Engineering probiotic yeast to release intracellular molecules into the mammalian gut

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Manufacturing desired molecules in the body offers a way to directly deliver therapies for challenging to treat conditions. The goal of this project is to harness an individual’s own microbiome to manufacture therapeutic molecules in the gut. The collaborative team of researchers, spanning expertise in genetic engineering and human cell biology, will investigate how to engineer probiotic yeast to react to the gut environment and use this information to design yeast strains that sense they are in the gut and produce vitamin A. The findings from this work can inform other "just-in-time" biomanufacturing approaches in a variety of microbes. This project will include undergraduate researchers and bring in students from under-represented backgrounds in STEM via the TRIO Upward Bound and Scientist for a Day programs at North Carolina State University. The interdisciplinary research team aims to develop strategies that enable high-titer production of biomolecules in Saccharomyces boulardii (Sb), a non-pathogenic probiotic yeast, in response to gut conditions. Combinations of gene knockouts that enhance protein secretion under gut-like conditions by probiotic yeast will be identified. Multiple strategies will be pursued and tested to enhance biomolecule delivery through expression of dedicated export proteins and programmed self-lysis. The release of two model compounds, β-carotene and antimicrobial peptides, will be evaluated using quantitative proteomics and metagenome sequencing, respectively. This work expands the frontiers of biomanufacturing by establishing innovative strategies enabling general biomolecule release into the mammalian gut. Furthermore, this will be the first effort to engineer small molecule secretion in the gut and the first effort to engineer self-lysis in yeast.This award is jointly funded by the ENG-CBET-Cellular and Biochemical Engineering program and BIO-MCB-Systems and Synthetic Biology cluster.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.