Harvesting of Prebiotic Fructooligosaccharides by Nonbeneficial Human Gut Bacteria

ABSTRACT Prebiotic oligosaccharides, such as fructooligosaccharides, are increasingly being used to modulate the composition and activity of the gut microbiota. However, carbohydrate utilization analyses and metagenomic studies recently revealed the ability of deleterious and uncultured human gut ba...

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Autores principales:	Zhi Wang, Alexandra S. Tauzin, Elisabeth Laville, Pietro Tedesco, Fabien Létisse, Nicolas Terrapon, Pascale Lepercq, Myriam Mercade, Gabrielle Potocki-Veronese
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2020
Materias:	Dorea chronic gut diseases fructooligosaccharides microbiome phosphotransferase system Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/256f44d6bf444cd98325e1fccfd71fb5
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spelling	oai:doaj.org-article:256f44d6bf444cd98325e1fccfd71fb52021-11-15T15:27:53ZHarvesting of Prebiotic Fructooligosaccharides by Nonbeneficial Human Gut Bacteria10.1128/mSphere.00771-192379-5042https://doaj.org/article/256f44d6bf444cd98325e1fccfd71fb52020-02-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mSphere.00771-19https://doaj.org/toc/2379-5042ABSTRACT Prebiotic oligosaccharides, such as fructooligosaccharides, are increasingly being used to modulate the composition and activity of the gut microbiota. However, carbohydrate utilization analyses and metagenomic studies recently revealed the ability of deleterious and uncultured human gut bacterial species to metabolize these functional foods. Moreover, because of the difficulties of functionally profiling transmembrane proteins, only a few prebiotic transporters have been biochemically characterized to date, while carbohydrate binding and transport are the first and thus crucial steps in their metabolization. Here, we describe the molecular mechanism of a phosphotransferase system, highlighted as a dietary and pathology biomarker in the human gut microbiome. This transporter is encoded by a metagenomic locus that is highly conserved in several human gut Firmicutes, including Dorea species. We developed a generic strategy to deeply analyze, in vitro and in cellulo, the specificity and functionality of recombinant transporters in Escherichia coli, combining carbohydrate utilization locus and host genome engineering and quantification of the binding, transport, and growth rates with analysis of phosphorylated carbohydrates by mass spectrometry. We demonstrated that the Dorea fructooligosaccharide transporter is specific for kestose, whether for binding, transport, or phosphorylation. This constitutes the biochemical proof of effective phosphorylation of glycosides with a degree of polymerization of more than 2, extending the known functional diversity of phosphotransferase systems. Based on these new findings, we revisited the classification of these carbohydrate transporters. IMPORTANCE Prebiotics are increasingly used as food supplements, especially in infant formulas, to modify the functioning and composition of the microbiota. However, little is currently known about the mechanisms of prebiotic recognition and transport by gut bacteria, while these steps are crucial in their metabolism. In this study, we established a new strategy to profile the specificity of oligosaccharide transporters, combining microbiomics, genetic locus and strain engineering, and state-of-the art metabolomics. We revisited the transporter classification database and proposed a new way to classify these membrane proteins based on their structural and mechanistic similarities. Based on these developments, we identified and characterized, at the molecular level, a fructooligosaccharide transporting phosphotransferase system, which constitutes a biomarker of diet and gut pathology. The deciphering of this prebiotic metabolization mechanism by a nonbeneficial bacterium highlights the controversial use of prebiotics, especially in the context of chronic gut diseases.Zhi WangAlexandra S. TauzinElisabeth LavillePietro TedescoFabien LétisseNicolas TerraponPascale LepercqMyriam MercadeGabrielle Potocki-VeroneseAmerican Society for MicrobiologyarticleDoreachronic gut diseasesfructooligosaccharidesmicrobiomephosphotransferase systemMicrobiologyQR1-502ENmSphere, Vol 5, Iss 1 (2020)
institution	DOAJ
collection	DOAJ
language	EN
