一种将脲酶位点特异性锚定到DNA折纸结构并保持活性的方法
A method for site-specifically tethering the enzyme urease to DNA origami with sustained activity
AI总结：
本文提出一种将单链DNA偶联至脲酶并保持其活性的方法，实现脲酶以高位点特异性锚定到DNA折纸纳米结构上，为研究酶功能及生物技术应用提供新工具。
摘要：
将酶附着到纳米结构上已被证明有助于在受控条件下研究酶的功能，并催生了新技术。通常，酶-纳米结构复合物的效用和兴趣在于酶活性如何受到酶在空间中排列方式的影响。因此，在保持酶活性的同时将酶偶联到纳米结构上至关重要。本文提出了一种将单链DNA偶联到脲酶上同时保持酶活性的方法。我们展示了成功偶联的证据，并量化了影响偶联产率的变量。我们还表明，偶联后酶的活性与天然状态相比未发生变化。最后，我们展示了将脲酶以高位点特异性锚定到DNA折纸纳米结构上的方法。用具有酶活性的脲酶装饰纳米结构可能有助于研究甚至利用脲酶在从生物技术到软物质物理等学科中的功能。本文提出的技术将使这些领域的研究人员能够在不破坏酶功能的情况下修饰酶，从而对其行为和效用进行更深入的研究。
英文摘要：
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.
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