The Synthesis of an Unnatural Fluorescent Amino Acid

Faculty Sponsor

Dr. Thomas Goyne


Arts and Sciences



Presentation Type

Poster Presentation

Symposium Date

Spring 4-29-2021


The long-term goal of this project is to develop a more cost-effective chemical synthesis of an unnatural fluorescent amino acid, 3-[7-nitro-2,1,3-benzoxadiazol-4-yl]-L-alanine. This molecule can be used to visualize a single “glow-in-the-dark” protein in an otherwise transparent living cell. We are investigating two synthetic routes: traditional organic chemistry and biocatalysis. The biocatalysis route utilizes the enzyme glutathione S-transferase from the thermophilic cyanobacterium, T. elongatus. For the organic reaction, nuclear magnetic resonance (1H NMR) spectroscopy is being used to monitor reaction kinetics with the goal of identifying the optimal solvent and reaction conditions. For the biocatalyzed reaction, a commercial vendor has synthesized the DNA that codes for the T. elongatus glutathione S-transferase gene and incorporated that gene into a suitable plasmid. We have used this plasmid to transform E. coli and have demonstrated expression of T. elongatus glutathione S-transferase. A plate reader has been used to monitor enzyme kinetics. We hope to develop a liquid chromatography-mass spectroscopy method that will allow us to monitor reaction kinetics for both the organic and biocatalysis routes.

Biographical Information about Author(s)

Cassandra is a senior Biochemistry major at Valparaiso University and will be entering medical research after graduation this spring.

Esteban is a senior Biochemistry major at Valparaiso University and will be hopefully entering the industry after graduating in the fall.

Taylor is a sophomore Chemistry major at Valparaiso University and is currently pursuing pre-med.

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