ChE: Leveraging yeast ER sequestration to study and engineer post-translational modification enzymes.


9:00 am-10:00 am
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HPNP 1404
1225 Center Drive
Gainesville, Florida 32611


Abstract: Enzymes that catalyze post-translational modifications of peptides and proteins (PTM-enzymes) – proteases, protein ligases, oxidoreductases, kinases, and other transferases – are foundational to our understanding of health and disease and empower applications in chemical biology, synthetic biology, and biomedicine. To fully harness their potential, there is a critical need to decipher their enzymatic and biological mechanisms, develop molecules that can probe and reprogram them, and endow them with improved and novel functions. These objectives are contingent upon implementation of high-throughput functional screens and selections that interrogate large sequence libraries to isolate desired PTM-enzyme properties.

In this seminar, I will talk about the development and application of yeast ER sequestration and screening to effectuate various high-throughput enzyme biochemical studies. We are applying this approach to reprogram PTM-enzymes in three ways: through protein engineering and directed evolution to modify biochemical properties, by functional interactions with a selective ligand, and through substrate engineering and profiling. Specific examples will be described, including 1) engineering the substrate specificity of a human protease therapeutic, a so-called disaggregase, 2) developing a platform to isolate protein-based modulators and screen small molecule inhibitors that can reprogram proteases, and 3) comprehensively profiling the substrate specificity of protein ligases, 4) and deciphering the mutational landscape of the ER targeting protein SRP54. Lastly, I will emphasize how we use principles of chemical engineering to improve yeast ER cell factories.

Bio: Dr. Denard is currently an Assistant Professor in the Chemical Engineering Department at the University of Florida. Originally from Haiti, Dr. Denard emigrated to the U.S. where he received his B.S. degree in Chemical Engineering from North Carolina State University. He then moved to the Chemical and Biomolecular Engineering department at the University of Illinois at Urbana-Champaign, where he obtained his PhD under the tutelage of Prof. Huimin Zhao. As a Dow Chemical fellow, his PhD thesis centered on developing cooperative one-pot chemoenzymatic reactions. Dr. Denard later completed a postdoctoral fellowship in the lab of Prof. Brent Iverson at the University of Texas at Austin establishing high-throughput platforms for engineering the substrate specificity of proteases to enable their use as protein therapeutics. The Denard lab at UF focuses on protein and biomolecular engineering. The primary research focus is to develop tools to reprogram post-translational modification enzymes for biomedical research, biotechnology, chemical and synthetic biology. For his research efforts, Dr. Denard recently received the NIH MIRA award (2022) and the NSF CAREER award (2023).


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Department of Chemical Engineering