Engineering probiotic yeast to release intracellular molecules into the mammalian gut

在体内制造所需的分子提供了一种直接提供治疗方法的方法，以应对具有挑战性的治疗条件。这个项目的目标是利用个人自己的微生物群在肠道中制造治疗分子。这个合作的研究团队跨越了基因工程和人类细胞生物学的专业知识，他们将研究如何设计益生菌酵母来对肠道环境做出反应，并利用这些信息来设计能感觉到它们在肠道内并产生维生素A的酵母菌株。这项工作的发现可以为其他各种微生物的“及时”生物制造方法提供借鉴。该项目将包括本科生研究人员，并通过北卡罗来纳州立大学的三人向上绑定和科学家一天计划引入STEM中代表性不足的背景的学生。这个跨学科的研究团队旨在开发能够在布拉酵母菌(SB)中高效生产生物分子的策略，这种酵母是一种非致病的益生菌酵母，可以响应肠道条件。在类似肠道的条件下，益生菌酵母能够促进蛋白质分泌的基因敲除组合将被识别出来。将寻求和测试多种策略，通过表达专用输出蛋白和程序化自我裂解来增强生物分子的传递。两种模型化合物β-胡萝卜素和抗菌肽的释放将分别使用定量蛋白质组学和超基因组测序进行评估。这项工作通过建立创新的策略，使一般生物分子能够释放到哺乳动物的肠道中，从而扩大了生物制造的前沿。此外，这将是第一个在肠道中设计小分子分泌的努力，也是第一个在酵母中设计自我裂解的努力。该奖项由ENG-CBET-细胞和生化工程计划以及BIO-MCB-系统和合成生物学集群联合资助。该奖项反映了NSF的法定使命，并通过使用基金会的智力优势和更广泛的影响审查标准进行评估，被认为值得支持。

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.