A method for site-specifically tethering the enzyme urease to DNA origami with sustained activity

TL;DR

This paper introduces a method to conjugate the enzyme urease to DNA origami nanostructures while preserving enzymatic activity, enabling precise spatial arrangement for advanced biotechnological applications.

Contribution

The authors developed a novel technique for site-specific attachment of urease to DNA origami, maintaining enzyme activity and quantifying conjugation efficiency.

Findings

Successful conjugation of urease to DNA with high yield

Enzymatic activity remains unchanged after conjugation

High site-specific tethering of urease to DNA origami structures

Abstract

Attaching enzymes to nanostructures has proven useful to the study of enzyme functionality under controlled conditions and has led to new technologies. Often, the utility and interest of enzyme-tethered nanostructures lie in how the enzymatic activity is affected by how the enzymes are arranged in space. Therefore, being able to conjugate enzymes to nanostructures while preserving the enzymatic activity is essential. In this paper, we present a method to conjugate single-stranded DNA to the enzyme urease while maintaining enzymatic activity. We show evidence of successful conjugation and quantify the variables that affect the conjugation yield. We also show that the enzymatic activity is unchanged after conjugation compared to the enzyme in its native state. Finally, we demonstrate the tethering of urease to nanostructures made using DNA origami with high site-specificity. Decorating nanostructures with enzymatically-active urease may prove to be useful in studying, or even utilizing, the functionality of urease in disciplines ranging from biotechnology to soft-matter physics. The techniques we present in this paper will enable researchers across these fields to modify enzymes without disrupting their functionality, thus allowing for more insightful studies into their behavior and utility.