Harnessing light and model endosymbiosis to produce natural products

利用光和模型内共生来生产天然产品

• 批准号: 10361643
• 负责人: Angad Mehta
• 金额: $ 28.57万
• 依托单位: UNIVERSITY OF ILLINOIS AT URBANA-CHAMPAIGN
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-20 至 2026-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10361643
• 关键词: Acids; Amorpha; Anacystisnidulans; Antibiotics; Antineoplastic Agents; Area; Artemisinins; Bacteria; Biological; Biopolymers; Biotechnology; Carbon; Carbon Dioxide; Cells; Chemicals; Chimera organism; Chloroplasts; Communication; Complex; Couples; Coupling; Cyanobacterium; Data; Development; Engineering; Escherichia coli; Ethanol; Eukaryotic Cell; Evolution; FDA approved; Fermentation; Gene Expression; Genetic; Genetic Engineering; Glucose; Glycerol; Goals; Growth; Health; Human; Industrialization; Life; Light; Metabolic; Mitochondria; Modeling; Molecular; Natural Products; Organelles; Organism; Paclitaxel; Pharmaceutical Preparations; Pharmacologic Substance; Photosynthesis; Plants; Production; Saccharomyces cerevisiae; Saccharomycetales; Signal Transduction; Structure; System; Terpenes; Variant; Yeast Model System; Yeasts; base; chemical synthesis; cost; diene; endosymbiont; improved; metabolic engineering; mutant; novel; success; synthetic biology; taxadiene; tool

Project Abstract A large portion of important drugs and pharmaceutical intermediates either originate from plants or are synthesized from petrochemical-based products. Several natural products of significantly high value to human life (e.g., antibiotics, pharmaceuticals like Artemisinin, anticancer agents like Taxol (paclitaxel) among others) have been produced by organisms that are not optimal for industrial production. Although some of these natural products can be chemically synthesized, the complex structures of several of these compounds makes chemical synthesis difficult and commercially infeasible. As a result, there is increasing need to develop sustainable and readily tractable technological platforms to synthesize these drugs and pharmaceutical intermediates. The central objective of this proposal is to develop a sustainable technological platform that harnesses light energy and biocatalysis to synthesize molecules of significant relevance to human life. We envision doing this by establishing cyanobacterial endosymbionts within yeasts cells, such that the endosymbiotic cyanobacteria provide ATP and assimilated carbon (generated from photosynthesis) to the yeast cells, which utilize it to produce biologically important natural products. This platform will allow us to couple the remarkable biosynthetic and biocatalytic potential of yeast to the photosynthetic ability of cyanobacteria to develop a sustainable and simple bioproduction platform to produce natural products of significant value to human life (vide infra). Premise: (i) Saccharomyces cerevisiae has been recently harnessed to produce high titers of biologically important molecules such as amorphadiene and artemisinic acid, (precursors to Artemisinin) and taxadiene (a key precursor to Taxol), (ii) we had previously developed model endosymbiosis between S. cerevisiae /E. coli to study mitochondrial evolution are currently engineering yeast/cyanobacteria endosymbiosis (preliminary data in Specific Aim 1) and (iii) our preliminary data on engineering model yeast/cyanobacteria endosymbiosis. In this proposal, we will focus on three key areas: (i) We have engineered experimental platform to establish endosymbiosis between model cyanobacteria, Synechococcus elongatus, and model budding yeast, S. cerevisiae. We will expand this platform by engineering novel cyanobacterial mutants as putative endosymbionts. We will extensively characterize the engineered yeast/cyanobacteria endosymbiosis to develop strategies to improve their stability, growth rate and homogeneity. (ii) We will create a metabolite-driven synthetic communication system to control endosymbiosis and optimize our platform for metabolic engineering. (iii) We will utilize our photosynthetic endosymbiotic platform to produce key precursors of FDA approved compounds, Artemisinin and Taxol. These studies will be the first step towards our long-term goal of developing a photosynthetic and genetically tractable endosymbiotic platform for the bioproduction of biologically important novel natural products as well as molecules like biopolymers and biofuels.

项目摘要 很大一部分重要药物和医药中间体要么来自植物，要么 以石化为基础的产品合成。几种对人类具有显著高价值的天然产品 生命(如抗生素、青蒿素等药物、紫杉醇等抗癌药物等) 都是由不适合工业生产的有机体产生的。尽管其中一些是自然的 产品可以化学合成，其中几种化合物的复杂结构使化学 合成困难，商业上不可行。因此，越来越需要发展可持续和 合成这些药物和医药中间体的易于操作的技术平台。这个 这项提议的中心目标是开发一个可持续的技术平台，利用光能 和生物催化来合成与人类生活有重大关系的分子。我们设想通过以下方式做到这一点 在酵母细胞内建立蓝藻内共生菌，以使内共生蓝藻 向酵母细胞提供三磷酸腺苷和同化碳(通过光合作用产生)，酵母细胞利用它生产 具有重要生物学意义的天然产物。这个平台将允许我们将非凡的生物合成和 酵母生物催化潜力对蓝藻光合作用能力的可持续发展 生物生产平台，生产对人类生活有重大价值的天然产品(见下文)。前提：(I) 酿酒酵母最近被利用来生产具有重要生物学意义的高滴度 阿莫菲二烯和青蒿酸(青蒿素的前体)和紫杉二烯(一种关键)等分子 紫杉醇的前体)，(Ii)我们以前已经在酿酒酵母/大肠杆菌之间建立了模型内共生，以 研究线粒体进化目前正在改造酵母/蓝藻内共生(初步数据在 具体目标1)和(Iii)我们的工程模型酵母/蓝藻内共生的初步数据。 在这项建议中，我们将重点关注三个关键领域：(I)我们已经设计了实验平台，以 在模型蓝藻、长聚球藻和模型发芽酵母之间建立内共生， 酿酒酵母。我们将通过设计新的蓝藻突变体来扩展这一平台 内生共生菌。我们将广泛研究工程酵母/蓝藻内共生的特性，以开发 提高其稳定性、增长率和同质性的战略。(Ii)我们将创造一种代谢物驱动的合成 控制内共生和优化代谢工程平台的通信系统。(Iii)我们 将利用我们的光合作用内共生平台生产FDA批准的化合物的关键前体， 青蒿素和紫杉醇。这些研究将是迈向我们长期目标的第一步，即制定一项 光合作用和遗传上易驯化的内共生平台，用于生物生产具有重要生物学意义 新的天然产品以及生物聚合物和生物燃料等分子。