Title

Improving the Synthesis of an Unnatural Fluorescent Amino Acid

Faculty Sponsor

Dr. Thomas Goyne

College

Arts and Sciences

Discipline(s)

Biochemistry

ORCID Identifier(s)

Stephanie Moore 0000-0003-4295-4884; Elliee Guido 0000-0002-7829-1771

Presentation Type

Poster Presentation

Symposium Date

Spring 4-28-2022

Abstract

The long-term goal of this project is to more efficiently synthesize an unnatural fluorescent amino acid, 3-[7-nitro-2,1,3-benzoxadiazol-4-yl]-L-alanine, diazole for short. This fluorescent amino acid can be incorporated into a transparent protein, thus creating a "glow-in-the-dark" protein. A biocatalysis pathway is being explored to synthesize this diazole amino acid. Specifically, we used the enzyme glutathione S-transferase from the cyanobacterium T. elongatus to catalyze the key nucleophilic aromatic substitution reaction. The gene for a His-tagged variant of this enzyme was inserted into a plasmid which was used to transform E. coli. Following protein expression, the cells were lysed and the enzyme was isolated using a nickel chelate column. Enzyme kinetics are monitored using LC-MS.

Biographical Information about Author(s)

Stephanie Moore is from Hobart, Indiana and will graduate from Valparaiso University in 2022. With a major in Biochemistry, Stephanie plans to enter into industry, and eventually obtain a PhD in Nutritional Biochemistry.

Elliee Guido is from Whiting, Indiana and will graduate from Valparaiso University in Spring of 2022 with a B.S in Chemistry. Elliee plans to go into the Chemistry work field, and then soon enter the medical field to become an anesthesiologist.

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