Biochemical Analysis of Cytochromes P450 (CYPs) in the Betalain Biosynthesis Pathway of Beta vulgaris

Alex Bare

Abstract

Cytochromes P450 (CYPs) are a class of enzymes involved in metabolism, maintaining homeostasis, antioxidation, among other important biochemical functions. CYP76, specifically, plays a vital role in betalain pigment biosynthesis in beets. The three-dimensional (3D) structure of CYP76 can provide a molecular basis regarding its function and potential applications. Protein expression via molecular biology techniques such as plasmid transformation into competent Escherichia coli (E. coli) cells, inoculation, IPTG induction, and protein purification were used in this research with the intention of crystallizing the protein in an effort to study its 3D structure. Protein analysis techniques, specifically SDS-PAGE, are used to analyze the purity of the purified protein. This research is still in its early stages, but recent SDS-PAGE results suggest that there may be solubility problems, impurities, or low protein content.

Faculty Advisor: Soon Goo Lee

Acknowledgment: Shanyyce Bailey, Jake Barras, Kristy Witte, Kerstin Schmiederer

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Alex Bare

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Email: ajb2268@uncw.edu