CAREER: Synthetic Biology to Understand and Harness Plant Enzyme Complexes for Natural Product Synthesis in Yeast

从药用植物中获取具有生物活性的天然产物在药物生产中很重要。然而，以农业为基础的供应链容易受到环境变化和全球危机的影响，因为许多药用植物只能在气候和冲突脆弱地区种植。微生物生物制造是生产具有生物活性的植物天然产物的一种强有力的替代方法。工程微生物可以在短时间内通过密闭容器发酵产生具有生物活性的植物天然产物，从而为加强供应链提供了一种有效的方法。这个项目使用新兴的合成生物学方法来设计微生物。特别是，该项目在酵母中重建了源自植物的生物合成机器，以生产有价值的植物天然产品。这项研究为女学生开展了协同的教育和研究活动，最终培养了合成生物学领域的下一代女性领导人。本项目面向高中学生和植物生物学或工程专业的本科生，以及成熟的工程师。这些活动包括暑期项目、跨学科教育和研究培训，以及将工程师与领先的合成生物学初创公司和世界知名女企业家联系起来的机会。合成生物学的发展为在微生物宿主（如面包酵母）中通过重建植物来源的异源生物合成途径来生产植物天然产物打开了一扇大门。植物中许多关键的调控活动和机制仍然知之甚少，这极大地限制了植物天然产物的生物制造。植物可以通过形成动态的植物酶复合物来调节其天然产物的生物合成途径。这些复合物广泛存在于植物中，被认为是快速动态调节植物天然产物合成的关键空间组织机制。该项目通过基于合成生物学的方法来重建和表征酵母中的植物复合物，以推进植物天然产物的生物合成，从而解决了这一知识缺口。选择药用植物玫瑰花（Catharanthus roseus）为例进行研究，因为玫瑰花所产生的有价值的单萜吲哚类生物碱以及其途径和复合物已被部分表征。本项目开发的合成生物学工具和方法使植物酶在酵母中的动态组装能够进行复杂的重建和表征，提高对植物翻译后调控机制的理解，最终促进有价值的植物天然产物的微生物生物制造。该奖项反映了美国国家科学基金会的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。

Sourcing bioactive natural products from medicinal plants is important in drug production. However, the agriculture-based supply chain is susceptible to environmental changes as well as global crises, as many medicinal plants can only be grown in climate and conflict vulnerable regions. Microbial biomanufacturing is a powerful alternative approach to produce bioactive plant natural products. Engineered microorganisms can produce bioactive plant natural products in a short period of time by fermentation in closed vessels, thus providing an efficient approach to strengthen the supply chain. This project uses emerging synthetic biology methods to engineer microorganisms. In particular, the project reconstitutes, in yeast, the biosynthetic machinery derived from plants to produce valuable plant natural products. This study develops synergistic educational and research activities for women students, ultimately fostering the next generation of women leaders in the field of synthetic biology. The project targets high school students and undergraduate students majoring in plant biology or engineering, as well as established engineers. The activities include summer programs, interdisciplinary education, and research training, as well as opportunities for connecting engineers with leading synthetic biology startups and world-famous women entrepreneurs.The development of synthetic biology has opened the gate to a powerful alternative approach for production of plant natural products in a microbial host, such as baker’s yeast, by reconstructing the plant-derived heterologous biosynthetic pathways. Many critical regulatory activities and the machinery in plants remain poorly understood, which significantly limits plant natural product biomanufacturing. Plants can regulate their natural product biosynthetic pathways by forming dynamic plant enzyme complexes. These complexes widely exist in plants and are believed to be critical spatial organization machinery that promptly and dynamically regulates plant natural product synthesis. This project addresses this knowledge gap through a synthetic biology-based approach to rebuilding and characterizing plant complexes in yeast to advance plant natural product biosynthesis. The medicinal plant Catharanthus roseus is chosen as the example to study due to the valuable pharmaceutical monoterpene indole alkaloids it produces and the pathways and complexes that have been partially characterized. Synthetic biology tools and methods developed in this project enable the dynamic assembly of plant enzymes in yeast for complex reconstruction and characterization, improve the understanding of the post-translational regulation mechanism in plants, and ultimately advance microbial biomanufacturing of valuable plant natural products.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.