CAREER: Investigating Non-heme Iron Enzyme Catalyzed Nitrile Formation Mechanisms in Etoposide and Other Natural Product Biosynthetic Pathways

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Enzymes are biological catalysts that increase the speed of chemical reactions in the cell. Redirecting and repurposing enzymes to produce desirable novel chemical products is a challenge but could have significant industrial rewards. With this award, the Chemistry of Life Processes Program in the Chemistry Division is funding Dr. Wei-chen Chang from North Carolina State University to repurpose specific iron-containing enzymes to speed-up a variety of chemical reactions that these proteins do not typically perform. Recent computational and mechanistic advances point to opportunities to use these enzymes to "bio-synthesize" compounds of industrial interest, and to understand the fundamental principles of how to control such reactions. Dr. Chang's studies use a "substrate decoy", in which the standard starting compound is replaced by a new reactant to change the type of chemistry catalyzed by the enzyme. For this project, the iron-containing enzymes are reprogramed to alter their functions. Where they used to break down molecules, they now add chemical groups to create new compounds. The research plan is integrated with educational activities to inspire the next generation of students to pursue interdisciplinary research at the interface between chemistry and biology. The "science express" outreach activity uses a simple biochemical reaction that turns ordinary sugar into a blue dye (the "blue genes for blue jeans" hands-on experience) to show students how chemical biology impacts the world around them. These activities engage underrepresented K-12 students from diverse cultural, racial, and socioeconomic backgrounds in rural areas of North Carolina. This research project aims to develop a substrate decoy approach to redirect the reactions catalyzed by mononuclear non-heme iron and 2-oxoglutarate dependent (Fe/2OG) while maintaining their catalytic efficiencies. Dr. Chang's goal is to explore the potential of utilizing this approach to biosynthesize industrially important molecules. The research objective is to redirect the reaction outcome of these enzymes from hydroxylation to nitrile group installation. A complementary objective is to elucidate, through various chemical probing and kinetic approaches, the mechanism by which Fe/2OG hydroxylases catalyze this non-native reaction. Dr. Chang's educational objectives include developing an undergraduate course in enzyme kinetics that integrates chemical and biochemical concepts through active learning methods. In addition, his group has developed an easy-to-understand enzyme catalyzed reaction ("blue genes for blue jeans") as part of the science express outreach program. This program, established in collaboration with local teachers, introduces middle school students to exciting research in chemical biology.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.