Cargando…

Improving phloroglucinol tolerance and production in Escherichia coli by GroESL overexpression

BACKGROUND: Phloroglucinol is an important chemical which has been successfully produced by engineered Escherichia coli. However, the toxicity of phloroglucinol can enormously inhibit E. coli cell growth and viability, and the productivity is still too low and not economically feasible for industria...

Descripción completa

Detalles Bibliográficos
Autores principales: Zhang, Rubing, Cao, Yujin, Liu, Wei, Xian, Mo, Liu, Huizhou
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5735909/
https://www.ncbi.nlm.nih.gov/pubmed/29258595
http://dx.doi.org/10.1186/s12934-017-0839-x
_version_ 1783287291246018560
author Zhang, Rubing
Cao, Yujin
Liu, Wei
Xian, Mo
Liu, Huizhou
author_facet Zhang, Rubing
Cao, Yujin
Liu, Wei
Xian, Mo
Liu, Huizhou
author_sort Zhang, Rubing
collection PubMed
description BACKGROUND: Phloroglucinol is an important chemical which has been successfully produced by engineered Escherichia coli. However, the toxicity of phloroglucinol can enormously inhibit E. coli cell growth and viability, and the productivity is still too low and not economically feasible for industrial applications. Therefore, strain tolerance to toxic metabolites remains a key issue during the production of chemicals using biological processes. RESULTS: In the present work, we examined the impact of the native GroESL chaperone system with different overexpression levels on phloroglucinol tolerance and production in E. coli. The groESL gene was cloned into an expression vector, of which expression level was regulated by three different promoters (natural, tac and T7 promoter). Strain tolerance was evaluated employing viable cell counts and phloroglucinol production. In comparison with the control strain, all GroESL overexpressing strains showed good characteristics in cell viability and phloroglucinol synthesis. Strain which overexpressed GroESL under tac promoter was found to show the best tolerance in all of those tested, resulting in a 3.19-fold increase in viable cell numbers compared with control strain of agar-plate culture under the condition of 0.7 g/L phloroglucinol, and a 39.5% increase in phloroglucinol production under fed-batch fermentation. This engineered strain finally accumulated phloroglucinol up to 5.3 g/L in the fed-batch cultivation 10 h after induction, and the productivity was 0.53 g/L/h. To date, the highest phloroglucinol production was achieved in this work compared with the previous reports, which is promising to make the bioprocess feasible from the economical point. CONCLUSIONS: The data show that appropriate expression level of GroESL plays a critical role in improving phloroglucinol tolerance and production in E. coli, and maybe involve in controlling some aspects of the stress response system through upregulation of GroESL. GroESL overexpression is therefore a feasible and efficient approach for improvement of E. coli tolerance. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s12934-017-0839-x) contains supplementary material, which is available to authorized users.
format Online
Article
Text
id pubmed-5735909
institution National Center for Biotechnology Information
language English
publishDate 2017
publisher BioMed Central
record_format MEDLINE/PubMed
spelling pubmed-57359092017-12-21 Improving phloroglucinol tolerance and production in Escherichia coli by GroESL overexpression Zhang, Rubing Cao, Yujin Liu, Wei Xian, Mo Liu, Huizhou Microb Cell Fact Research BACKGROUND: Phloroglucinol is an important chemical which has been successfully produced by engineered Escherichia coli. However, the toxicity of phloroglucinol can enormously inhibit E. coli cell growth and viability, and the productivity is still too low and not economically feasible for industrial applications. Therefore, strain tolerance to toxic metabolites remains a key issue during the production of chemicals using biological processes. RESULTS: In the present work, we examined the impact of the native GroESL chaperone system with different overexpression levels on phloroglucinol tolerance and production in E. coli. The groESL gene was cloned into an expression vector, of which expression level was regulated by three different promoters (natural, tac and T7 promoter). Strain tolerance was evaluated employing viable cell counts and phloroglucinol production. In comparison with the control strain, all GroESL overexpressing strains showed good characteristics in cell viability and phloroglucinol synthesis. Strain which overexpressed GroESL under tac promoter was found to show the best tolerance in all of those tested, resulting in a 3.19-fold increase in viable cell numbers compared with control strain of agar-plate culture under the condition of 0.7 g/L phloroglucinol, and a 39.5% increase in phloroglucinol production under fed-batch fermentation. This engineered strain finally accumulated phloroglucinol up to 5.3 g/L in the fed-batch cultivation 10 h after induction, and the productivity was 0.53 g/L/h. To date, the highest phloroglucinol production was achieved in this work compared with the previous reports, which is promising to make the bioprocess feasible from the economical point. CONCLUSIONS: The data show that appropriate expression level of GroESL plays a critical role in improving phloroglucinol tolerance and production in E. coli, and maybe involve in controlling some aspects of the stress response system through upregulation of GroESL. GroESL overexpression is therefore a feasible and efficient approach for improvement of E. coli tolerance. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s12934-017-0839-x) contains supplementary material, which is available to authorized users. BioMed Central 2017-12-19 /pmc/articles/PMC5735909/ /pubmed/29258595 http://dx.doi.org/10.1186/s12934-017-0839-x Text en © The Author(s) 2017 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, Rubing
Cao, Yujin
Liu, Wei
Xian, Mo
Liu, Huizhou
Improving phloroglucinol tolerance and production in Escherichia coli by GroESL overexpression
title Improving phloroglucinol tolerance and production in Escherichia coli by GroESL overexpression
title_full Improving phloroglucinol tolerance and production in Escherichia coli by GroESL overexpression
title_fullStr Improving phloroglucinol tolerance and production in Escherichia coli by GroESL overexpression
title_full_unstemmed Improving phloroglucinol tolerance and production in Escherichia coli by GroESL overexpression
title_short Improving phloroglucinol tolerance and production in Escherichia coli by GroESL overexpression
title_sort improving phloroglucinol tolerance and production in escherichia coli by groesl overexpression
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5735909/
https://www.ncbi.nlm.nih.gov/pubmed/29258595
http://dx.doi.org/10.1186/s12934-017-0839-x
work_keys_str_mv AT zhangrubing improvingphloroglucinoltoleranceandproductioninescherichiacolibygroesloverexpression
AT caoyujin improvingphloroglucinoltoleranceandproductioninescherichiacolibygroesloverexpression
AT liuwei improvingphloroglucinoltoleranceandproductioninescherichiacolibygroesloverexpression
AT xianmo improvingphloroglucinoltoleranceandproductioninescherichiacolibygroesloverexpression
AT liuhuizhou improvingphloroglucinoltoleranceandproductioninescherichiacolibygroesloverexpression