Implementing Multi-Enzyme Biocatalytic Systems Using Nanoparticle Scaffolds

Abstract

Interest in multi-enzyme synthesis outside of cells (in vitro) is becoming far more prevalent as the field of cell-free synthetic biology grows exponentially. Such synthesis would allow for complex chemical transformations based on the exquisite specificity of enzymes in a "greener" manner as compared to organic chemical transformations. Here, we describe how nanoparticles, and in this specific case-semiconductor quantum dots, can be used to both stabilize enzymes and further allow them to self-assemble into nanocomplexes that facilitate high-efficiency channeling phenomena. Pertinent protocol information is provided on enzyme expression, choice of nanoparticulate material, confirmation of enzyme attachment to nanoparticles, assay format and tracking, data analysis, and optimization of assay formats to draw the best analytical information from the underlying processes.

Keywords: Assay; Cascade; Catalysis; Enzyme; Kinetics; Michaelis–Menten; Multi-Enzyme; Nanoparticle; Quantum dot; Spectrophotometry; Synthetic Biology.