CAREER: Enhancing probiotic yeast colonization for stable in situ biomanufacturing

治疗肠道疾病是很困难的，因为身体在许多疗法到达目标之前就消化了。益生菌酵母菌可以产生许多治疗肠道的药物，但这种酵母菌不能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在肠道中的生存能力的策略。酵母菌将被改造成附着并吃掉黏稠的、富含糖的黏液。将开设新课程，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加快生长，并在此过程中学习进化和遗传学。本科生将通过与NCSU生物技术计划（BIT）合作开发的新实验课程学习如何对新基因进行生物勘探。总的来说，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。博氏菌（Sb）是一种广泛使用的益生菌，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（甚至真核生物），因此它为原位生物制造带来了一系列独特的优势。它的高分泌率和传感能力已被用于产生有前途的活体治疗。然而，先前的报道表明Sb在人或大鼠肠道的定殖时间不超过一周。共生微生物利用两种非排他性策略在肠道中定居。一个是坚持肠壁，第二个策略是利用肠道营养物质。本研究将探索两种增强Sb定植的策略，验证增加Sb对黏液的粘附以及代谢黏液聚糖的能力将延长其停留时间的假设。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推广模块的开发，以及利用生物技术应用水平基因转移的本科实验课程。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。

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.