Presentation Title

Probing Cytochrome P450 Biocatalysis via Anaerobic Redox Titrations

Faculty Mentor

Andrew K. Udit

Start Date

17-11-2018 8:30 AM

End Date

17-11-2018 10:30 AM

Location

CREVELING 57

Session

POSTER 1

Type of Presentation

Poster

Subject Area

biological_agricultural_sciences

Abstract

Cytochrome p450 is a versatile enzyme present in all kingdoms of life and can catalyze a variety of commercially relevant reactions to make new detergents, fuels, and drugs. A challenge to making the catalyst commercially available is that in native conditions cytochrome p450 requires NADP(H), which is hard to isolate and very expensive. The goal of this project is to supply the enzyme with the necessary electrons with common electricity through a surfactant-covered electrode and utilize the reaction-inducing ability of cytochrome p450 for industrial scale use. The p450 enzyme has the potential to be a significantly cheaper and greener catalyst replacement for current industrial scale reactions. However, we have found that the enzyme-electrode interaction alters the biophysical properties of p450, which renders it inactive. To understand the nature of the enzyme-electrode interaction and have cytochrome p450 perform with natural catalytic activity, we performed reduction-oxidation (redox) titrations to measure the redox potential and absorbance using UV-Vis spectroscopy. We were able to fit the electrochemical and spectroscopic data to the Nernst equation and obtain a relatively consistent Eo value. We will proceed to working with surfactants and modifying the ratio of surfactant to protein to reach catalysis rates comparable to in vivo while continuing to test the changes on the redox potential.

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Nov 17th, 8:30 AM Nov 17th, 10:30 AM

Probing Cytochrome P450 Biocatalysis via Anaerobic Redox Titrations

CREVELING 57

Cytochrome p450 is a versatile enzyme present in all kingdoms of life and can catalyze a variety of commercially relevant reactions to make new detergents, fuels, and drugs. A challenge to making the catalyst commercially available is that in native conditions cytochrome p450 requires NADP(H), which is hard to isolate and very expensive. The goal of this project is to supply the enzyme with the necessary electrons with common electricity through a surfactant-covered electrode and utilize the reaction-inducing ability of cytochrome p450 for industrial scale use. The p450 enzyme has the potential to be a significantly cheaper and greener catalyst replacement for current industrial scale reactions. However, we have found that the enzyme-electrode interaction alters the biophysical properties of p450, which renders it inactive. To understand the nature of the enzyme-electrode interaction and have cytochrome p450 perform with natural catalytic activity, we performed reduction-oxidation (redox) titrations to measure the redox potential and absorbance using UV-Vis spectroscopy. We were able to fit the electrochemical and spectroscopic data to the Nernst equation and obtain a relatively consistent Eo value. We will proceed to working with surfactants and modifying the ratio of surfactant to protein to reach catalysis rates comparable to in vivo while continuing to test the changes on the redox potential.