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

Area of Science:

• Biotechnology
• Nanotechnology
• Bioconjugation

Background:

• Hybrid DNA-protein nanogels are promising for applications as protein carriers and enzymatic nanoreactors.
• Developing methods for controlled synthesis of these nanogels is crucial for advancing biotechnology.

Purpose of the Study:

• To establish a simple, robust, and tunable method for preparing DNA-protein nanogels.
• To control nanogel size and density and to incorporate functional enzymes.

Main Methods:

• Utilized polymerase chain reaction (PCR) to create highly biotinylated DNA backbones.
• Employed streptavidin-biotin interactions for cross-linking DNA, controlling nanogel size (80-200 nm) and density.
• Incorporated enzymes (alkaline phosphatase, horseradish peroxidase, β-galactosidase) via streptavidin-enzyme conjugates in a one-pot procedure.

Main Results:

• Achieved tunable DNA-streptavidin nanogels with controllable size and density.
• Successfully incorporated and maintained the functionality of various enzymes, including monoenzymatic and multienzymatic systems.
• Observed unchanged activity for alkaline phosphatase and β-galactosidase, and a slight improvement for horseradish peroxidase, linked to nanogel density.

Conclusions:

• The developed method offers a versatile platform for creating functional DNA-protein nanogels.
• Enzyme activity within nanogels is influenced by physical density, not solely the DNA-to-enzyme ratio.
• These nanogels show significant potential as enzymatic nanoreactors in biotechnological applications.