Grant: Research Tools and Instruments - Category 1 (<$150,000)

This application is for a high performance liquid chromatography (HPLC) instrument to be used in support of an NSERC Discovery Grant program. The research program involves identifying biologically active small molecule natural products of microbial origin. The HPLC will be used to identify and purify small molecule metabolites from cultures of microorganisms that, for example, solubilize inorganic phosphate or produce biologically active phosphonates. The ability of microorganisms to solubilize inorganic phosphate complexes in soil via production of organic acids has been known for some time, but the biochemistry of this process is not well understood and other mechanisms are thought to exist. In addition, phosphonate small molecules possessing biological activity have found use as herbicides and antibiotics, yet relatively few natural phosphonates have been discovered and little is understood about their microbial biosynthesis. In order to learn more about these processes and identify novel molecules, massive collections of DNA from different environments will be expressed in bacteria and screened for functions such as phosphate solubilization and phosphonate production. The bacteria possessing the environmental DNA (eDNA) capable of the desired function will be cultured on a larger scale and characterized by HPLC. This is a crucial instrument for identifying small molecules from microbial cultures and for elucidating their biosynthetic pathways. Identification of novel mechanisms of phosphate solubilization will contribute to more efficient use of non-renewable phosphate fertilizer, and new phosphonate compounds will have potential impact in agriculture and medicine. The instrument will be an essential tool for the research program and will contribute significantly to student training.