Functional Annotation of Natural Products: Methods and Applications to Marine Microbiota
Natural products have made dramatic contributions to a significant portion of FDA approved therapeutics. While many of these secondary metabolites have been exhaustively studied, the true purpose of these molecules is often left unresolved. Mechanism of action (MOA) annotation remains one of the largest bottlenecks in drug discovery. In addition, the source of natural products affects chemical diversity and metabolites isolated from microbiota are an emerging source of antimicrobials. They are believed by many to modulate host health, affecting predisposition to disease and pathogen invasion. Through the application of unbiased screening methods to novel sources, a more comprehensive understanding of the global effects of natural products can be achieved. This dissertation will present the implementation of an image based cytological profiling platform for the isolation of compounds with a specific, predefined mode of action, while simultaneously characterizing all bioactive constituents isolated from the crude extract. The development of a training set independent genetic assay in Saccharomyces cerevisiae will be discussed. In addition, the application of image based and genetic techniques to a new compound, phocoenamicin, isolated from marine mammal microbiota demonstrates the importance of identifying both new sources of natural products while simultaneously developing new tools to characterize these compounds, generating a more comprehensive image of what natural products are and what they are capable of.