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 大部分重要药物和药物中间体源自植物，或者是 由基于石化的产品合成。几种对人类价值显着高的天然产品 生命（例如，抗生素，诸如Artemisinin，例如紫杉醇（紫杉醇）等抗癌药等药物） 是由不适合工业生产最佳的生物生产的。尽管其中一些很自然 产品可以化学合成，其中几种化合物的复杂结构使化学 综合困难且商业上不可行。结果，越来越需要发展可持续性和 易于处理的技术平台，可以合成这些药物和药物中间体。这 该建议的核心目标是开发一个可持续的技术平台，以利用轻能 和生物催化，以合成与人类生命有重大相关性的分子。我们设想这样做 在酵母菌细胞中建立蓝细菌内共生菌，使得内共生蓝细菌 提供ATP和同化碳（从光合作用）到酵母细胞，并利用它产生 生物学上重要的天然产品。这个平台将使我们能够搭配出色的生物合成和 酵母对蓝细菌的光合作用能力的生物催化潜力发展可持续和简单 生物生产平台可生产对人类生命具有重要价值的天然产物（视频下文）。前提：（i） 酿酒酵母的糖疗法最近已被利用，生成了较高的生物学重要性滴度 分子，例如非甲二烯和artemisinic酸（青蒿素的前体）和纳税二烯（关键 紫杉醇的前体），（ii）我们以前在酿酒酵母 /e之间开发了模型内共生。大肠杆菌 研究线粒体进化目前是工程酵母/蓝细菌内共生症（初步数据 具体目的1）和（iii）我们有关工程模型酵母/蓝细菌内共生的初步数据。 在此建议中，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 在模型蓝细菌，链球菌和模型萌芽的酵母菌之间建立内共生 S. cerevisiae。我们将通过工程新颖的蓝细菌突变体作为推定来扩展这个平台 内共生体。我们将广泛地表征工程酵母/蓝细菌内共生的发展以发展 提高其稳定性，增长率和同质性的策略。 （ii）我们将创建一个代谢物驱动的合成 沟通系统控制内生物关系并优化我们的代谢工程平台。 （iii）我们 是否会利用我们的光合性内共生平台来生成FDA认可化合物的关键前体， 青蒿素和紫杉醇。这些研究将是朝着我们建立一个长期目标的第一步 光合作用和遗传上可触犯的内共生式平台，用于生物生物生物生物生物生物生产 新型天然产物以及生物聚合物和生物燃料等分子。