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Using fluorescent promoter-reporters to study sugar utilization control in Bifidobacterium longum NCC 2705
Bifidobacteria are amongst the first bacteria to colonize the human gastro-intestinal system and have been proposed to play a crucial role in the development of the infant gut since their absence is correlated to the development of diseases later in life. Bifidobacteria have the capacity to metaboli...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group UK
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9213400/ https://www.ncbi.nlm.nih.gov/pubmed/35729224 http://dx.doi.org/10.1038/s41598-022-14638-4 |
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author | Duboux, S. Muller, J. A. De Franceschi, F. Mercenier, A. Kleerebezem, M. |
author_facet | Duboux, S. Muller, J. A. De Franceschi, F. Mercenier, A. Kleerebezem, M. |
author_sort | Duboux, S. |
collection | PubMed |
description | Bifidobacteria are amongst the first bacteria to colonize the human gastro-intestinal system and have been proposed to play a crucial role in the development of the infant gut since their absence is correlated to the development of diseases later in life. Bifidobacteria have the capacity to metabolize a diverse range of (complex) carbohydrates, reflecting their adaptation to the lower gastro-intestinal tract. Detailed understanding of carbohydrate metabolism regulation in this genus is of prime importance and availability of additional genetic tools easing such studies would be beneficial. To develop a fluorescent protein-based reporter system that can be used in B. longum NCC 2705, we first selected the most promising fluorescent protein out of the seven we tested (i.e., mCherry). This reporter protein was then used to study the carbohydrate mediated activation of P(Bl1518) and P(Bl1694), two promoters respectively predicted to be controlled by the transcriptional factors AraQ and AraU, previously suggested to regulate arabinose utilization and proposed to also act as global transcriptional regulators in bifidobacteria. We confirmed that in B. longum NCC 2705 the AraQ controlled promoter (P(Bl1518)) is induced strongly by arabinose and established that the AraU controlled promoter (P(Bl1694)) was mostly induced by the hexoses galactose and fructose. Combining the mCherry reporter system with flow cytometry, we established that NCC 2705 is able to co-metabolize arabinose and glucose while galactose was only consumed after glucose exhaustion, thus illustrating the complexity of different carbohydrate consumption patterns and their specific regulation in this strain. |
format | Online Article Text |
id | pubmed-9213400 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | Nature Publishing Group UK |
record_format | MEDLINE/PubMed |
spelling | pubmed-92134002022-06-23 Using fluorescent promoter-reporters to study sugar utilization control in Bifidobacterium longum NCC 2705 Duboux, S. Muller, J. A. De Franceschi, F. Mercenier, A. Kleerebezem, M. Sci Rep Article Bifidobacteria are amongst the first bacteria to colonize the human gastro-intestinal system and have been proposed to play a crucial role in the development of the infant gut since their absence is correlated to the development of diseases later in life. Bifidobacteria have the capacity to metabolize a diverse range of (complex) carbohydrates, reflecting their adaptation to the lower gastro-intestinal tract. Detailed understanding of carbohydrate metabolism regulation in this genus is of prime importance and availability of additional genetic tools easing such studies would be beneficial. To develop a fluorescent protein-based reporter system that can be used in B. longum NCC 2705, we first selected the most promising fluorescent protein out of the seven we tested (i.e., mCherry). This reporter protein was then used to study the carbohydrate mediated activation of P(Bl1518) and P(Bl1694), two promoters respectively predicted to be controlled by the transcriptional factors AraQ and AraU, previously suggested to regulate arabinose utilization and proposed to also act as global transcriptional regulators in bifidobacteria. We confirmed that in B. longum NCC 2705 the AraQ controlled promoter (P(Bl1518)) is induced strongly by arabinose and established that the AraU controlled promoter (P(Bl1694)) was mostly induced by the hexoses galactose and fructose. Combining the mCherry reporter system with flow cytometry, we established that NCC 2705 is able to co-metabolize arabinose and glucose while galactose was only consumed after glucose exhaustion, thus illustrating the complexity of different carbohydrate consumption patterns and their specific regulation in this strain. Nature Publishing Group UK 2022-06-21 /pmc/articles/PMC9213400/ /pubmed/35729224 http://dx.doi.org/10.1038/s41598-022-14638-4 Text en © The Author(s) 2022 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 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/ (https://creativecommons.org/licenses/by/4.0/) . |
spellingShingle | Article Duboux, S. Muller, J. A. De Franceschi, F. Mercenier, A. Kleerebezem, M. Using fluorescent promoter-reporters to study sugar utilization control in Bifidobacterium longum NCC 2705 |
title | Using fluorescent promoter-reporters to study sugar utilization control in Bifidobacterium longum NCC 2705 |
title_full | Using fluorescent promoter-reporters to study sugar utilization control in Bifidobacterium longum NCC 2705 |
title_fullStr | Using fluorescent promoter-reporters to study sugar utilization control in Bifidobacterium longum NCC 2705 |
title_full_unstemmed | Using fluorescent promoter-reporters to study sugar utilization control in Bifidobacterium longum NCC 2705 |
title_short | Using fluorescent promoter-reporters to study sugar utilization control in Bifidobacterium longum NCC 2705 |
title_sort | using fluorescent promoter-reporters to study sugar utilization control in bifidobacterium longum ncc 2705 |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9213400/ https://www.ncbi.nlm.nih.gov/pubmed/35729224 http://dx.doi.org/10.1038/s41598-022-14638-4 |
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