2D Enzyme Cascade Network with Efficient Substrate Channeling by Swinging Arms. (4th January 2018)

• Record Type: Journal Article
• Authors: Yang, Yuhe R.; Fu, Jinglin; Wootten, Shaun; Qi, Xiaodong; Liu, Minghui; Yan, Hao; Liu, Yan
• Is Part Of: Chembiochem. Volume 19:Number 3(2018)
• Page Start: 212
• Page End: 216
• Publication Date: 2018-01-04
• Subjects: DNA nanotechnology -- enzyme cascade -- scaffold -- redox intermediate; Biochemistry -- Periodicals; Molecular biology -- Periodicals; Pharmaceutical chemistry -- Periodicals; 572
• DOI: 10.1002/cbic.201700613
• Languages: English
• ISSNs: 1439-4227

Abstract

Abstract: In living cells, compartmentalized or membrane‐associated enzymes are often assembled into large networks to cooperatively catalyze cascade reaction pathways essential for cellular metabolism. Here, we report the assembly of an artificial 2D enzyme network of two cascade enzymes—glucose‐6‐phosphate dehydrogenase (G6PDH) and lactate dehydrogenase (LDH)—on a wireframe DNA origami template. Swinging arms were used to facilitate the transport of the redox intermediate of NAD + /NADH between enzyme pairs on the array. The assemblies of 2D enzyme networks were characterized by gel electrophoresis and visualized by atomic force microscopy (AFM). The spatial arrangements of multiple enzyme pairs were optimized to facilitate efficient substrate channeling by exploiting the programmability of DNA origami to manipulate the key parameters of swinging arm length and stoichiometry. Compared with a single enzyme pair, the 2D organized enzyme systems exhibited higher reaction efficiency due to the promoted transfer of intermediates within the network. Abstract : Gaining momentum by swinging arms : We assembled an artificial 2D enzyme network of two related dehydrogenase enzymes on a wireframe DNA origami template to facilitate swinging of the redox intermediate substrate to enhance enzyme cascade activity. The 2D organized enzyme system exhibited higher reaction efficiency than single enzyme pairs due to promoted transfer of intermediates within the network.