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Enhanced Catalysis from Multienzyme Cascades Assembled on a DNA Origami Triangle

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Standard

Enhanced Catalysis from Multienzyme Cascades Assembled on a DNA Origami Triangle. / Klein, William P.; Thomsen, Rasmus P.; Turner, Kendrick B.; Walper, Scott A.; Vranish, James; Kjems, Jorgen; Ancona, Mario G.; Medintz, Igor L.

I: ACS Nano, Bind 13, Nr. 12, 12.2019, s. 13677-13689.

Publikation: Bidrag til tidsskrift/Konferencebidrag i tidsskrift /Bidrag til avisTidsskriftartikelForskningpeer review

Harvard

Klein, WP, Thomsen, RP, Turner, KB, Walper, SA, Vranish, J, Kjems, J, Ancona, MG & Medintz, IL 2019, 'Enhanced Catalysis from Multienzyme Cascades Assembled on a DNA Origami Triangle', ACS Nano, bind 13, nr. 12, s. 13677-13689. https://doi.org/10.1021/acsnano.9b05746

APA

Klein, W. P., Thomsen, R. P., Turner, K. B., Walper, S. A., Vranish, J., Kjems, J., Ancona, M. G., & Medintz, I. L. (2019). Enhanced Catalysis from Multienzyme Cascades Assembled on a DNA Origami Triangle. ACS Nano, 13(12), 13677-13689. https://doi.org/10.1021/acsnano.9b05746

CBE

Klein WP, Thomsen RP, Turner KB, Walper SA, Vranish J, Kjems J, Ancona MG, Medintz IL. 2019. Enhanced Catalysis from Multienzyme Cascades Assembled on a DNA Origami Triangle. ACS Nano. 13(12):13677-13689. https://doi.org/10.1021/acsnano.9b05746

MLA

Vancouver

Klein WP, Thomsen RP, Turner KB, Walper SA, Vranish J, Kjems J o.a. Enhanced Catalysis from Multienzyme Cascades Assembled on a DNA Origami Triangle. ACS Nano. 2019 dec;13(12):13677-13689. https://doi.org/10.1021/acsnano.9b05746

Author

Klein, William P. ; Thomsen, Rasmus P. ; Turner, Kendrick B. ; Walper, Scott A. ; Vranish, James ; Kjems, Jorgen ; Ancona, Mario G. ; Medintz, Igor L. / Enhanced Catalysis from Multienzyme Cascades Assembled on a DNA Origami Triangle. I: ACS Nano. 2019 ; Bind 13, Nr. 12. s. 13677-13689.

Bibtex

@article{c6985919143a46408d12aab35419c753,
title = "Enhanced Catalysis from Multienzyme Cascades Assembled on a DNA Origami Triangle",
abstract = "Developing reliable methods of constructing cell-free multienzyme biocatalytic systems is a milestone goal of synthetic biology. It would enable overcoming the limitations of current cell-based systems, which suffer from the presence of competing pathways, toxicity, and inefficient access to extracellular reactants and removal of products. DNA nanostructures have been suggested as ideal scaffolds for assembling sequential enzymatic cascades in close enough proximity to potentially allow for exploiting of channeling effects; however, initial demonstrations have provided somewhat contradictory results toward confirming this phenomenon. In this work, a three-enzyme sequential cascade was realized by site-specifically immobilizing DNA-conjugated amylase, maltase, and glucokinase on a self-assembled DNA origami triangle. The kinetics of seven different enzyme configurations were evaluated experimentally and compared to simulations of optimized activity. A 30-fold increase in the pathway's kinetic activity was observed for enzymes assembled to the DNA. Detailed kinetic analysis suggests that this catalytic enhancement originated from increased enzyme stability and a localized DNA surface affinity or hydration layer effect and not from a directed enzyme-to-enzyme channeling mechanism. Nevertheless, the approach used to construct this pathway still shows promise toward improving other more elaborate multienzymatic cascades and could potentially allow for the custom synthesis of complex (bio)molecules that cannot be realized with conventional organic chemistry approaches.",
keywords = "DNA, origami, cell-free, enzyme, cascade, kinetics, channeling, ESCHERICHIA-COLI, BETA-AMYLASE, ENZYME, PURIFICATION, COMPLEXES, NANOSTRUCTURES, PROTEOLYSIS, GLUCOKINASE, EXPRESSION, TRANSPORT",
author = "Klein, {William P.} and Thomsen, {Rasmus P.} and Turner, {Kendrick B.} and Walper, {Scott A.} and James Vranish and Jorgen Kjems and Ancona, {Mario G.} and Medintz, {Igor L.}",
year = "2019",
month = dec,
doi = "10.1021/acsnano.9b05746",
language = "English",
volume = "13",
pages = "13677--13689",
journal = "A C S Nano",
issn = "1936-0851",
publisher = "American Chemical Society",
number = "12",

