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Modular enzyme assembly for enhanced cascade biocatalysis and metabolic flux

Wei Kang, Tian Ma, Min Liu, Jiale Qu, Zhenjun Liu, Huawei Zhang, Bin Shi, Shuai Fu, Juncai Ma, Louis Tung Faat Lai, Sicong He, Jianan Qu, Shannon Wing-Ngor Au, Byung Ho Kang, Wilson Chun Yu Lau, Zixin Deng, Jiang Xia () and Tiangang Liu ()
Additional contact information
Wei Kang: The Chinese University of Hong Kong
Tian Ma: Wuhan University
Min Liu: The Chinese University of Hong Kong
Jiale Qu: The Chinese University of Hong Kong
Zhenjun Liu: The Chinese University of Hong Kong
Huawei Zhang: The Chinese University of Hong Kong
Bin Shi: Wuhan University
Shuai Fu: J1 Biotech Co., Ltd.
Juncai Ma: The Chinese University of Hong Kong
Louis Tung Faat Lai: The Chinese University of Hong Kong
Sicong He: Hong Kong University of Science and Technology
Jianan Qu: Hong Kong University of Science and Technology
Shannon Wing-Ngor Au: The Chinese University of Hong Kong
Byung Ho Kang: The Chinese University of Hong Kong
Wilson Chun Yu Lau: The Chinese University of Hong Kong
Zixin Deng: Wuhan University
Jiang Xia: The Chinese University of Hong Kong
Tiangang Liu: Wuhan University

Nature Communications, 2019, vol. 10, issue 1, 1-11

Abstract: Abstract Enzymatic reactions in living cells are highly dynamic but simultaneously tightly regulated. Enzyme engineers seek to construct multienzyme complexes to prevent intermediate diffusion, to improve product yield, and to control the flux of metabolites. Here we choose a pair of short peptide tags (RIAD and RIDD) to create scaffold-free enzyme assemblies to achieve these goals. In vitro, assembling enzymes in the menaquinone biosynthetic pathway through RIAD–RIDD interaction yields protein nanoparticles with varying stoichiometries, sizes, geometries, and catalytic efficiency. In Escherichia coli, assembling the last enzyme of the upstream mevalonate pathway with the first enzyme of the downstream carotenoid pathway leads to the formation of a pathway node, which increases carotenoid production by 5.7 folds. The same strategy results in a 58% increase in lycopene production in engineered Saccharomyces cerevisiae. This work presents a simple strategy to impose metabolic control in biosynthetic microbe factories.

Date: 2019
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DOI: 10.1038/s41467-019-12247-w

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