(512b) Multi-Enzyme Co-Localization with Nanoscale Pluronic–QD Micelles

To mimic the structure and functionality of multi-enzyme complexes, which are widely present in Nature, Pluronic^®-based nanomicelles were designed to co-localize multiple enzymes. To stabilize the micelles as well as to enable characterization of single enzyme immobilization and multi-enzyme co-localization using Förster Resonance Energy Transfer (FRET), quantum dots (QDs) were encapsulated into the polymeric micelles to form Pluronic^®-QD nanomicelles using a novel micro-reactor. Model enzymes, glucose oxidase (GOX) and horseradish peroxidase (HRP), were respectively labeled with fluorescent dyes. The results showed that FRET occurred between the QDs and dyes that labeled each type of enzyme in single enzyme immobilization as well as between the different dyes that served to label the enzymes. These observations were consistent with dynamic light scattering data. The enzymatic activity of single GOX or HRP was retained after immobilization. The optimized sequential co-localization of GOX and HRP was found to improve the overall conversion rate by approximately 100% compared to the equivalent amount of free enzymes in solution or a physical mixture of individual immobilized enzymes on nanoparticles. This study demonstrates a simple platform for multi-enzyme co-localization and a facile and effective strategy to characterize multi-enzyme immobilization and co-localization.