NSF Award
2300890
More bacteria have become resistant to current antibiotics. Natural compounds with antibiotic properties are assembled in a modular fashion inside cells. The modular nature allows for the design of hybrid pathways to produce non-natural molecules. The mechanisms of individual bond-forming reactions in an antibiotic molecule will be investigated. The intention is to develop a degree of insight that would enable the shuffling of building blocks to produce novel antibiotics. The concept is akin to molecular building blocks. In addition, this work will provide a foundation for training undergraduate students in conducting biotechnological research and communicating science to their community. <br/><br/>The project will attempt to uncover novel enzymatic mechanisms that facilitate production of unique natural products. Specifically, this work will elucidate mechanistic underpinnings of amino acid incorporation on polyketide scaffolds. The project focuses on the model molecule TLN-05220, which is an Echinosporamicin-type antibiotic with antibacterial activity against several strains of methicillin-resistant Staphylococcus aureus and vancomycin-resistant Enterococci (VRE), and cytotoxic activities against several human tumor cell lines. Individual gene functions in TLN-05220 biosynthesis will be identified using gene knockouts in the wild-type producing organism. Potential substrate binding sites and catalytic residues of amino acid-incorporating enzymes will be identified using homology modeling. Expression and characterization of individual enzymes will elucidate mechanism(s) of amino acid incorporation into natural product structures. The minimally necessary genes to produce TLN-05220 will be cloned and expressed in a heterologous host.<br/><br/>This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.
