Transient production of artemisinin in Nicotiana benthamiana is boosted by a specific lipid transfer protein from A. annua
| Autor: | Peter E. Brodelius, Arman Beyraghdar Kashkooli, Linda Olofsson, Harro J. Bouwmeester, Alexander R. van der Krol, Mathieu Pottier, Adrienne Sallets, Bo Wang, Norbert C.A. de Ruijter, Marc Boutry, Hieng-Ming Ting |
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| Rok vydání: | 2016 |
| Předmět: |
0106 biological sciences
0301 basic medicine Artemisia annua Nicotiana benthamiana Heterologous Bioengineering Biology 01 natural sciences Applied Microbiology and Biotechnology Metabolic engineering 03 medical and health sciences chemistry.chemical_compound Biosynthesis Tobacco Laboratorium voor Plantenfysiologie Pleiotropic Drug Resistance protein Plant Proteins fungi food and beverages Laboratorium voor Celbiologie biology.organism_classification Artemisinins Apoplast Biosynthetic Pathways Laboratory of Cell Biology Genetic Enhancement 030104 developmental biology ABC transporters Metabolic Engineering chemistry Biochemistry Artemisinin Lipid transfer proteins Heterologous expression EPS Carrier Proteins Plant lipid transfer proteins Laboratory of Plant Physiology Metabolic Networks and Pathways 010606 plant biology & botany Biotechnology |
| Zdroj: | Metabolic Engineering, 38, 159-169 Metabolic Engineering 38 (2016) Metabolic Engineering |
| ISSN: | 1096-7176 |
| DOI: | 10.1016/j.ymben.2016.07.004 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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