Transient production of artemisinin in Nicotiana benthamiana is boosted by a specific lipid transfer protein from A. annua
Journal
Metabolic Engineering
Journal Volume
38
Pages
159
Date Issued
2016-11-01
Author(s)
Wang, Bo
Kashkooli, Arman Beyraghdar
Sallets, Adrienne
de Ruijter, Norbert C.A.
Olofsson, Linda
Brodelius, Peter
Pottier, Mathieu
Boutry, Marc
Bouwmeester, Harro
van der Krol, Alexander R.
Abstract
© 2016 International Metabolic Engineering Society Our lack of full understanding of transport and sequestration of the heterologous products currently limit metabolic engineering in plants for the production of high value terpenes. For instance, although all genes of the artemisinin/arteannuin B (AN/AB) biosynthesis pathway (AN-PW) from Artemisia annua have been identified, ectopic expression of these genes in Nicotiana benthamiana yielded mostly glycosylated pathway intermediates and only very little free (dihydro)artemisinic acid [(DH)AA]. Here we demonstrate that Lipid Transfer Protein 3 (AaLTP3) and the transporter Pleiotropic Drug Resistance 2 (AaPDR2) from A. annua enhance accumulation of (DH)AA in the apoplast of N. benthamiana leaves. Analysis of apoplast and cell content and apoplast exclusion assays show that AaLTP3 and AaPDR2 prevent reflux of (DH)AA from the apoplast back into the cells and enhances overall flux through the pathway. Moreover, AaLTP3 is stabilized in the presence of AN-PW activity and co-expression of AN-PW+AaLTP3+AaPDR2 genes yielded AN and AB in necrotic N. benthamiana leaves at 13 days post-agroinfiltration. This newly discovered function of LTPs opens up new possibilities for the engineering of biosynthesis pathways of high value terpenes in heterologous expression systems.
Subjects
ABC transporters | Artemisia annua | Artemisinin | Lipid transfer proteins | Nicotiana benthamiana | Pleiotropic Drug Resistance protein
Publisher
ACADEMIC PRESS INC ELSEVIER SCIENCE
Type
journal article