}

RIS

TY - JOUR

T1 - Enhanced Catalysis from Multienzyme Cascades Assembled on a DNA Origami Triangle

AU - Klein, William P.

AU - Thomsen, Rasmus P.

AU - Turner, Kendrick B.

AU - Walper, Scott A.

AU - Vranish, James

AU - Kjems, Jorgen

AU - Ancona, Mario G.

AU - Medintz, Igor L.

PY - 2019/12

Y1 - 2019/12

N2 - Developing reliable methods of constructing cell-free multienzyme biocatalytic systems is a milestone goal of synthetic biology. It would enable overcoming the limitations of current cell-based systems, which suffer from the presence of competing pathways, toxicity, and inefficient access to extracellular reactants and removal of products. DNA nanostructures have been suggested as ideal scaffolds for assembling sequential enzymatic cascades in close enough proximity to potentially allow for exploiting of channeling effects; however, initial demonstrations have provided somewhat contradictory results toward confirming this phenomenon. In this work, a three-enzyme sequential cascade was realized by site-specifically immobilizing DNA-conjugated amylase, maltase, and glucokinase on a self-assembled DNA origami triangle. The kinetics of seven different enzyme configurations were evaluated experimentally and compared to simulations of optimized activity. A 30-fold increase in the pathway's kinetic activity was observed for enzymes assembled to the DNA. Detailed kinetic analysis suggests that this catalytic enhancement originated from increased enzyme stability and a localized DNA surface affinity or hydration layer effect and not from a directed enzyme-to-enzyme channeling mechanism. Nevertheless, the approach used to construct this pathway still shows promise toward improving other more elaborate multienzymatic cascades and could potentially allow for the custom synthesis of complex (bio)molecules that cannot be realized with conventional organic chemistry approaches.

AB - Developing reliable methods of constructing cell-free multienzyme biocatalytic systems is a milestone goal of synthetic biology. It would enable overcoming the limitations of current cell-based systems, which suffer from the presence of competing pathways, toxicity, and inefficient access to extracellular reactants and removal of products. DNA nanostructures have been suggested as ideal scaffolds for assembling sequential enzymatic cascades in close enough proximity to potentially allow for exploiting of channeling effects; however, initial demonstrations have provided somewhat contradictory results toward confirming this phenomenon. In this work, a three-enzyme sequential cascade was realized by site-specifically immobilizing DNA-conjugated amylase, maltase, and glucokinase on a self-assembled DNA origami triangle. The kinetics of seven different enzyme configurations were evaluated experimentally and compared to simulations of optimized activity. A 30-fold increase in the pathway's kinetic activity was observed for enzymes assembled to the DNA. Detailed kinetic analysis suggests that this catalytic enhancement originated from increased enzyme stability and a localized DNA surface affinity or hydration layer effect and not from a directed enzyme-to-enzyme channeling mechanism. Nevertheless, the approach used to construct this pathway still shows promise toward improving other more elaborate multienzymatic cascades and could potentially allow for the custom synthesis of complex (bio)molecules that cannot be realized with conventional organic chemistry approaches.

KW - DNA

KW - origami

KW - cell-free

KW - enzyme

KW - cascade

KW - kinetics

KW - channeling

KW - ESCHERICHIA-COLI

KW - BETA-AMYLASE

KW - ENZYME

KW - PURIFICATION

KW - COMPLEXES

KW - NANOSTRUCTURES

KW - PROTEOLYSIS

KW - GLUCOKINASE

KW - EXPRESSION

KW - TRANSPORT

U2 - 10.1021/acsnano.9b05746

DO - 10.1021/acsnano.9b05746

M3 - Journal article

VL - 13

SP - 13677

EP - 13689

JO - A C S Nano

JF - A C S Nano

SN - 1936-0851

IS - 12

ER -