Widely existing multi-enzyme complexes (MECs) in nature exhibit highly efficient catalytic mechanism in cascaded reactions, which has inspired researchers to develop novel strategies to mimic MEC structure and function in vitro. It has been found that the spatial arrangement and orientation of multiple enzymes in artificial MECs plays a significant role in facilitating cooperative enzymatic activity. DNA hybridization provides unique effective control of the relative positions of different enzymes in multi-enzyme co-localization. It is also known that, compared to planar surfaces, a spherical geometry is more suitable for co-localization of multiple layers of enzymes due to lesser steric hindrance. To demonstrate this concept, glucose oxidase (GOX) and horseradish peroxidase (HRP) were co-localized onto polystyrene nanoparticles via specific DNA hybridization. In comparison with planar polystyrene films, it was shown that the nanoparticles provided higher hybridization efficiency. This behavior was also shown to be a function of particle size. Various capture DNA concentrations were compared and optimized. The co-localization of GOX and HRP were evidenced by FRET studies of the dyes labeling the two enzymes. In addition, different molar ratios of multiple enzymes in DNA directed co-localization were investigated. In summary, DNA directed co-localization of enzymes provides precise control over the relative positions of the enzymes to mimic MECs.

DNA directed multi-enzyme co-localization on nanoparticles

245th National Meeting (2013)

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