RII-Track 4: Elucidating the Structure and Interactions of Complex Carbohydrates in Fungal and Plant Cell Walls via Dynamic Nuclear Polarization Solid-State NMR

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Non-Technical description The complex carbohydrate is a fundamental building unit of cell walls in a variety of organisms such as plants, fungi, bacteria and algae, which serve as a central source for bioenergy and biomaterials as well as a potential target of medical therapies. These polysaccharides are often polymorphic and disordered in their native state, however, due to technical limitations their functional structure remains under-investigated. This EPSCoR project will strengthen the collaborative efforts between Louisiana State University and National High Magnetic Field Laboratory (NHMFL), with the aim of developing a novel method to address several fundamental scientific questions that are related to polysaccharides. Specifically, the proposed research will advance understanding of the tendency and mechanism for carbohydrates to interact with other biomolecules as well as the building principles and conserved modules of cell walls. These molecular insights will guide the design and engineering of carbohydrate-rich composites to fulfill the diverse needs of the nation for better biofuel, material, and biomedicine. This collaborative project also allows scientists and students to gain frequent exposure to state-of-art magnet facilities, training at NHMFL and introduce a new technique that will build up the research capacity of Louisiana. Technical descriptionThis project aims to expand knowledge of carbohydrate research from soluble or crystalline molecules in purified components to the highly polymorphic, insoluble and disordered glycans in native cells. This goal will be achieved by integrating Dynamic Nuclear Polarization (DNP) technique and solid-state NMR methods to characterize the structure and the assembly of complex carbohydrates with minimal perturbation. By combing the sensitivity enhancement from DNP and the resolution improvement from spectral editing, the PI will be able to conduct site-specific investigations of polysaccharides in a complex whole-cell system. Structural characterizations will start from 13C,15N-labeled tissues and then progressively extend to unlabeled materials. This approach, once established, will alleviate the difficulty and expense that is typically associated with isotope-labeling, thus overcoming a long-standing barrier to large-scale structural elucidation. To enable rapid access to the results, a database will be established to document the NMR observables and structural restraints of polysaccharides as well as the diverse architecture of cell walls and other biomaterials. The proposed method and database has the potential to facilitate the modification of carbohydrate-rich composites to serve national demand for better bioenergy and materials. This collaborative project also provides a faculty and a postdoctoral fellow with the access to high-field NMR and DNP magnets at National High Magnetic Field Laboratory (NHMFL) and grants invaluable learning experiences through extensive interactions with well-established experts, thus stimulating the career development of next-generation scientists.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.