Engineering Specialized Organelles in Yeast for Compartmentalized Production of Therapeutics

/Abstract Eukaryotic cells utilize organelles to compartmentalize biochemical processes, ensuring spatial control over complex metabolism. This project aims to re-engineer the peroxisome and vacuole organelles to be synthetic, specialized organelles for engineered metabolic pathways. Accordingly, this project has significance both in directly testing, and likely expanding, the current understanding of the biology of these organelles and in the engineering of platform strains that will enable the unicellular microbe Saccharomyces cerevisiae to mimic the tissue compartmentalization observed in many plants for the synthesis of many natural products used as therapeutics in the clinic. Specifically, the reprogramming of the following key organelle properties will be targeted: organelle biogenesis and morphology, membrane permeability to small molecules, rate and efficiency of soluble protein import, and membrane protein trafficking to gain selective small molecule import/export. These engineering efforts will establish design rules for these organelle functions as well as develop powerful tools for the metabolic engineering of complex, therapeutic molecules with the preferred fermentation host, Saccharomyces cerevisiae. Accordingly, this unicellular microbe can be a chassis capable of compartmentalizing parts of a pathway in the peroxisome, vacuole, and cytoplasm. In the proposed work, increased flux through the benzylisoquinoline alkaloid pathway and control over which products are biosynthesized will be achieved using the specialized organelles engineered in this work. Project Number: 1R35GM164064-01 | Fiscal Year: 2026 | NIH Institute/Center: National Institute of General Medical Sciences (NIGMS) | Principal Investigator: John Dueber | Institution: UNIVERSITY OF CALIFORNIA BERKELEY, BERKELEY, CA | Award Amount: $387,596 | Activity Code: R35 | Study Section: Special Emphasis Panel[ZRG1 MCST-Q (55)] View on NIH RePORTER: https://reporter.nih.gov/project-details/11331702