1.Metabolomics analysis of B. subtilis strain 168 in the presence and absence of n-butanol. This aim is to use untargeted metabolomics strategies to measure lipophilic metabolite levels in B. subtilis incubated with and without n-butanol to enable the discovery of lipophilic metabolites potentially responsible for improving tolerance of B. subtilis in n-butanol.
2. Characterization of metabolites whose levels are altered in n-butanol-treated samples. In this aim, we propose to use various bioinformatics, chemical, analytical, and chromatographic techniques to characterize metabolite changes in aim 1, especially those elevated in n-butanol-treated samples.
3. The correlation between elevating metabolites and n-butanol tolerance of B. subtilis. The tolerance of B. subtilis toward n-butanol exposure might correlate to the presence of lipophilic metabolites elevated in response to n-butanol. This aim is designed to test this hypothesis by investigating the growth of B. subtilis in the presence of n-butanol and the elevating molecules characterized in aim 2.