topic	Dorea chronic gut diseases fructooligosaccharides microbiome phosphotransferase system Microbiology QR1-502
spellingShingle	Dorea chronic gut diseases fructooligosaccharides microbiome phosphotransferase system Microbiology QR1-502 Zhi Wang Alexandra S. Tauzin Elisabeth Laville Pietro Tedesco Fabien Létisse Nicolas Terrapon Pascale Lepercq Myriam Mercade Gabrielle Potocki-Veronese Harvesting of Prebiotic Fructooligosaccharides by Nonbeneficial Human Gut Bacteria
description	ABSTRACT Prebiotic oligosaccharides, such as fructooligosaccharides, are increasingly being used to modulate the composition and activity of the gut microbiota. However, carbohydrate utilization analyses and metagenomic studies recently revealed the ability of deleterious and uncultured human gut bacterial species to metabolize these functional foods. Moreover, because of the difficulties of functionally profiling transmembrane proteins, only a few prebiotic transporters have been biochemically characterized to date, while carbohydrate binding and transport are the first and thus crucial steps in their metabolization. Here, we describe the molecular mechanism of a phosphotransferase system, highlighted as a dietary and pathology biomarker in the human gut microbiome. This transporter is encoded by a metagenomic locus that is highly conserved in several human gut Firmicutes, including Dorea species. We developed a generic strategy to deeply analyze, in vitro and in cellulo, the specificity and functionality of recombinant transporters in Escherichia coli, combining carbohydrate utilization locus and host genome engineering and quantification of the binding, transport, and growth rates with analysis of phosphorylated carbohydrates by mass spectrometry. We demonstrated that the Dorea fructooligosaccharide transporter is specific for kestose, whether for binding, transport, or phosphorylation. This constitutes the biochemical proof of effective phosphorylation of glycosides with a degree of polymerization of more than 2, extending the known functional diversity of phosphotransferase systems. Based on these new findings, we revisited the classification of these carbohydrate transporters. IMPORTANCE Prebiotics are increasingly used as food supplements, especially in infant formulas, to modify the functioning and composition of the microbiota. However, little is currently known about the mechanisms of prebiotic recognition and transport by gut bacteria, while these steps are crucial in their metabolism. In this study, we established a new strategy to profile the specificity of oligosaccharide transporters, combining microbiomics, genetic locus and strain engineering, and state-of-the art metabolomics. We revisited the transporter classification database and proposed a new way to classify these membrane proteins based on their structural and mechanistic similarities. Based on these developments, we identified and characterized, at the molecular level, a fructooligosaccharide transporting phosphotransferase system, which constitutes a biomarker of diet and gut pathology. The deciphering of this prebiotic metabolization mechanism by a nonbeneficial bacterium highlights the controversial use of prebiotics, especially in the context of chronic gut diseases.
format	article
author	Zhi Wang Alexandra S. Tauzin Elisabeth Laville Pietro Tedesco Fabien Létisse Nicolas Terrapon Pascale Lepercq Myriam Mercade Gabrielle Potocki-Veronese
author_facet	Zhi Wang Alexandra S. Tauzin Elisabeth Laville Pietro Tedesco Fabien Létisse Nicolas Terrapon Pascale Lepercq Myriam Mercade Gabrielle Potocki-Veronese
author_sort	Zhi Wang
title	Harvesting of Prebiotic Fructooligosaccharides by Nonbeneficial Human Gut Bacteria
title_short	Harvesting of Prebiotic Fructooligosaccharides by Nonbeneficial Human Gut Bacteria
title_full	Harvesting of Prebiotic Fructooligosaccharides by Nonbeneficial Human Gut Bacteria
title_fullStr	Harvesting of Prebiotic Fructooligosaccharides by Nonbeneficial Human Gut Bacteria
title_full_unstemmed	Harvesting of Prebiotic Fructooligosaccharides by Nonbeneficial Human Gut Bacteria
title_sort	harvesting of prebiotic fructooligosaccharides by nonbeneficial human gut bacteria
publisher	American Society for Microbiology
publishDate	2020
url	https://doaj.org/article/256f44d6bf444cd98325e1fccfd71fb5
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Harvesting of Prebiotic Fructooligosaccharides by Nonbeneficial Human Gut Bacteria

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