Transition: Metabolomics-driven understanding of rules that coordinate metabolic responses and adaptive evolution of synthetic biology chassis

这一过渡奖支持首席研究员接受现代代谢组学和工程生物学方法的培训，使他的实验室能够破译和控制微生物新陈代谢。微生物代谢分配碳和能源资源，以支持生物质生长和生物生产。在合成生物学应用中，异源生物合成的实施会扰乱细胞供应链，给微生物带来代谢负担。这一问题导致生物技术中使用的微生物种群发生不可预测的生理变化，对合成生物微生物菌株的商业化构成了障碍。到目前为止，对代谢负荷如何影响代谢物产生、酶反应速率和细胞适应的了解仍然很少。随着这个问题的焦点，PI正在学习和应用新技术，以揭示代谢负担的潜在原因、程度和影响，这些负担导致生物技术中使用的微生物细胞的代谢或进化变化。该项目提供了关于压力条件下微生物生命规则的新知识，并为建立高性能的生物制造主力铺平了道路。此外，该项目还培训研究生、本科生和高中生进行跨学科的生物技术研究。首席研究员正在与林肯大学合作，组织夏季研究和劳动力发展合作。林肯大学是一所历史上的黑人学院和大学。这一奖项使首席研究员能够将他的研究重点转移到一个新的领域，即综合代谢组学、工程生物学和生物制造。所追求的职业发展有两个目标，第一个目标是学习现代液质联用(LC-MS)技术，这些技术可以用于进行同位素辅助代谢组学，以发现新的代谢物，描绘功能途径，确定酶反应热力学，并破译基因-表型关系。第二个目标是获得基于CRISPR的基因编辑工具，这些工具可用于重新编程调节体系结构和代谢途径。这项研究采用了新技术，并结合碳13代谢通量分析，分析了一种在不同生物反应器条件下用于天然产物生物合成的工业酵母底盘(酿酒酵母)。所进行的系统分析提高了对酵母菌细胞在代谢压力下的酶反应、代谢产物抑制和适应性细胞反应的驱动力的理解。新的见解和原则有望促进有效生物生产的微生物途径的设计和控制。这一奖项反映了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。

This Transitions award supports the principal investigator to be trained in modern metabolomics and engineering biology methods, enabling his lab to decipher and control microbial metabolism. Microbial metabolism distributes carbon and energy resources to support biomass growth and bio-productions. During synthetic biology applications, the implementation of heterologous biosynthesis can disrupt the cellular supply chains and impose metabolic burdens on microorganisms. This issue causes microbial populations used in biotechnology, to undergo unpredictable physiological changes, posing a roadblock for commercializing synthetic biology microbial strains. Until now, the understanding of how metabolic burdens affect metabolite productions, enzyme reaction rates, and cellular adaptations is still poor. With this problem in focus, the PI is learning and applying new technologies to uncover the underlying causes, extent, and effects of metabolic burdens that lead to metabolic or evolutionary changes in microbial cells that are used in biotechnology. This project delivers new knowledge on the rules of microbial life under stressed conditions and paves the way for building high performance biomanufacturing workhorses. Moreover, the project trains graduate students, undergraduates, and high school students in interdisciplinary biotechnology research. The principal investigator is partnering with Lincoln University, a Historically Black College and University, to organize both summer research and workforce development collaborations. This award enables the principal investigator to transition the focus of his research to a new area at the interface of integrative metabolomics, engineering biology, and biomanufacturing. The professional development that is pursued has two aims, the first of which is learning modern liquid chromatography–mass spectrometry (LC-MS) techniques that can be used to perform isotope assisted metabolomics to discover new metabolites, delineate functional pathways, determine enzyme reaction thermodynamics, and decipher genotype-phenotype relations. The second aim is to acquire CRISPR-based gene editing tools that can be used to re-program regulatory architecture and metabolic pathways. The research undertaken employs the new techniques, along with carbon-13 metabolic flux analysis, to analyze a model industrial yeast chassis (Saccharomyces cerevisiae) for the biosynthesis of natural products under various bioreactor conditions. The systems analyses undertaken improve the understanding of driving forces of enzyme reactions, metabolite inhibitions, and adaptive cellular responses when yeast cells are under metabolic stresses. The new insights and principles are expected to facilitate the design and control of microbial pathways for effective bioproduction.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.