CAREER: Enhancing probiotic yeast colonization for stable in situ biomanufacturing

治疗肠道疾病是困难的，因为身体在达到目标之前会抵制许多治疗。 益生菌酵母可以为肠道产生许多治疗药物，但这种酵母不会在体内存活很长时间。为了解决这个问题，这个教师早期职业发展计划（CAREER）项目将探索提高基因工程益生菌驻留在肠道中的能力的策略。酵母菌将被设计成附着在肠道内粘糊糊的、富含糖的粘液上并吃掉它们。将开发新的课程，培养学生的微生物工程技能。高中生将有机会进化益生菌以实现更快的生长，并在这样做的同时学习进化和遗传学。本科生将学习如何通过与NCSU生物技术计划（BIT）合作开发的新实验室课程生物展望新基因。总之，这项工作将提高向肠道输送药物的能力，并将加强北卡罗来纳州代表性不足的高中生和本科生的教育。布拉氏菌（Sb）是一种广泛可用的益生菌，通常被认为是安全的。它的独特之处在于它是唯一的益生菌酵母（甚至是真核生物），因此为原位生物制造带来了一系列独特的优势。它的高分泌速率和传感能力已被用于产生有前途的活治疗剂。然而，先前的报告表明Sb不会在人类或大鼠肠道中定植超过一周。肠道微生物有两种非排他性的策略来殖民肠道。一种是粘附在肠壁上，第二种策略是利用肠道营养。 这项工作将探讨两种策略，提高锑殖民，测试的假设，增加锑的粘液粘附，以及其代谢粘液聚糖的能力，将改善其停留时间。 除了这些研究目标之外，在肠道中观察到的Sb和其他益生菌的适应性进化的强烈特征也激发了当地高中外展模块的开发，该奖项反映了NSF的法定使命，并被认为是值得通过使用基金会的智力价值和更广泛的影响审查标准。

Treating gut disease is difficult because the body digests many therapies before they reach their target. Probiotic yeast can produce many therapeutics for the gut, but this yeast does not live for a long time in the body. To solve this problem, this Faculty Early Career Development Program (CAREER) project will explore strategies to improve the ability of genetically engineered probiotics to reside in the gut. Yeast will be engineered to attach to and eat the slimy, sugar-rich mucus that line the intestines. New courses will be developed to train students in skills for microbial engineering. High school students will have the opportunity to evolve probiotics for faster growth and learn about evolution and genetics while doing so. Undergraduates will learn how to bioprospect for new genes via a new lab course developed in partnership with the NCSU Biotechnology Program (BIT). Taken together, this work will advance the ability to deliver medicines to the gut and will enhance the education of underrepresented high schoolers and undergraduates in North Carolina.S. boulardii (Sb) is a widely available probiotic with generally-regarded-as-safe status. It is unique in that it is the only probiotic yeast (or even eukaryote), and so brings with it a unique set of advantages for in situ biomanufacturing. Both its high secretion rate and sensing ability have been used to generate promising live therapeutics. However, prior reports have indicated that Sb does not colonize the human or rat intestine for longer than one week. There are two nonexclusive strategies that commensal microbes use to colonize the gut. One is to adhere to intestinal walls, and a second strategy is to utilize gut nutrients. This work will explore both strategies for enhancement of Sb colonization, testing the hypothesis that increasing Sb’s adherence to mucus, as well as its ability to metabolize mucus glycans, would improve its residence time. Beyond these research objectives, the strong signatures of adaptive evolution observed in the gut for Sb and other probiotics have also inspired the development of an outreach module for local high schools, as well as an undergraduate laboratory course harnessing horizontal gene transfer for biotech applications.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.