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

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.