Date of Graduation
5-2023
Document Type
Thesis
Degree Name
Bachelor of Science in Chemistry
Degree Level
Undergraduate
Department
Chemistry & Biochemistry
Advisor/Mentor
Thallapuranam, Suresh
Committee Member/Reader
Adams, Paul
Committee Member/Second Reader
Ceballos, Ruben
Abstract
Fibroblast growth factors (FGFs) are a family of signaling proteins with diverse biological functions. Fibroblast growth factor 19 (FGF19) regulates several vital physiological processes, including bile acid, glucose, and lipid homeostasis. However, its therapeutic potential is limited by its structural instability. The purpose of this study is to express, purify, and characterize recombinant FGF19 to build the foundation for the creation of stable mutants, which can be used to treat a variety of common diseases including type 2 diabetes, atherosclerosis, and nonalcoholic fatty liver disease. The parameters for overexpression in Escherichia coli were optimized according to optical density, inducer concentration, and post-induction incubation period. The protein was purified using immobilized metal affinity chromatography, and its secondary and tertiary structures were studied using fluorescence spectroscopy and far-ultraviolet circular dichroism spectroscopy, respectively. The protein was found to have a melting temperature of 47.72 °C and to denature at a urea concentration of 1 M. This data will offer a standard against which to compare future variants designed to increase the protein’s stability and enhance its bioactivity.
Keywords
Fibroblast growth factors, recombinant protein characterization, drug development
Citation
Ford, E. (2023). Expression, Purification, and Characterization of Fibroblast Growth Factor 19. Chemistry & Biochemistry Undergraduate Honors Theses Retrieved from https://scholarworks.uark.edu/chbcuht/39