The Synthesis of an Unnatural Fluorescent Amino Acid
Dr. Thomas Goyne
Arts and Sciences
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.
Niemeyer, Cassandra; Kurth, Esteban; and Gaskill, Taylor, "The Synthesis of an Unnatural Fluorescent Amino Acid" (2021). Symposium on Undergraduate Research and Creative Expression (SOURCE). 953.