Nanoparticles have emerged as important tools in biomedicine, with numerous clinical applications including contrast agents in bioimaging and drug and gene delivery carriers. However, thorough investigation of their implications on biomolecular systems remain underexplored. Developing a functional knowledge of these interactions can be used to generate nanoparticles compatible with biological environments. The primary interest of this project has been to synthesize nanoparticles and study their impacts on the structure and function of an important family of enzymes known as aminoacyl-tRNA synthetases, which are responsible for attaching amino acids to a cognate tRNA molecule. Gold and silver nanoparticles, with ranging differences in core size and functionalization groups, were synthesized and purified using a variety of methods. Two unique biocompatible ligands were utilized, including citrate and 2-[2-(2-Mercaptoethoxy)ethoxy]ethanol. These nanoparticles were used to examine their impact on the conformation and function of E. coli prolyl-tRNA synthetase using intrinsic fluorescence spectroscopy and enzyme kinetics. The present study demonstrated that gold nanoparticles, regardless of diameter or functionalization, do not influence the conformation and function of E. coli prolyl-tRNA synthetase. On the other hand, silver nanoparticles have impact on the protein conformation but not the enzymatic function.