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

Washington University, Saint Louis MO

Investigators

Abstract

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.