Date of Award
5-2025
Document Type
Thesis
Degree Name
Bachelor of Science
Department
Biochemistry & Molecular Biol.
First Advisor
Dr. Jonathan Dattelbaum
Second Advisor
Dr. Julie Pollock
Abstract
Members of the periplasmic binding protein superfamily are involved in the selective passage of ligands through bacterial cell membranes. The hyperthermophilic eubacterium Thermotoga maritima encodes a putative periplasmic branched chain amino acid binding protein (TM1135). The gene was cloned into pET21a(+), overexpressed in Escherichia coli, purified using Ni-NTA chromatography, and confirmed with SDS-PAGE and MALDI mass spectrometry. Biophysical characterization of the protein was performed with fluorescence spectroscopy, absorbance spectroscopy, and circular dichroism experiments to investigate structural stability and ligand binding ability. Using circular dichroism, we demonstrate that TM1135 binds to the branched chain amino acids isoleucine, leucine, and valine, and is stable to both temperature and denaturing agents. Similar to other periplasmic binding proteins, TM1135 may serve as a scaffold for future biosensor designs due to its thermodynamic and chemical stability.
Recommended Citation
Marsicano, Sky, "Biophysical Characterization of Thermophilic Branched-Chain Amino Acid Periplasmic Binding Protein from Thermotoga maritima" (2025). Honors Theses. 1840.
https://scholarship.richmond.edu/honors-theses/1840
