Understanding and Exploiting the Biosynthesis of Diterpenoids in Euphorbia peplus

了解和利用大戟中二萜类化合物的生物合成

• 批准号: 506268802
• 负责人: Dr. Carsten Schotte
• 金额: --
• 依托单位: Abteilung Naturstoffbiosynthese
• 依托单位国家: 德国
• 项目类别: WBP Position
• 财政年份: 2022
• 资助国家: 德国
• 起止时间: 2021-12-31 至 2022-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/506268802?language=en
• 关键词: Understanding; Exploiting; Biosynthesis; Diterpenoids; Euphorbia

Terpenes represent one of the largest and most diverse class of specialized metabolites, including world health organisation (WHO)-listed essential drugs such as Taxol and Artemisinin. The polyoxygenated ingenol mebutate is a clinically important diterpenoid produced by the plant Euphorbia peplus. In 2012 it was approved by the Food and Drug Administration (FDA) for the treatment of actinic keratosis, a precancerous skin condition. At the moment it is under further evaluation for the treatment of basal cell carcinoma and the activation of latent HIV reservoirs in the human body. Despite the growing pharmaceutical importance of ingenol mebutate current supplies of this valuable metabolite are limited, as it is only produced in minute quanitites in planta and currently no feasible total synthesis has been reported. In this study we aim to apply a `multi-omics´ approach to elucidate the biosynthetic pathway towards ingenol mebutate in Euphorbia peplus. Genome and RNA sequencing data will be obtained and analysed to identify genes involved in the biosynthesis of ingenol mebutate and concomitantly produced diterpenoids. Candidate genes will be tested for activity in a robust Agrobacterium-mediated transient expression system in tobacco and via RNAi-experiments in hairy root tissue cultures of Euphorbia peplus. Once all enzymes are identified we aim to reconstitute the entire biosynthetic pathway in tobacco, thus providing the first total biosynthesis of ingenol mebutate in a heterologous host. The proposed project will shed light on the biosynthesis of a pharmaceutically important, understudied class of plant specialized metabolism and lay the foundation for future combinatorial and semi-synthetic synthesis of new-to-nature diterpenoids with fine-tuned biological activities.

萜烯是最大和最多样化的一类特殊代谢物，包括世界卫生组织（WHO）列出的基本药物，如紫杉醇和青蒿素。多氧巨大戟醇甲基丁酸酯是由植物大戟产生的临床上重要的二萜类化合物。2012年，它被美国食品和药物管理局（FDA）批准用于治疗光化性角化病，这是一种癌前皮肤病。目前，它正在进一步评估用于治疗基底细胞癌和激活人体内潜伏的HIV储库。尽管巨大戟醇甲基丁酸酯的药学重要性日益增加，但这种有价值的代谢物的当前供应是有限的，因为它仅在植物中以微量quanitites产生，并且目前还没有报道可行的全合成。在这项研究中，我们的目标是应用“多组学”的方法来阐明的生物合成途径巨大戟醇甲基丁酸在大戟。将获得并分析基因组和RNA测序数据，以鉴定参与巨大戟醇甲基丁酸酯生物合成的基因和伴随产生的二萜类化合物。候选基因将在烟草中稳健的农杆菌介导的瞬时表达系统中以及通过在大戟毛状根组织培养物中的RNAi实验来测试活性。一旦所有的酶被确定，我们的目标是重建烟草中的整个生物合成途径，从而提供了第一个总的巨大戟醇甲基丁酸酯在异源宿主的生物合成。拟议的项目将阐明生物合成的药学上重要的，未充分研究的一类植物专门的代谢，并奠定了基础，为未来的组合和半合成合成的新的天然二萜类化合物与微调的生物活性。