2D Enzyme Cascade Network with Efficient Substrate Channeling by Swinging Arms

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.

Document Details

Document Type
Pub Defense Publication
Publication Date
Jan 04, 2018
Source ID
10.1002/cbic.201700613

Entities

People

  • Hao Yan
  • Jinglin Fu
  • Minghui Liu
  • Shaun Wootten
  • Xiaodong Qi
  • Yan Liu
  • Yuhe Renee Yang

Organizations

  • Arizona State University
  • National Institutes of Health
  • Rutgers University
  • Rutgers University–Camden
  • United States Army Research Laboratory

Tags

Fields of Study

  • Biology

Readers

  • Electrical Engineering
  • Molecular and Cellular Biochemistry
  • Nanoscale Plasmonic Nanotechnology