Genomics-Accelerated Discovery and Biosynthesis of Phosphonic Acid Natural Products - PROJECT SUMMARY The long-term goals of our research program are to discover new microbial natural products (NPs) of chemotherapeutic value, establish a mechanistic understanding of their underlying their biosynthesis and bioactivities, and to leverage these molecules, enzymes, and metabolic pathways for biomedical and biotechnological advancement. Evolved over millennia, NPs are chemically diverse small molecules produced from living organisms that exhibit with a wide range of useful inhibitory activities. NPs are a critical source of new drugs and often the initial chemical catalysts driving drug development. Phosphonic acids (Pn) are a privileged class of NPs with a prolific history of development and commercialization as antibiotics, antivirals, antimalarial, anticancer, and herbicidal agents. Universally defined by a carbon-phosphorous (C-P) bond, this moiety enables unique targeting of essential pathways in cellular metabolism thru chemical mimicry of enzyme substrates, products, and transition-state intermediates. These traits, combined with a near boundless collection of uncharacterized biosynthetic gene clusters for Pn encoded in microbial genomes and metagenomes, suggest that a significant reservoir of novel compounds remain for this important NP class. Genome mining is a proven method to accelerate discovery of NP leads through prediction of their pathways, molecules and activities from biosynthetic gene cluster sequence. However, fundamental gaps in our biosynthetic and chemical understanding of Pn NPs significantly limits the predictive power for this compound class. Thus, universal genetic, biochemical, and bioactivity principles of Pn NPs must be established to fully exploit their encoded genomic potential. In this proposal, we propose two research directions to address these goals. Direction 1 seeks to answer challenging, foundational questions central to Pn biosynthesis that will generate conceptual innovations necessary to improve their discovery. Using our expertise in bioinformatics, molecular microbiology, biochemistry, and chemical analysis, we will define essential characteristics of C-P bond enzymes, discover of new mechanisms C-P bond formation, and elucidate pathways for new Pn NPs. In Direction 2 our expertise in genome mining, NP isolation, and structure elucidation will be combined to isolate new inhibitory Pn NPs from diverse biosynthetic gene clusters of marine, soil dwelling, and plant associated microbes predicted to encode novel chemistries. Both directions afford natural integration throughout the project, as biosynthetic findings will support genome mining of new NPs, and newly uncovered molecules will drive studies that illuminate previously unknown paradigms in biosynthesis. Outcomes of this research program are expected to significantly advance Pn drug discovery thru refinements of their biosynthetic landscape and with the discovery of new NPs with potential therapeutic value.
