Enzymatically Active DNA-Protein Nanogels with Tunable Cross-Linking Density

Hybrid DNA-protein nanogels represent potential protein vectors and enzymatic nanoreactors for modern biotechnology. Here, we describe a new, easy, robust method preparation of tunable with controllable size density. For this purpose, polymerase chain reaction is used to prepare highly biotinylated DNA as soft biopolymeric backbone, which can be efficiently cross-linked via streptavidin-biotin binding. This approach enables us control both the density resulting not only by adjusting amount cross-linking streptavidin but also using different rates biotinylation. gives DNA-streptavidin ranging from 80 nm, most compact state, up 200 nm. Furthermore, streptavidin-enzyme conjugates allows straightforward one-pot incorporation enzymes during nanogels. Monoenzymatic multienzymatic have been obtained in manner, their catalytic activities characterized. All tested (alkaline phosphatase (AP), horseradish peroxidase (HRP), ?-galactosidase (?Gal)), incorporated individually or coupled manner (glucose oxidase (GOx)-HRP cascade), were shown remain functional. The AP ?Gal unchanged while that HRP was slightly improved inside We demonstrate that, HRP, it DNA-to-enzyme ratio physical functionalized responsible improvement its activity.