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Functional and Molecular Characterization of the Halomicrobium sp. IBSBa Inulosucrase

Fructans are fructose-based (poly)saccharides with inulin and levan being the best-known ones. Thanks to their health-related benefits, inulin-type fructans have been under the focus of scientific and industrial communities, though mostly represented by plant-based inulins, and rarely by microbial o...

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Autores principales: Abaramak, Gülbahar, Porras-Domínguez, Jaime Ricardo, Janse van Rensburg, Henry Christopher, Lescrinier, Eveline, Toksoy Öner, Ebru, Kırtel, Onur, Van den Ende, Wim
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8066391/
https://www.ncbi.nlm.nih.gov/pubmed/33918392
http://dx.doi.org/10.3390/microorganisms9040749
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author Abaramak, Gülbahar
Porras-Domínguez, Jaime Ricardo
Janse van Rensburg, Henry Christopher
Lescrinier, Eveline
Toksoy Öner, Ebru
Kırtel, Onur
Van den Ende, Wim
author_facet Abaramak, Gülbahar
Porras-Domínguez, Jaime Ricardo
Janse van Rensburg, Henry Christopher
Lescrinier, Eveline
Toksoy Öner, Ebru
Kırtel, Onur
Van den Ende, Wim
author_sort Abaramak, Gülbahar
collection PubMed
description Fructans are fructose-based (poly)saccharides with inulin and levan being the best-known ones. Thanks to their health-related benefits, inulin-type fructans have been under the focus of scientific and industrial communities, though mostly represented by plant-based inulins, and rarely by microbial ones. Recently, it was discovered that some extremely halophilic Archaea are also able to synthesize fructans. Here, we describe the first in-depth functional and molecular characterization of an Archaeal inulosucrase from Halomicrobium sp. IBSBa (HmcIsc). The HmcIsc enzyme was recombinantly expressed and purified in Escherichia coli and shown to synthesize inulin as proven by nuclear magnetic resonance (NMR) analysis. In accordance with the halophilic lifestyle of its native host, the enzyme showed maximum activity at very high NaCl concentrations (3.5 M), with specific adaptations for that purpose. Phylogenetic analyses suggested that Archaeal inulosucrases have been acquired from halophilic bacilli through horizontal gene transfer, with a HX(H/F)T motif evolving further into a HXHT motif, together with a unique D residue creating the onset of a specific alternative acceptor binding groove. This work uncovers a novel area in fructan research, highlighting unexplored aspects of life in hypersaline habitats, and raising questions about the general physiological relevance of inulosucrases and their products in nature.
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spelling pubmed-80663912021-04-25 Functional and Molecular Characterization of the Halomicrobium sp. IBSBa Inulosucrase Abaramak, Gülbahar Porras-Domínguez, Jaime Ricardo Janse van Rensburg, Henry Christopher Lescrinier, Eveline Toksoy Öner, Ebru Kırtel, Onur Van den Ende, Wim Microorganisms Article Fructans are fructose-based (poly)saccharides with inulin and levan being the best-known ones. Thanks to their health-related benefits, inulin-type fructans have been under the focus of scientific and industrial communities, though mostly represented by plant-based inulins, and rarely by microbial ones. Recently, it was discovered that some extremely halophilic Archaea are also able to synthesize fructans. Here, we describe the first in-depth functional and molecular characterization of an Archaeal inulosucrase from Halomicrobium sp. IBSBa (HmcIsc). The HmcIsc enzyme was recombinantly expressed and purified in Escherichia coli and shown to synthesize inulin as proven by nuclear magnetic resonance (NMR) analysis. In accordance with the halophilic lifestyle of its native host, the enzyme showed maximum activity at very high NaCl concentrations (3.5 M), with specific adaptations for that purpose. Phylogenetic analyses suggested that Archaeal inulosucrases have been acquired from halophilic bacilli through horizontal gene transfer, with a HX(H/F)T motif evolving further into a HXHT motif, together with a unique D residue creating the onset of a specific alternative acceptor binding groove. This work uncovers a novel area in fructan research, highlighting unexplored aspects of life in hypersaline habitats, and raising questions about the general physiological relevance of inulosucrases and their products in nature. MDPI 2021-04-02 /pmc/articles/PMC8066391/ /pubmed/33918392 http://dx.doi.org/10.3390/microorganisms9040749 Text en © 2021 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Abaramak, Gülbahar
Porras-Domínguez, Jaime Ricardo
Janse van Rensburg, Henry Christopher
Lescrinier, Eveline
Toksoy Öner, Ebru
Kırtel, Onur
Van den Ende, Wim
Functional and Molecular Characterization of the Halomicrobium sp. IBSBa Inulosucrase
title Functional and Molecular Characterization of the Halomicrobium sp. IBSBa Inulosucrase
title_full Functional and Molecular Characterization of the Halomicrobium sp. IBSBa Inulosucrase
title_fullStr Functional and Molecular Characterization of the Halomicrobium sp. IBSBa Inulosucrase
title_full_unstemmed Functional and Molecular Characterization of the Halomicrobium sp. IBSBa Inulosucrase
title_short Functional and Molecular Characterization of the Halomicrobium sp. IBSBa Inulosucrase
title_sort functional and molecular characterization of the halomicrobium sp. ibsba inulosucrase
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8066391/
https://www.ncbi.nlm.nih.gov/pubmed/33918392
http://dx.doi.org/10.3390/microorganisms9040749
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