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.