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，以治疗具有挑战性的疾病。该项目的目标是利用个体自身的微生物群在肠道中制造治疗性分子。研究人员的合作团队，跨越基因工程和人类细胞生物学的专业知识，将研究如何设计益生菌酵母对肠道环境的反应，并利用这些信息设计酵母菌株，感知它们在肠道中并产生维生素a。这项工作的发现可以为各种微生物的其他“即时”生物制造方法提供信息。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“向上跳跃”和“科学家一天”项目，从STEM领域代表性不足的背景中招募学生。这个跨学科研究团队的目标是制定策略，使博氏酵母（一种非致病性益生菌酵母）能够根据肠道状况产生高滴度的生物分子。在肠道样条件下，益生菌酵母增强蛋白质分泌的基因敲除组合将被确定。将寻求和测试多种策略，以通过表达专用输出蛋白和程序化自裂解来增强生物分子的递送。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宏基因组测序进行评估。这项工作通过建立创新策略，使一般生物分子释放到哺乳动物肠道，扩大了生物制造的前沿。此外，这将是第一次在肠道中设计小分子分泌的努力，也是第一次在酵母中设计自我裂解的努力。该奖项由eng - cbet -细胞与生物化学工程项目和bio - mcb -系统与合成生物学集群共同资助。该奖项反映了美国国家科学基金会的法定使命，并通过使用基金会的知识价值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.