AI Article Synopsis
- Researchers used CRISPR interference and omics analyses to find and engineer genes that enhance free fatty acid production in E. coli.
- They identified 30 beneficial genes through genetic perturbations and discovered an additional 26 genes unrelated to fatty acid metabolism by analyzing FFAs-overproducing strains.
- By combining changes in four specific genes related to cellular stress responses, they created a new strain that achieved a record-high free fatty acid production of 30.0 g/L in fermentation.
Article 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 ihfA-aidB-ryfA-gadA, producing 30.0 g L 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.
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|---|---|
| http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8371096 | PMC |
| http://dx.doi.org/10.1038/s41467-021-25243-w | DOI Listing |
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