Apocynaceae. Detailed molecular and biochemical information on most of the steps involved in the pathway has been generated over the last 30 years. Here we report the identification, molecular cloning and functional expression in Escherichia coli of two R. serpentinacDNAs that are part of a recently discovered γ‐tocopherol‐like N‐methyltransferase (γ‐TLMT) family and are involved in indole and side‐chain N‐methylation of ajmaline. Recombinant proteins showed remarkable substrate specificity for molecules with an ajmalan‐type backbone and strict regiospecific N‐methylation. Furthermore, N‐methyltransferase gene transcripts and enzyme activity were enriched in R. serpentina roots which correlated with accumulation of ajmaline alkaloid. This study elucidates the final step in the ajmaline biosynthetic pathway and describes the enzyme responsible for the formation of Nᵦ‐methylajmaline, an unusual charged MIA found in R. serpentina.