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使用DNA框架纳米导体构建双酶复合体,以提高酶级联催化效率

Nan Cao^1,2, Shaopeng Wang¹, Fan Li¹

¹School of Chemistry and Chemical Engineering, Institute of Molecular Medicine Shanghai Key Laboratory for Nucleic Acid Chemistry and Nanomedicine, Renji Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai, 200127, P.R., China.

|February 14, 2024

概括

四面体DNA框架 (TDFs) 精确地定位酶,提高生物催化级联效率. 这项研究表明,TDFs通过控制酶的近距离,显著提高了氨酸氧化酶/马过氧化酶活性.

科学领域:

生物化学生物化学
纳米技术纳米技术
分子生物学分子生物学

背景情况:

酶级联反应对于细胞内转化至关重要,但由于溶液中的空间布局而受到限制.
控制酶的接近和形态是提高催化效率的关键.

研究的目的:

利用框架核酸 (FNA),特别是四面体DNA框架 (TDF),作为支架来精确控制酶的空间布局.
通过TDF介导的组装来增强萨尔科辛氧化酶 (SOx) /胡卜过氧化酶 (HRP) 酶级联系统的催化效率.

主要方法:

合成了四种类型的TDF纳米导体,用于编程从5.67nm到12.33nm的酶间距离.
化学修饰的SOx和HRP酶与单链DNA (ssDNA) 进行固定.
使用聚烯胺凝电泳 (PAGE),传输电子显微镜 (TEM) 和原子力显微镜 (AFM) 验证了双酶复合体的形成.

主要成果:

使用TDFs构建了四个具有不同酶距离的双酶复合体.
观察到与自由酶相比,所有TDF组装的酶复合体的催化活性显著增强.
鉴定TDF₂₆(SOx+HRP) 具有最高的相对酶级联催化活性,大约是自由状态酶的3.11倍.

结论:

TDF 作为有效的纳米导体,用于组装酶级联系统,优化空间组织.
使用TDFs精确控制酶间距离,大大提高了生物催化效率.
这种方法为设计各种应用的高效酶级联反应提供了一个有希望的策略.

关键词:

在DNA纳米制造器中.双酶复合体是一个双酶复合体.酶级联反应是一种酶级联反应.框架核酸核酸的框架间距控制间距控制间距控制

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