Engineering 3-Dimensional Protein Scaffolds for Biocatalysts Assembly

In nature, enzymes are often assembled into complex structures that are more effective than individual enzymes in converting raw materials into useful products. Many biologically-derived scaffolds have been used for single and multi-enzyme assembly and have shown increased overall reaction rates in one- or two-dimensional configurations. Analogously, we hypothesize that the use of three-dimensional scaffolds could further enhance the performance of biocatalysts. The E2 core of the pyruvate dehydrogenase complex from Bacillus stearothermophilus is a genetically-modifiable 60-mer protein nanocage that is thermally stable at temperatures as high as 70°C. In this talk, we will discuss different approaches in pairing the E2 nanocage with a thermophilic endoglucanase, CelA from Clostridium thermocellum, resulting an enzyme assembly that benefit from both the synergy gained from clustering and stability at more enzymatically favorable high reaction temperatures. This E2-based approach offers a flexible platform upon which multi-enzyme cascade reactions can be assembled and studied at the nanoscale.