Genome-scale target identification in Escherichia coli for high-titer production of free fatty acids

Lixia Fang, Jie Fan, Shulei Luo, Yaru Chen, Congya Wang, Yingxiu Cao () and Hao Song ()
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Lixia Fang: Tianjin University
Jie Fan: Tianjin University
Shulei Luo: Tianjin University
Yaru Chen: Tianjin University
Congya Wang: Tianjin University
Yingxiu Cao: Tianjin University
Hao Song: Tianjin University

Nature Communications, 2021, vol. 12, issue 1, 1-13

Abstract: Abstract To construct a superior microbial cell factory for chemical synthesis, a major challenge is to fully exploit cellular potential by identifying and engineering beneficial gene targets in sophisticated metabolic networks. Here, we take advantage of CRISPR interference (CRISPRi) and omics analyses to systematically identify beneficial genes that can be engineered to promote free fatty acids (FFAs) production in Escherichia coli. CRISPRi-mediated genetic perturbation enables the identification of 30 beneficial genes from 108 targets related to FFA metabolism. Then, omics analyses of the FFAs-overproducing strains and a control strain enable the identification of another 26 beneficial genes that are seemingly irrelevant to FFA metabolism. Combinatorial perturbation of four beneficial genes involving cellular stress responses results in a recombinant strain ihfAL−-aidB+-ryfAM−-gadAH−, producing 30.0 g L−1 FFAs in fed-batch fermentation, the maximum titer in E. coli reported to date. Our findings are of help in rewiring cellular metabolism and interwoven intracellular processes to facilitate high-titer production of biochemicals.

Date: 2021
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DOI: 10.1038/s41467-021-25243-w

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