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Optimized expression of Hfq protein increases Escherichia coli growth
Escherichia coli is a widely used platform for metabolic engineering due to its fast growth and well-established engineering techniques. However, there has been a demand for faster-growing E. coli for higher production of desired substances. Here, to increase the growth of E. coli cells, we optimize...
Autores principales: | , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2021
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7890833/ https://www.ncbi.nlm.nih.gov/pubmed/33602295 http://dx.doi.org/10.1186/s13036-021-00260-x |
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author | VO, Phuong N. L. LEE, Hyang-Mi REN, Jun NA, Dokyun |
author_facet | VO, Phuong N. L. LEE, Hyang-Mi REN, Jun NA, Dokyun |
author_sort | VO, Phuong N. L. |
collection | PubMed |
description | Escherichia coli is a widely used platform for metabolic engineering due to its fast growth and well-established engineering techniques. However, there has been a demand for faster-growing E. coli for higher production of desired substances. Here, to increase the growth of E. coli cells, we optimized the expression level of Hfq protein, which plays an essential role in stress responses. Six variants of the hfq gene with a different ribosome binding site sequence and thereby a different expression level were constructed. When the Hfq expression level was optimized in DH5α, its growth rate was increased by 12.1% and its cell density was also increased by 4.5%. RNA-seq and network analyses revealed the upregulation of stress response genes and metabolic genes, which increases the tolerance against pH changes. When the same strategy was applied to five other E. coli strains (BL21 (DE3), JM109, TOP10, W3110, and MG1655), all their growth rates were increased by 18–94% but not all their densities were increased (− 12 − + 32%). In conclusion, the Hfq expression optimization can increase cell growth rate and probably their cell densities as well. Since the hfq gene is highly conserved across bacterial species, the same strategy could be applied to other bacterial species to construct faster-growing strains. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s13036-021-00260-x. |
format | Online Article Text |
id | pubmed-7890833 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-78908332021-02-22 Optimized expression of Hfq protein increases Escherichia coli growth VO, Phuong N. L. LEE, Hyang-Mi REN, Jun NA, Dokyun J Biol Eng Research Escherichia coli is a widely used platform for metabolic engineering due to its fast growth and well-established engineering techniques. However, there has been a demand for faster-growing E. coli for higher production of desired substances. Here, to increase the growth of E. coli cells, we optimized the expression level of Hfq protein, which plays an essential role in stress responses. Six variants of the hfq gene with a different ribosome binding site sequence and thereby a different expression level were constructed. When the Hfq expression level was optimized in DH5α, its growth rate was increased by 12.1% and its cell density was also increased by 4.5%. RNA-seq and network analyses revealed the upregulation of stress response genes and metabolic genes, which increases the tolerance against pH changes. When the same strategy was applied to five other E. coli strains (BL21 (DE3), JM109, TOP10, W3110, and MG1655), all their growth rates were increased by 18–94% but not all their densities were increased (− 12 − + 32%). In conclusion, the Hfq expression optimization can increase cell growth rate and probably their cell densities as well. Since the hfq gene is highly conserved across bacterial species, the same strategy could be applied to other bacterial species to construct faster-growing strains. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s13036-021-00260-x. BioMed Central 2021-02-18 /pmc/articles/PMC7890833/ /pubmed/33602295 http://dx.doi.org/10.1186/s13036-021-00260-x Text en © The Author(s) 2021 Open AccessThis 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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data. |
spellingShingle | Research VO, Phuong N. L. LEE, Hyang-Mi REN, Jun NA, Dokyun Optimized expression of Hfq protein increases Escherichia coli growth |
title | Optimized expression of Hfq protein increases Escherichia coli growth |
title_full | Optimized expression of Hfq protein increases Escherichia coli growth |
title_fullStr | Optimized expression of Hfq protein increases Escherichia coli growth |
title_full_unstemmed | Optimized expression of Hfq protein increases Escherichia coli growth |
title_short | Optimized expression of Hfq protein increases Escherichia coli growth |
title_sort | optimized expression of hfq protein increases escherichia coli growth |
topic | Research |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7890833/ https://www.ncbi.nlm.nih.gov/pubmed/33602295 http://dx.doi.org/10.1186/s13036-021-00260-x |
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