RUI: Exploiting biophysical techniques to understand and manipulate the mechanism and activity of electrode-bound cytochrome P450

Occidental College, Los Angeles CA

Investigators

Abstract

The Chemistry of Life Processes Program in the Chemistry Division of the National Science Foundation is awarding Dr. Andrew K. Udit of Occidental College funds to study the mechanism and activity of electrode-bound cytochrome P450. The cytochromes P450 are ubiquitous enzymes that can catalyze diverse chemical reactions. Reproducing the P450 activity in vitro depends on having an efficient way to deliver electrons to the enzyme. The proposed experiments will use biophysical techniques to study electron transfer and the reactivity of P450 immobilized on a variety of electrode surfaces. The results of this research will be useful to develop a fully competent, catalytic P450 electrode, which in turn has potential industrial applications. The research will be conducted by undergraduate and high school students and the results of the research will be used to generate course materials for undergraduate laboratories. While P450-electrode electronic coupling can be achieved often with rapid electron transfer, catalysis akin to the native system is not easily realized; this is especially true for bacterial variants, whose stability, substrate plasticity, and rapid rates make them ideal candidates for in vitro applications. Dr. Udit will use biophysical methods including isothermal calorimetry, bio-layer interferometry, and electrochemistry to study the mechanism and activity of the P450 enzyme on a variety of electrode surfaces (e.g., trapped in films on- or covalently-attached to- the electrode surface) and solvent environments. The long term aim of the research is to improve the activity of electrode-coupled P450, which is of considerable practical interest for applications in areas such as biosensing and biocatalysis.