Articles
Enhancing artemisinin biosynthesis in A. annua through transient expression of five key pathway genes
Article number
1462_3
Pages
15 – 24
Language
English
Abstract
Since its isolation in the early 1970s artemisinin marked a breakthrough in malaria treatment around the world, helping to save millions of lives.
Nowadays, artemisinin-based combination therapies (ACT) are recommended by the World Health Organization (WHO) as the primary treatment for malaria.
Despite extensive efforts to enhance production through synthetic and semi-synthetic approaches, extraction from Artemisia annua plants remains the main source of artemisinin, and its global availability continues to be a challenge for ensuring a stable and affordable supply of antimalarial drugs.
In this work, we established a transient expression protocol in A. annua plants grown under in vitro conditions, using GFP as a reporter gene.
This approach was then applied to assess the effects of overexpressing five key genes involved in artemisinin biosynthetic pathway: Farnesyl pyrophosphate synthase (FPS), Amorpha-4-11-diene synthase (ADS), Cytochrome P450 monooxygenase (CYP71AV1), Artemisinic aldehyde reductase (DBR2) and Aldehyde dehydrogenase (ALDH1). To this end, we engineered a multigene vector harboring all these five genes, placed under the transcriptional control of CaMV 35S constitutive promoter.
The effects of the expression of this multigene vector were evaluated through LC-MS analysis, to quantify artemisinin and its metabolic intermediate.
This study could help to sheds light on potential regulatory bottlenecks in the artemisinin pathway and provides insights for developing metabolic engineering strategies aimed at increasing artemisinin content in A. annua plants.
Nowadays, artemisinin-based combination therapies (ACT) are recommended by the World Health Organization (WHO) as the primary treatment for malaria.
Despite extensive efforts to enhance production through synthetic and semi-synthetic approaches, extraction from Artemisia annua plants remains the main source of artemisinin, and its global availability continues to be a challenge for ensuring a stable and affordable supply of antimalarial drugs.
In this work, we established a transient expression protocol in A. annua plants grown under in vitro conditions, using GFP as a reporter gene.
This approach was then applied to assess the effects of overexpressing five key genes involved in artemisinin biosynthetic pathway: Farnesyl pyrophosphate synthase (FPS), Amorpha-4-11-diene synthase (ADS), Cytochrome P450 monooxygenase (CYP71AV1), Artemisinic aldehyde reductase (DBR2) and Aldehyde dehydrogenase (ALDH1). To this end, we engineered a multigene vector harboring all these five genes, placed under the transcriptional control of CaMV 35S constitutive promoter.
The effects of the expression of this multigene vector were evaluated through LC-MS analysis, to quantify artemisinin and its metabolic intermediate.
This study could help to sheds light on potential regulatory bottlenecks in the artemisinin pathway and provides insights for developing metabolic engineering strategies aimed at increasing artemisinin content in A. annua plants.
Publication
Authors
F. Pietrolucci, S. Negri, V. Valesi, M. Commisso, F. Guzzo, L. Avesani
Keywords
artemisinin, A. annua, transient expression, metabolic engineering, LC-MS
Online Articles (28)