Integrated omics approach to identifying mechanisms of evolved tolerance to biobased chemical products

We propose to employ EMSL resources to identify the functional mechanisms for chemical tolerance in E. coli isolates that have been evolved to grow in high concentrations of chemicals that are of interest for production as bulk biochemicals or biofuels. The chemical-tolerant strains to be investigated were
obtained from a series of adaptive laboratory evolutions performed at the Center for Biosustainability (CFB). In FY2016, we specifically focused on the functional effects of cell wall and membrane-related mutations in a small set of strains. In FY2017, our aim is to characterize additional re-sequenced isolates, including those that do not have obvious cell wall or membrane-related mutations, through compositional profiling of membrane and cytoplasmic proteins, lipids, and intracellular metabolites using mass spectrometry. Additionally, transcriptional profiling of these strains is proposed through RNA sequencing. A further focus will be placed on isolates that exhibit increased endogenous production of the chemical. This information is expected to generate biological knowledge that will close the gap between genotype and phenotype.