Development of Metabolite Profiling and Metabolomics Tools for Pseudomonas Taiwanensis

Strain development for the production of fuels and chemicals has traditionally relied on measurements of metabolite products titers to assess the performance of engineered strains. Today, improved metabolomics and targeted metabolite profiling methods make it possible to collect a broader range of measurements of the metabolic state of an organism. Despite of the challenges applying global metabolomics and targeted metabolite profiling methods, it is clear that their application provides significant benefits in both pathway discovery and synthetic biology. However, global metabolomics and metabolite profiling of strains is a challenging task due to the chemical and structural diversity of metabolites from polar to non-polar molecules. As a result of this diversity, methods used for quenching of metabolism and metabolite extraction significantly influence the number and intensity of metabolites detected by universal detection technologies.

In recent years, different sophisticated protocols including quenching with cold solvent mixtures and fast filtration have been developed for the quantification of intracellular metabolites of different strains especially yeast and bacteria. The direct application of these methods and other techniques to P. taiwanensis VLB120 strain, which is the only known solvent-tolerant Pseudomonas strain, is not well studied. Therefore, I am conducting a comprehensive examination of the metabolite profile of Pseudomonas VLB120 strains using different analytical techniques including gas and liquid chromatogram – mass spectrometry.