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Carqueijeiro I , Duge de Bernonville T , Lanoue A , Dang TT , Teijaro C , Paetz C , Billet K , Mosquera A , Oudin A , Besseau S , Papon N , Glevarec G , Atehortua L , Clastre M , Giglioli-Guivarc'h N , Schneider B , St-Pierre B , Andrade RB , O'Connor SE , Courdavault V
A BAHD acyltransferase catalyzing 19-O-acetylation of tabersonine derivatives in roots of Catharanthus roseus enables combinatorial synthesis of monoterpene indole alkaloids
Plant J. 2018 May;94(3) :469-484
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While the characterization of the biosynthetic pathway of monoterpene indole alkaloids (MIAs) in leaves of Catharanthus roseus is now reaching completion, only two enzymes from the root counterpart dedicated to tabersonine metabolism have been identified to date, namely tabersonine 19-hydroxylase (T19H) and minovincine 19-O-acetyltransferase (MAT). Albeit the recombinant MAT catalyzes MIA acetylation at low efficiency in vitro, we demonstrated that MAT was inactive when expressed in yeast and in planta, suggesting an alternative function for this enzyme. Therefore, through transcriptomic analysis of periwinkle adventitious roots, several other BAHD acyltransferase candidates were identified based on the correlation of their expression profile with T19H and found to localize in small genomic clusters. Only one, named tabersonine derivative 19-O-acetyltransferase (TAT) was able to acetylate the 19-hydroxytabersonine derivatives from roots, such as minovincinine and horhammericine, following expression in yeast. Kinetic studies also showed that the recombinant TAT was specific for root MIAs and displayed an up to 200-fold higher catalytic efficiency than MAT. In addition, gene expression analysis, protein subcellular localization and heterologous expression in Nicotiana benthamiana were in agreement with the prominent role of TAT in acetylation of root-specific MIAs, thereby redefining the molecular determinants of the root MIA biosynthetic pathway. Finally, identification of TAT provided a convenient tool for metabolic engineering of MIAs in yeast enabling efficiently mixing different biosynthetic modules spatially separated in the whole plant. This combinatorial synthesis associating several enzymes from Catharanthus roseus resulted in the conversion of tabersonine in tailor-made MIAs bearing both leaf and root-type decorations. This article is protected by copyright. All rights reserved.
1365-313x Carqueijeiro, Ines Duge de Bernonville, Thomas Lanoue, Arnaud Dang, Thu-Thuy Teijaro, Christiana Paetz, Christian Billet, Kevin Mosquera, Angela Oudin, Audrey Besseau, Sebastien Papon, Nicolas Glevarec, Gaelle Atehortua, Lucia Clastre, Marc Giglioli-Guivarc'h, Nathalie Schneider, Bernd St-Pierre, Benoit Andrade, Rodrigo B O'Connor, Sarah E Courdavault, Vincent Journal Article England Plant J. 2018 May;94(3):469-484. doi: 10.1111/tpj.13868. Epub 2018 Mar 27.