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Genome-scale target identification in Escherichia coli for high-titer production of free fatty acids
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 s...
Autores principales: | , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group UK
2021
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8371096/ https://www.ncbi.nlm.nih.gov/pubmed/34404790 http://dx.doi.org/10.1038/s41467-021-25243-w |
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author | Fang, Lixia Fan, Jie Luo, Shulei Chen, Yaru Wang, Congya Cao, Yingxiu Song, Hao |
author_facet | Fang, Lixia Fan, Jie Luo, Shulei Chen, Yaru Wang, Congya Cao, Yingxiu Song, Hao |
author_sort | Fang, Lixia |
collection | PubMed |
description | 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(L−)-aidB(+)-ryfA(M−)-gadA(H−), 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. |
format | Online Article Text |
id | pubmed-8371096 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | Nature Publishing Group UK |
record_format | MEDLINE/PubMed |
spelling | pubmed-83710962021-09-02 Genome-scale target identification in Escherichia coli for high-titer production of free fatty acids Fang, Lixia Fan, Jie Luo, Shulei Chen, Yaru Wang, Congya Cao, Yingxiu Song, Hao Nat Commun Article 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(L−)-aidB(+)-ryfA(M−)-gadA(H−), 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. Nature Publishing Group UK 2021-08-17 /pmc/articles/PMC8371096/ /pubmed/34404790 http://dx.doi.org/10.1038/s41467-021-25243-w Text en © The Author(s) 2021 https://creativecommons.org/licenses/by/4.0/Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) . |
spellingShingle | Article Fang, Lixia Fan, Jie Luo, Shulei Chen, Yaru Wang, Congya Cao, Yingxiu Song, Hao Genome-scale target identification in Escherichia coli for high-titer production of free fatty acids |
title | Genome-scale target identification in Escherichia coli for high-titer production of free fatty acids |
title_full | Genome-scale target identification in Escherichia coli for high-titer production of free fatty acids |
title_fullStr | Genome-scale target identification in Escherichia coli for high-titer production of free fatty acids |
title_full_unstemmed | Genome-scale target identification in Escherichia coli for high-titer production of free fatty acids |
title_short | Genome-scale target identification in Escherichia coli for high-titer production of free fatty acids |
title_sort | genome-scale target identification in escherichia coli for high-titer production of free fatty acids |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8371096/ https://www.ncbi.nlm.nih.gov/pubmed/34404790 http://dx.doi.org/10.1038/s41467-021-25243-w |
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