AUTHOR=Farzana Maisha , Richardson Matthew Bailey , Deschênes Daniel André Ramey , Mai Zhan , Njoku Destiny Ichechi , Deslongchamps Ghislain , Qu Yang 

TITLE=Parallel evolution of methyltransferases leads to vobasine biosynthesis in Tabernaemontana elegans and Catharanthus roseus

JOURNAL=Frontiers in Plant Science

VOLUME=15

YEAR=2024

URL=https://www.frontiersin.org/journals/plant-science/articles/10.3389/fpls.2024.1451298

DOI=10.3389/fpls.2024.1451298

ISSN=1664-462X

ABSTRACT=<p>Monoterpenoid indole alkaloids (MIA) are one of the largest and most complex alkaloid class in nature, boasting many clinically significant drugs such as anticancer vinblastine and antiarrhythmic ajmaline. Many MIAs undergo nitrogen <italic>N</italic>-methylation, altering their reactivity and affinity to the biological targets through a straightforward reaction. Remarkably, all known MIA <italic>N</italic>-methyltransferases (NMT) originate from the neofunctionalization of ancestral γ-tocopherol <italic>C</italic>-methyltransferases (γTMTs), a phenomenon seemingly unique to the Apocynaceae family. In this study, we unveil and characterize a new γTMT-like enzyme from the plant <italic>Tabernaemontana elegans</italic> (toad tree): perivine <italic>N</italic>β-methyltransferase (TePeNMT). TePeNMT and other homologs form a distinct clade in our phylogenetic study, setting them apart from other γTMTs and γTMT-like NMTs discovered to date. Enzyme kinetic experiments and enzyme homology modeling studies reveal the significant differences in enzyme active sites between TePeNMT and CrPeNMT, a previously characterized perivine <italic>N</italic>β-methyltransferase from <italic>Catharanthus roseus</italic> (Madagascar periwinkle). Collectively, our findings suggest that parallel evolution of ancestral γTMTs may be responsible for the occurrence of perivine <italic>N</italic>-methylation in <italic>T. elegans</italic> and <italic>C. roseus.</italic></p>