Abstract
Cellular functions rely on a series of organized and regulated multienzyme cascade reactions. The catalytic efficiencies of these cascades depend on the precise spatial organization of the constituent enzymes, which is optimized to facilitate substrate transport and regulate activities. Mimicry of this organization in a non-living, artificial system would be very useful in a broad range of applications—with impacts on both the scientific community and society at large. Self-assembled DNA nanostructures are promising applications to organize biomolecular components into prescribed, multidimensional patterns. In this review, we focus on recent progress in the field of DNA-scaffolded assembly and confinement of multienzyme reactions. DNA self-assembly is exploited to build spatially organized multienzyme cascades with control over their relative distance, substrate diffusion paths, compartmentalization and activity actuation. The combination of addressable DNA assembly and multienzyme cascades can deliver breakthroughs toward the engineering of novel synthetic and biomimetic reactors.
Highlights
Multistep enzyme pathways play critical roles in cellular metabolism that produces biomolecules and harvests energy for sustaining and propagating living systems
We summarize and discuss the recent progress in the field of DNA scaffold-directed assembly of multienzyme reactions, including proximity assembly, confinement, biomimetic substrate channeling and regulation circuits, as well as bioconjugation techniques of hybrid DNA–protein structures
The combination of DNA nanostructures with enzyme–DNA conjugation provides an efficient approach for engineering artificial biomolecule complexes with finely controlled spatial arrangement and confinement
Summary
CH Chlorohexane DSS Disuccinimidyl suberate DTPC DNA-templated protein conjugation DX Double-crossover DNA tile G6PDH Glucose-6-phosphate dehydrogenase GOx Glucose oxidase HRP Horseradish peroxidase IDE Inhibitor–DNA–enzyme LDH Lactate dehydrogenase MDH Malic dehydrogenase MTG Microbial transglutaminase NHS N -hydroxysuccinimide NiR Nitrite reductase ORBIT Origami-rotor-based imaging and tracking Paz Pseudoazurin Pg Plasminogen PLP Pyridoxal 5′-phosphate SK Streptokinase SMCC Succinimidyl-4-(N-maleimidomethyl) cyclohexane-1-carboxylate SPDP Succinimidyl 3-(2-pyridyldithio) propionate STVs Streptavidins TAL Transcription activator-like
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