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An auto-inducible phosphate-controlled expression system of Bacillus licheniformis

BACKGROUND: A promoter that drives high-level, long-term expression of the target gene under substrate limited growth conditions in the absence of an artificial inducer would facilitate the efficient production of heterologous proteins at low cost. A novel phosphate-regulated expression system was c...

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Autores principales: Trung, Nguyen Thanh, Hung, Nguyen Minh, Thuan, Nguyen Huy, Canh, Nguyen Xuan, Schweder, Thomas, Jürgen, Britta
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6327384/
https://www.ncbi.nlm.nih.gov/pubmed/30626366
http://dx.doi.org/10.1186/s12896-018-0490-6
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author Trung, Nguyen Thanh
Hung, Nguyen Minh
Thuan, Nguyen Huy
Canh, Nguyen Xuan
Schweder, Thomas
Jürgen, Britta
author_facet Trung, Nguyen Thanh
Hung, Nguyen Minh
Thuan, Nguyen Huy
Canh, Nguyen Xuan
Schweder, Thomas
Jürgen, Britta
author_sort Trung, Nguyen Thanh
collection PubMed
description BACKGROUND: A promoter that drives high-level, long-term expression of the target gene under substrate limited growth conditions in the absence of an artificial inducer would facilitate the efficient production of heterologous proteins at low cost. A novel phosphate-regulated expression system was constructed using the promoter of the phytase encoding gene phyL from Bacillus licheniformis for the overexpression of proteins in this industrially relevant host. RESULTS: It is shown that the phyL promoter enables a strong overexpression of the heterologous genes amyE and xynA in B. licheniformis when cells were subjected to phosphate limitation. Whether B. licheniformis can use phytate as an alternative phosphate source and how this substrate influences the PphyL controlled gene expression under growth conditions with limited inorganic phosphate concentrations were also investigated. It is shown that B. licheniformis cells are able to use sodium phytate as alternative phosphate source. The addition of small amounts of sodium phytate (≤ 5 mM) to the growth medium resulted in a strong induction and overexpression of both model genes in B. licheniformis cells under phosphate limited growth conditions. CONCLUSIONS: The PphyL controlled expression of the investigated heterologous genes in B. licheniformis is strongly auto-induced under phosphate limited conditions. The proposed PphyL expression system enables an overexpression of target genes in B. licheniformis under growth conditions, which can be easily performed in a fed-batch fermentation process. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s12896-018-0490-6) contains supplementary material, which is available to authorized users.
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spelling pubmed-63273842019-01-15 An auto-inducible phosphate-controlled expression system of Bacillus licheniformis Trung, Nguyen Thanh Hung, Nguyen Minh Thuan, Nguyen Huy Canh, Nguyen Xuan Schweder, Thomas Jürgen, Britta BMC Biotechnol Research Article BACKGROUND: A promoter that drives high-level, long-term expression of the target gene under substrate limited growth conditions in the absence of an artificial inducer would facilitate the efficient production of heterologous proteins at low cost. A novel phosphate-regulated expression system was constructed using the promoter of the phytase encoding gene phyL from Bacillus licheniformis for the overexpression of proteins in this industrially relevant host. RESULTS: It is shown that the phyL promoter enables a strong overexpression of the heterologous genes amyE and xynA in B. licheniformis when cells were subjected to phosphate limitation. Whether B. licheniformis can use phytate as an alternative phosphate source and how this substrate influences the PphyL controlled gene expression under growth conditions with limited inorganic phosphate concentrations were also investigated. It is shown that B. licheniformis cells are able to use sodium phytate as alternative phosphate source. The addition of small amounts of sodium phytate (≤ 5 mM) to the growth medium resulted in a strong induction and overexpression of both model genes in B. licheniformis cells under phosphate limited growth conditions. CONCLUSIONS: The PphyL controlled expression of the investigated heterologous genes in B. licheniformis is strongly auto-induced under phosphate limited conditions. The proposed PphyL expression system enables an overexpression of target genes in B. licheniformis under growth conditions, which can be easily performed in a fed-batch fermentation process. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s12896-018-0490-6) contains supplementary material, which is available to authorized users. BioMed Central 2019-01-09 /pmc/articles/PMC6327384/ /pubmed/30626366 http://dx.doi.org/10.1186/s12896-018-0490-6 Text en © The Author(s). 2019 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 Article
Trung, Nguyen Thanh
Hung, Nguyen Minh
Thuan, Nguyen Huy
Canh, Nguyen Xuan
Schweder, Thomas
Jürgen, Britta
An auto-inducible phosphate-controlled expression system of Bacillus licheniformis
title An auto-inducible phosphate-controlled expression system of Bacillus licheniformis
title_full An auto-inducible phosphate-controlled expression system of Bacillus licheniformis
title_fullStr An auto-inducible phosphate-controlled expression system of Bacillus licheniformis
title_full_unstemmed An auto-inducible phosphate-controlled expression system of Bacillus licheniformis
title_short An auto-inducible phosphate-controlled expression system of Bacillus licheniformis
title_sort auto-inducible phosphate-controlled expression system of bacillus licheniformis
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6327384/
https://www.ncbi.nlm.nih.gov/pubmed/30626366
http://dx.doi.org/10.1186/s12896-018-0490-6
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