CAREER: Enhancing probiotic yeast colonization for stable in situ biomanufacturing

肠道疾病的治疗是困难的，因为身体在达到目标之前消化了许多疗法。益生菌酵母可以产生许多肠道治疗药物，但这种酵母在体内不会存活很长时间。为了解决这个问题，这个学院早期职业发展计划(Career)项目将探索提高基因工程益生菌在肠道中生存的能力的策略。酵母将被改造成附着并吃掉粘稠的、富含糖的粘液，这些粘液排列在肠道里。将开发新的课程，培训学生微生物工程技能。高中生将有机会进化益生菌以实现更快的生长，并在进化和遗传学的同时学习。本科生将通过与NCSU生物技术计划(BIT)合作开发的一门新的实验室课程，学习如何对新基因进行生物勘探。总而言之，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未被充分代表的高中生和本科生的教育。布氏葡萄球菌(SB)是一种广泛获得的益生菌，具有普遍被认为是安全的状态。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经被用来产生有希望的活疗法。然而，先前的报告表明，SB在人或大鼠肠道中的定植时间不超过一周。共生微生物有两种非排他性的策略来定植肠道。一种是附着在肠壁上，第二种策略是利用肠道营养。这项工作将探索两种增强Sb定植的策略，测试增加Sb对粘液的粘附性以及其代谢粘液多糖的能力将改善其停留时间的假设。除了这些研究目标之外，在SB和其他益生菌的肠道中观察到的适应进化的强烈信号也激励着为当地高中开发一个扩展模块，以及一个利用水平基因转移用于生物技术应用的本科实验室课程。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。

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.