Dataset Information

Functional and Molecular Characterization of the Halomicrobium sp. IBSBa Inulosucrase.

ABSTRACT: 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.

SUBMITTER: Abaramak G

PROVIDER: S-EPMC8066391 | biostudies-literature | 2021 Apr

REPOSITORIES: biostudies-literature

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Functional and Molecular Characterization of the <i>Halomicrobium</i> sp. IBSBa Inulosucrase.

Abaramak Gülbahar G Porras-Domínguez Jaime Ricardo JR Janse van Rensburg Henry Christopher HC Lescrinier Eveline E Toksoy Öner Ebru E Kırtel Onur O Van den Ende Wim W

Microorganisms 20210402 4

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 Archa ...[more]

PMID: 33918392

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Project description:IntroductionBlastocystis sp. is a protozoan that colonizes the gastrointestinal tract of humans and many animals and is currently the most common parasite found in human stools. In some developing countries, its prevalence in study populations may exceed 50%. Morphologically, isolates of Blastocystis sp. found in different hosts are very similar. However, these same isolates show a very high genetic diversity between them and no less than 17 subtypes (or genotypes) have already been identified from molecular data. Genotyping studies have been carried out in many countries around the world and in particular in some Mediterranean countries such as France, spain, Italy, Turkey and Egypt. However, very little genotyping data is available in Algeria. To this end, we conducted the present study to identify and genotype Blastocystis in human and animal stool samples.Patients and methodsOne thousand eight hundred and sixty-nine (1,869) stool samples from kitchen staff as part of the periodic medical check-up, from subjects for the provision of a medical certificate required for the processing of a visa file and from patients with gastrointestinal disorders were examined. In addition to human faeces, animal samples, including 10 poultry, 2 cattle and 2 murine animals were examined. All stools were subjected to direct microscopic examination supplemented by concentration techniques and modified Ziehl Neelsen staining. Molecular characterization of 39 human and 14 animal isolates was performed by sequencing and the resulting sequences compared with those available from GenBank. Sequencing was only contributory for 30 human and 9 animal strains.ResultsOf all human samples examined 284 were positive (15.19%) with a prevalence of 7.38% for Blastocystis. Of the 30 strains that were molecularly characterized, ST3 was predominant (15/30, 50%) followed by ST1 (10/30, 33.33%) and in third place ST2 (4/30, 13.33%). ST4 was identified in only one patient (1/30, 3.33%). The correlation between clinical status and the subtype of Blastocystis identified showed that the number of ST3 was high in asymptomatic subjects (11/15, 73%) compared to symptomatic subjects (4/15, 26.66%), as well as for the ST1 subtype (7/10, 70% versus 3/10, 30%). Conversely, the number of ST2 was higher in subjects with gastrointestinal disorders (3/4, 75%). In addition to human strains, we genotyped 7 avian, 2 murine and 2 bovine strains. Characterization of the avian strains revealed 5 ST6 (71.42%) and 2 ST7 (28, 57%). The murine and bovine strains are identified as ST7 and ST6 respectively.

Dataset Information

Functional and Molecular Characterization of the Halomicrobium sp. IBSBa Inulosucrase.

Publications

Functional and Molecular Characterization of the <i>Halomicrobium</i> sp. IBSBa Inulosucrase.

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