(663b) Recombinant Production and Purification of Green Fluorescent Protein (GFP)-Fused Metal Binding Protein for Palladium Nanoparticle Synthesis

In lieu of chemical and physical methods, biologically guided synthesis is increasingly used as a cost-effective method for the fabrication of nanoparticles (NPs). Recently, a palladium metal binding sequence Pd4 (TSNAVHPTLRHL) has been demonstrated to be instrumental in controlling the production of palladium (Pd) nanoparticles. This work aims to provide an analysis of the catalytic turnover frequency of NPs derived from (Pd4)₃-GFPuv and correlate these values to the level of recombinant purity. An E. coli expression system was employed for expression of three copies of palladium binding sequence fused with green fluorescence protein (GFP). This protein was purified with chromatography from cell lysate. Transmission electron microscopy (TEM) was utilized to characterize the synthesized NPs from clarified lysate and pure protein sample, and the catalytic property of fabricated NPs from the purified protein was analyzed by a model Suzuki-Miyaura coupling reactions. It was observed that the purified metal binding protein had are three times higher turnover frequency compared to the crude lysate previously published.