Spatially Organized Enzymes Drive Cofactor-Coupled Cascade Reactions

Tien Ngo Anh, Eiji Nakata, Masayuki Saimura, Takashi Morii

Research output: Contribution to journalJournal articleResearchpeer-review

Abstract

We report the construction of an artificial enzyme cascade based on the xylose metabolic pathway. Two enzymes, xylose reductase and xylitol dehydrogenase, were assembled at specific locations on DNA origami by using DNA-binding protein adaptors with systematic variations in the interenzyme distances and defined numbers of enzyme molecules. The reaction system, which localized the two enzymes in close proximity to facilitate transport of reaction intermediates, resulted in significantly higher yields of the conversion of xylose into xylulose through the intermediate xylitol with recycling of the cofactor NADH. Analysis of the initial reaction rate, regenerated amount of NADH, and simulation of the intermediates' diffusion indicated that the intermediates diffused to the second enzyme by Brownian motion. The efficiency of the cascade reaction with the bimolecular transport of xylitol and NAD(+) likely depends more on the interenzyme distance than that of the cascade reaction with unimolecular transport between two enzymes.
Original languageEnglish
JournalJournal of the American Chemical Society
Volume138
Issue number9
Pages (from-to)3012-3021
Number of pages10
ISSN0002-7863
DOIs
Publication statusPublished - 2016
Externally publishedYes

Keywords

  • CHEMISTRY,
  • DNA-ORIGAMI STRUCTURES
  • MULTIENZYME SYSTEMS
  • METABOLIC ENZYMES
  • CRYSTAL-STRUCTURE
  • IN-VIVO
  • SCAFFOLDS
  • XYLOSE
  • NANOSTRUCTURES
  • COMPLEXES
  • DIFFUSION
  • Chemistry (all)
  • Catalysis
  • Biochemistry
  • Colloid and Surface Chemistry
  • Brownian movement
  • Enzymes
  • Reaction kinetics
  • Sugar substitutes
  • Artificial enzymes
  • Cascade reactions
  • DNA-binding protein
  • Enzyme molecules
  • Metabolic pathways
  • Specific location
  • Systematic variation
  • Xylose reductase
  • Reaction intermediates

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