(725b) Discovering a Tomato Natural Product Biosynthetic Pathway Using an Integrated Synthetic Biology Approach

Plant natural products (PNPs) are chemicals of great pharmaceutical and nutritional significance due to their structural complexity and chemical diversity. PNP discovery traditionally relies on the extraction and isolation of metabolites from plants, facing limitations in cost and efficiency. Recent advances in plant genome sequencing provide an opportunity to accelerate the PNP discovery process by the identification and validation of plant biosynthetic pathways producing corresponding PNPs. However, systematic prediction and reconstitution of plant pathways remain challenging, mainly due to the size and complexity of plant genomes, and the difficulty of efficient in planta gene modification. Recent plant genomics studies reveal a large number of putative plant biosynthetic pathways clustered in plant genomes, which highlights the potential of a genomics-driven plant biosynthetic pathway prediction pipeline. Recent advances in synthetic biology highlight the opportunity to exploit heterologous hosts to elucidate PNP biosynthetic pathways and to efficiently produce novel PNPs, thereby bypassing many challenges associated with plant engineering. Thus, the interface of genomics, bioinformatics, and synthetic biology promises to advance PNP discovery, which will further our understanding of plant metabolism and complement current PNP manufacturing pipelines. Here, we report an innovative approach enabling rapid, high­throughput discovery of PNPs through predicted gene cluster reconstruction and metabolite analysis in heterologous hosts such as Saccharomyces cerevisiae. We demonstrated this approach for the discovery of a novel tomato pathway and corresponding phenolic compounds. We also characterized a key enzyme in the pathway that exhibits previously unknown bifunctional activity, which may lead to new biosynthetic mechanisms and products. The integration of a bioinformatics-driven plant biosynthetic pathway prediction pipeline and a yeast-based synthetic biology platform provides a promising platform for efficient novel PNP discovery and scalable biomanufacturing, which will ultimately advance the discovery and development of small-molecule drugs in yeast.