Date(s) - 11/21/2023
9:00 am - 10:00 am
Carl Denard, Ph.D.
Department of Chemical Engineering
University of Florida
Title: Leveraging yeast ER sequestraon to study and engineer post-translaonal modificaon enzymes.
Enzymes that catalyze post-translaonal modificaons of pepdes and proteins (PTMenzymes) – proteases, protein ligases, oxidoreductases, kinases, and other transferases – are foundaonal to our understanding of health and disease and empower applicaons in chemical biology, synthec biology, and biomedicine. To fully harness their potenal, there is a crical need to decipher their enzymac and biological mechanisms, develop molecules that can probe and reprogram them, and endow them with improved and novel funcons. These objecves are conngent upon implementaon of high-throughput funconal screens and selecons that interrogate large sequence libraries to isolate desired PTM-enzyme properes.
In this seminar, I will talk about the development and applicaon of yeast ER sequestraon 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 evoluon to modify biochemical properes, by funconal interacons with a selecve ligand, and through substrate engineering and profiling. Specific examples will be described, including 1) engineering the substrate specificity of a human protease therapeuc, a so-called disaggregase, 2) developing a plaorm 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 mutaonal landscape of the ER targeng protein SRP54. Lastly, I will emphasize how we use principles of chemical engineering to improve yeast ER cell factories.
Dr. Denard is currently an Assistant Professor in the Chemical Engineering Department at the University of Florida. Originally from Hai, 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 cooperave one-pot chemoenzymac reacons. Dr. Denard later completed a postdoctoral fellowship in the lab of Prof. Brent Iverson at the University of Texas at Ausn establishing high-throughput plaorms for engineering the substrate specificity of proteases to enable their use as protein therapeucs. The Denard lab at UF focuses on protein and biomolecular engineering. The primary research focus is to develop tools to reprogram post-translaonal modificaon enzymes for biomedical research, biotechnology, chemical and synthec biology. For his research efforts, Dr. Denard recently received the NIH MIRA award (2022) and the NSF CAREER award (2023).