This proposal was co-funded by the Systems and Synthetic Biology program, Division of Molecular and Cellular Biosciences; the Chemistry of Life Processes Program, Division of Chemistry, and the Biocatalysis program of the Chemical, Bioengineering, Environmental, and Transport Systems Division.
Biocatalysis has emerged as an important environmentally friendly route for the production of green and sustainable chemicals. Among candidate biocatalysts, cytochrome P450s are of great interest due to their synthetic potential in the selective oxidation of unactivated C-H bonds, using molecular dioxygen and two reducing equivalents. The proposed work will expand the capability of this class of enzymes for the production of specific chemicals in a sustainable way and with selectivity not readily obtainable with conventional chemistry. The work will be done by undergraduate students at San Jose State University (SJSU), a predominantly undergraduate minority-serving institution located in the heart of the Silicon Valley.
Specifically, this RUI award supports the optimization of efficient light-driven hybrid P450 biocatalysts. This class of enzyme is, however, still underused in biotechnological applications due to the requirement for an electron providing reductase and an expensive NAD(P)H cofactor. Undergraduate students in the PI?s laboratory have engineered hybrid P450 BM3 biocatalysts, which contain strategically positioned Ru(II)-diimine photosensitizers capable of delivering the necessary electrons to their active sites and sustaining photocatalytic activity. This approach circumvents the need for the redox partner enzymes and the NAD(P)H cofactor. The proposed work will utilize a three-pronged approach to improve the performance of the light-driven biocatalysts by 1) expanding the scope of their photocatalytic activity towards non-natural substrates, 2) enhancing their reusability, stability, and activity through immobilization, and 3) combining the approach with photoredox catalysis to expand C-H bond functionalization using visible light excitation.
The proposed research and educational objectives will keep engaging a diverse group of undergraduate students and teachers while enhancing their educational experiences at SJSU and beyond. Authentic research experiments will be integrated into a capstone biochemistry lab course to enrich student learning and train them for successful future careers in STEM fields. In addition, a summer workshop will be held for high school teachers to provide hands-on training and enable the implementation of light-driven experiments in their high-school classrooms.
