Author
Listed:
- Lixia Fang
(Tianjin University
Tianjin University)
- Jie Fan
(Tianjin University
Tianjin University)
- Shulei Luo
(Tianjin University
Tianjin University)
- Yaru Chen
(Tianjin University
Tianjin University)
- Congya Wang
(Tianjin University
Tianjin University)
- Yingxiu Cao
(Tianjin University
Tianjin University)
- Hao Song
(Tianjin University
Tianjin University)
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.
Suggested Citation
Lixia Fang & Jie Fan & Shulei Luo & Yaru Chen & Congya Wang & Yingxiu Cao & Hao Song, 2021.
"Genome-scale target identification in Escherichia coli for high-titer production of free fatty acids,"
Nature Communications, Nature, vol. 12(1), pages 1-13, December.
Handle:
RePEc:nat:natcom:v:12:y:2021:i:1:d:10.1038_s41467-021-25243-w
DOI: 10.1038/s41467-021-25243-w
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