(711e) Harnessing Synthetic Biology to Uncover & Engineer Plant Natural Product Biosynthesis in Yeast

Plant natural products are valuable compounds with diverse therapeutic activities whose biosynthesis is poorly understood due to the challenges in functionally identifying and characterizing pathway enzymes. While the recent discovery of plant gene clusters offers the potential to accelerate pathway discovery in plants, current enzyme characterization strategies limit the efficiency and accuracy of subsequent pathway elucidation. We describe an alternative approach, fueled by recent methodological advances in synthetic biology, for the rapid elucidation of complex plant biosynthetic pathways through the step-wise reconstruction of plant gene clusters in Saccharomyces cerevisiae. We demonstrate this approach for the full elucidation of one of the most complex gene clusters identified to date. Our work not only offers novel insight into several of the enzymatic steps in the pathway, but also provides yeast strains engineered to produce previously inaccessible pathway intermediates and novel derivatives, thereby providing platforms for advancing drug discovery. The synthetic biology-based approach offers several advantages over traditional elucidation strategies based on gene silencing in the plant host and in vitro enzyme characterization, including characterization of pathway metabolites in a clean host background, direct supply of pathway intermediates for structural validation, and simultaneous development of scalable synthesis platforms for intermediates and novel derivatives.