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: 10647767
• 负责人: Angad Mehta
• 金额: $ 29.51万
• 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/10647767
• 关键词: Acids; Algae; Amorpha; Anacystisnidulans; Antibiotics; Antineoplastic Agents; Area; Artemisinins; Bacteria; Biopolymers; Biotechnology; Carbon; Carbon Dioxide; Cells; 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; Life; Light; Metabolic; Mitochondria; Modeling; Molecular; Natural Products; Organelles; Organism; Paclitaxel; Pharmaceutical Preparations; Pharmacologic Substance; Photosynthesis; Plants; Saccharomyces cerevisiae; Saccharomycetales; Signal Transduction; Structure; System; Terpenes; Variant; Yeast Model System; Yeasts; chemical synthesis; cost; diene; drug synthesis; endosymbiont; improved; industrial production; metabolic engineering; mutant; natural product inspired; novel; success; synthetic biology; taxadiene; technology platform; 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 很大一部分重要的药物和医药中间体或者来源于植物， 由石油化工产品合成。几种对人类有很高价值的天然产物 寿命（例如，抗生素，药物如青蒿素，抗癌剂如紫杉醇（紫杉醇）等） 是由不适合工业生产的生物体产生的。虽然其中一些天然的 产品可以化学合成，其中几种化合物的复杂结构使化学 合成困难且商业上不可行。因此，越来越需要发展可持续和 易于处理的技术平台来合成这些药物和药物中间体。的 该提案的中心目标是开发一个可持续的技术平台， 和生物催化来合成与人类生命有重大关系的分子。我们设想这样做， 在酵母细胞内建立蓝细菌内共生体， 为酵母细胞提供ATP和同化碳（光合作用产生的），酵母细胞利用它来生产 重要的生物天然产物。这个平台将使我们能够将卓越的生物合成和 酵母的生物催化潜力，以蓝藻的光合能力，以发展一个可持续的和简单的 生物生产平台，生产对人类生命具有重要价值的天然产品（见下文）。（i） 酿酒酵母最近已被利用来生产高滴度的生物学重要的 分子如紫穗槐二烯和青蒿酸（青蒿素的前体）和紫杉二烯（一种关键的 紫杉醇的前体），（ii）我们以前已经开发了S.酿酒酵母/E.杆菌以 研究线粒体进化目前是工程酵母/蓝细菌内共生（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，我们将集中在三个关键领域：（i）我们设计了实验平台， 在模式蓝藻细长聚球藻和模式芽殖酵母之间建立内共生， S.啤酒。我们将通过工程设计新的蓝藻突变体来扩展这个平台， 内共生体我们将广泛表征工程酵母/蓝藻内共生，以开发 提高其稳定性、增长率和同质性的战略。(ii)我们将创造一个代谢物驱动的合成 通讯系统来控制内共生和优化我们的代谢工程平台。(iii)我们 将利用我们的光合内共生平台生产FDA批准的化合物的关键前体， 青蒿素和紫杉醇。这些研究将是我们实现发展一个 光合作用和遗传上易处理的内共生平台，用于生物生产生物学上重要的 新型天然产物以及生物聚合物和生物燃料等分子。