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Significantly improved solvent tolerance of Escherichia coli by global transcription machinery engineering

BACKGROUND: Escherichia coli has emerged as a promising platform microorganism to produce biofuels and fine chemicals of industrial interests. Certain obstacles however remain to be overcome, among which organic-solvent tolerance is a crucial one. RESULTS: We used global transcription machinery engi...

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Autores principales: Zhang, Fa, Qian, Xiaohong, Si, Haiming, Xu, Guochao, Han, Ruizhi, Ni, Ye
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2015
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4635540/
https://www.ncbi.nlm.nih.gov/pubmed/26542360
http://dx.doi.org/10.1186/s12934-015-0368-4
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author Zhang, Fa
Qian, Xiaohong
Si, Haiming
Xu, Guochao
Han, Ruizhi
Ni, Ye
author_facet Zhang, Fa
Qian, Xiaohong
Si, Haiming
Xu, Guochao
Han, Ruizhi
Ni, Ye
author_sort Zhang, Fa
collection PubMed
description BACKGROUND: Escherichia coli has emerged as a promising platform microorganism to produce biofuels and fine chemicals of industrial interests. Certain obstacles however remain to be overcome, among which organic-solvent tolerance is a crucial one. RESULTS: We used global transcription machinery engineering (gTME) to improve the organic-solvent tolerance (OST) of E. coli JM109. A mutant library of σ(70) encoded by rpoD was screened under cyclohexane pressure. E. coli JM109 strain harboring σ(70) mutant C9 was identified with capability of tolerating 69 % cyclohexane. The rpoD mutant contains three amino-acid substitutes and a stop-codon mutation, resulting a truncated sequence containing regions σ(1.1) and σ(1.2). Total protein difference produced by E. coli JM109 strain harboring C9 was examined with 2D-PAGE, and 204 high-abundant proteins showed over twofold variation under different solvent stress. CONCLUSIONS: Our results show that several genes (gapA, sdhB, pepB and dppA) play critical roles in enhanced solvent tolerance of E. coli, mainly involving in maintaining higher intracellular energy level under solvent stress. Global transcription machinery engineering is therefore a feasible and efficient approach for engineering strain with enhanced OST-phenotype. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s12934-015-0368-4) contains supplementary material, which is available to authorized users.
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spelling pubmed-46355402015-11-07 Significantly improved solvent tolerance of Escherichia coli by global transcription machinery engineering Zhang, Fa Qian, Xiaohong Si, Haiming Xu, Guochao Han, Ruizhi Ni, Ye Microb Cell Fact Research BACKGROUND: Escherichia coli has emerged as a promising platform microorganism to produce biofuels and fine chemicals of industrial interests. Certain obstacles however remain to be overcome, among which organic-solvent tolerance is a crucial one. RESULTS: We used global transcription machinery engineering (gTME) to improve the organic-solvent tolerance (OST) of E. coli JM109. A mutant library of σ(70) encoded by rpoD was screened under cyclohexane pressure. E. coli JM109 strain harboring σ(70) mutant C9 was identified with capability of tolerating 69 % cyclohexane. The rpoD mutant contains three amino-acid substitutes and a stop-codon mutation, resulting a truncated sequence containing regions σ(1.1) and σ(1.2). Total protein difference produced by E. coli JM109 strain harboring C9 was examined with 2D-PAGE, and 204 high-abundant proteins showed over twofold variation under different solvent stress. CONCLUSIONS: Our results show that several genes (gapA, sdhB, pepB and dppA) play critical roles in enhanced solvent tolerance of E. coli, mainly involving in maintaining higher intracellular energy level under solvent stress. Global transcription machinery engineering is therefore a feasible and efficient approach for engineering strain with enhanced OST-phenotype. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s12934-015-0368-4) contains supplementary material, which is available to authorized users. BioMed Central 2015-11-05 /pmc/articles/PMC4635540/ /pubmed/26542360 http://dx.doi.org/10.1186/s12934-015-0368-4 Text en © Zhang et al. 2015 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided 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 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.
spellingShingle Research
Zhang, Fa
Qian, Xiaohong
Si, Haiming
Xu, Guochao
Han, Ruizhi
Ni, Ye
Significantly improved solvent tolerance of Escherichia coli by global transcription machinery engineering
title Significantly improved solvent tolerance of Escherichia coli by global transcription machinery engineering
title_full Significantly improved solvent tolerance of Escherichia coli by global transcription machinery engineering
title_fullStr Significantly improved solvent tolerance of Escherichia coli by global transcription machinery engineering
title_full_unstemmed Significantly improved solvent tolerance of Escherichia coli by global transcription machinery engineering
title_short Significantly improved solvent tolerance of Escherichia coli by global transcription machinery engineering
title_sort significantly improved solvent tolerance of escherichia coli by global transcription machinery engineering
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4635540/
https://www.ncbi.nlm.nih.gov/pubmed/26542360
http://dx.doi.org/10.1186/s12934-015-0368-4
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