Toward Rational Design of High-efficiency Enzyme Cascades

Zhang, Yifei; Hess, Henry

doi:10.1021/acscatal.7b01766

Cited by 183 publications

(211 citation statements)

References 117 publications

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“…Zhang and Hess have recently emphasized that the rational design of high-efficiency enzyme cascades will benefit not only from spatial proximity of cooperating enzymes but also from balanced stoichiometry 42 and Castelana et al 43 have already demonstrated that enzyme clustering accelerates the processing of intermediates through metabolic channeling. This type of compartmentalisation is manifested in the sequestration of the NADP(H) cofactor and hydroxyketone intermediates inside our all-enzyme hydrogels, which occurs even under continuous flow conditions.…”

Section: Discussionmentioning

confidence: 99%

Self-assembling all-enzyme hydrogels for biocatalytic flow processes

Peschke

Gallus

Bitterwolf

et al. 2017

Preprint

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______________________________________________________________We describe the construction of binary self-assembling all-enzyme hydrogels that are comprised entirely of two tetrameric globular enzymes, the stereoselective dehydrogenase LbADH and the cofactor-regenerating glucose 1-dehydrogenase GDH. The enzymes were genetically fused with a SpyTag or SpyCatcher domain, respectively, to generate two complementary homo-tetrameric building blocks that polymerise under physiological conditions into porous hydrogels. The biocatalytic gels were used for the highly stereoselective reduction of a prochiral diketone substrate where they showed the typical behaviour of the coupled kinetics of coenzyme regenerating reactions in the substrate channelling regime. They effectively sequestrate the NADPH cofactor even under continuous flow conditions. Owing to their sticky nature, the gels can be readily mounted in simple microfluidic reactors without the need for supportive membranes. The reactors revealed extraordinary high space-time yields with nearly quantitative conversion (>95%), excellent stereoselectivity (d.r. > 99:1), and total turnover numbers of the expensive cofactor NADP(H) that are amongst the highest values ever reported. ______________________________________________________________

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Section: Discussionmentioning

confidence: 99%

Self-assembling all-enzyme hydrogels for biocatalytic flow processes

Peschke

Gallus

Bitterwolf

et al. 2017

Preprint

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“…Reproduced from Grossi et al [114] under the terms and conditions of the Creative Commons Attribution 4.0 International License, copyright 2017, Springer Nature More interestingly, DNA nanoscaffolds have been found to affect the activities of the enzymes that were attached onto them. Figure 9 summarizes some of the DNA structures that have been reported to enhance enzyme activities [115], including a long dsDNA molecule (e.g. λDNA, ~ 1-2 fold improved activity) [116], a DNA structure binding to enzyme substrates (~ 1-2 fold) [117], a 2D rectangular DNA origami (~ 1-3 fold) [45], DNA nanocaged enzymes (~ 3-6 fold) [94,118] and DNAcrowded enzyme particles (~ 2-3 fold) [95].…”

Section: Enzyme Compartmentalization By Dna Nanocagesmentioning

confidence: 99%

DNA-Scaffolded Proximity Assembly and Confinement of Multienzyme Reactions

Wang

Liang

et al. 2020

Top Curr Chem (Z)

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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.

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Section: Dna Nanocage‐encapsulated Enzymesmentioning

confidence: 99%

“…Many recent studies have reported that enzyme activity was enhanced upon attachment to DNA nanostructures (Figure j), including a long double‐stranded DNA (dsDNA) molecule (e.g., λDNA, ≈1–2‐fold), a 2D rectangular DNA origami (≈1–3‐fold), a DNA scaffold binding to enzyme substrates (≈1–2‐fold), DNA nanocaged enzymes (≈3–6‐fold), and DNA‐crowded enzyme particles (≈2–3‐fold) . Several mechanisms have been proposed to interpret the enhanced enzyme activity, including acidic pH on DNA scaffolds, stabilized hydration layer by DNA phosphate backbones, decreased water activity of nanoconfinement, enrichment of substrate molecules on DNA scaffolds, and substrate channeling . However, many questions remain concerning how DNA confinement modifies the local chemical and physical environments that are critical to enzyme activity and stability.…”

Section: Dna Nanocage‐encapsulated Enzymesmentioning

confidence: 99%

Biomimetic Compartments Scaffolded by Nucleic Acid Nanostructures

Monckton

et al. 2019

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The behaviors of living cells are governed by a series of regulated and confined biochemical reactions. The design and successful construction of synthetic cellular reactors can be useful in a broad range of applications that will bring significant scientific and economic impact. Over the past few decades, DNA self‐assembly has enabled the design and fabrication of sophisticated 1D, 2D, and 3D nanostructures, and is applied to organizing a variety of biomolecular components into prescribed 2D and 3D patterns. In this Concept, the recent and exciting progress in DNA‐scaffolded compartmentalizations and their applications in enzyme encapsulation, lipid membrane assembly, artificial transmembrane nanopores, and smart drug delivery are in focus. Taking advantage of these features promises to deliver breakthroughs toward the attainment of new synthetic and biomimetic reactors.

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Toward Rational Design of High-efficiency Enzyme Cascades

Cited by 183 publications

References 117 publications

Self-assembling all-enzyme hydrogels for biocatalytic flow processes

Self-assembling all-enzyme hydrogels for biocatalytic flow processes

DNA-Scaffolded Proximity Assembly and Confinement of Multienzyme Reactions

Biomimetic Compartments Scaffolded by Nucleic Acid Nanostructures

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