CAREER: Bacterial Cytochrome P450 Enzymes and their Redox Partners

About This Grant

With the support of the Chemistry of Life Processes (CLP) program in the Division of Chemistry, Professor Jeffrey Rudolf of the University of Florida is studying a family of oxidative enzymes within the context of organic molecules made by bacteria. Natural products are small organic molecules that are made by organisms in nature. Natural products have profound impacts on society as commercially, agriculturally, and pharmaceutically relevant chemicals; they also serve as important probes for understanding fundamental processes in chemistry and biology. Cytochrome P450 enzymes (P450s), best known for their importance in human health, perform key enzymatic modifications of natural products in a variety of organisms including bacteria. The project aims to discover new natural products through the identification of unique P450s within the genomes of target bacteria and to understand the molecular interactions between P450s and their redox partners, proteins that are required for P450s functionality. This pursuit will allow graduate and undergraduate students to gain valuable training in bioinformatics, molecular biology, bacterial genetics, natural products chemistry, biochemistry, and analytical chemistry techniques. The project will also establish an Undergraduate Mentoring and Career Development Program (UMCDP) for select primarily undergraduate institutions (PUIs) and provide PUI students summer research experience opportunities in the Rudolf Lab. The goals of these programs are to increase awareness of career opportunities in science, provide a route to mentored research opportunities and graduate programs, and educate students in modern research topics and advancements. P450s are one of the most accomplished ‘chemists’ employed by nature and the study of microbial P450s reveals their diverse roles in nature, expands their catalytic repertoire, and exposes their potential for biotechnological applications. With the explosion of bacterial genomes now available, it is evident that our understanding and use of P450 catalysis is limited. This proposal seeks to achieve the following objectives: (i) discover novel natural products with unique P450 functionalities, (ii) build a foundation of sequence-structure-function knowledge through the characterization of P450s, and (iii) identify native redox partners of bacterial P450s. These goals will be accomplished through an interdisciplinary approach including genome mining, metabolomics and transcriptomics of native and genetically engineering bacteria, and functional characterization of P450s using genetic knockouts and a variety of biochemical and biophysical techniques for in vitro reconstitution. This proposal will make fundamental contributions to natural products biosynthesis and to understanding the general principles of P450 enzymology. In addition, synthetic and biosynthetic chemists who aim to functionalize unactivated C–H bonds or use P450s as biocatalysts will benefit from the results generated by this project. 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.