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Establishing a Metabolite Extraction Method to Study the Metabolome of Blastocystis Using NMR

Blastocystis is an opportunistic parasite commonly found in the intestines of humans and other animals. Despite its high prevalence, knowledge regarding Blastocystis biology within and outside the host is limited. Analysis of the metabolites produced by this anaerobe could provide insights that can...

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Autores principales: Newton, Jamie M., Betts, Emma L., Yiangou, Lyto, Ortega Roldan, Jose, Tsaousis, Anastasios D., Thompson, Gary S.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8199492/
https://www.ncbi.nlm.nih.gov/pubmed/34072445
http://dx.doi.org/10.3390/molecules26113285
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author Newton, Jamie M.
Betts, Emma L.
Yiangou, Lyto
Ortega Roldan, Jose
Tsaousis, Anastasios D.
Thompson, Gary S.
author_facet Newton, Jamie M.
Betts, Emma L.
Yiangou, Lyto
Ortega Roldan, Jose
Tsaousis, Anastasios D.
Thompson, Gary S.
author_sort Newton, Jamie M.
collection PubMed
description Blastocystis is an opportunistic parasite commonly found in the intestines of humans and other animals. Despite its high prevalence, knowledge regarding Blastocystis biology within and outside the host is limited. Analysis of the metabolites produced by this anaerobe could provide insights that can help map its metabolism and determine its role in both health and disease. Due to its controversial pathogenicity, these metabolites could define its deterministic role in microbiome’s “health” and/or subsequently resolve Blastocystis’ potential impact in gastrointestinal health. A common method for elucidating the presence of these metabolites is through (1)H nuclear magnetic resonance (NMR). However, there are currently no described benchmarked methods available to extract metabolites from Blastocystis for (1)H NMR analysis. Herein, several extraction solvents, lysis methods and incubation temperatures were compared for their usefulness as an extraction protocol for this protozoan. Following extraction, the samples were freeze-dried, re-solubilized and analysed with (1)H NMR. The results demonstrate that carrying out the procedure at room temperature using methanol as an extraction solvent and bead bashing as a lysis technique provides a consistent, reproducible and efficient method to extract metabolites from Blastocystis for NMR.
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spelling pubmed-81994922021-06-14 Establishing a Metabolite Extraction Method to Study the Metabolome of Blastocystis Using NMR Newton, Jamie M. Betts, Emma L. Yiangou, Lyto Ortega Roldan, Jose Tsaousis, Anastasios D. Thompson, Gary S. Molecules Article Blastocystis is an opportunistic parasite commonly found in the intestines of humans and other animals. Despite its high prevalence, knowledge regarding Blastocystis biology within and outside the host is limited. Analysis of the metabolites produced by this anaerobe could provide insights that can help map its metabolism and determine its role in both health and disease. Due to its controversial pathogenicity, these metabolites could define its deterministic role in microbiome’s “health” and/or subsequently resolve Blastocystis’ potential impact in gastrointestinal health. A common method for elucidating the presence of these metabolites is through (1)H nuclear magnetic resonance (NMR). However, there are currently no described benchmarked methods available to extract metabolites from Blastocystis for (1)H NMR analysis. Herein, several extraction solvents, lysis methods and incubation temperatures were compared for their usefulness as an extraction protocol for this protozoan. Following extraction, the samples were freeze-dried, re-solubilized and analysed with (1)H NMR. The results demonstrate that carrying out the procedure at room temperature using methanol as an extraction solvent and bead bashing as a lysis technique provides a consistent, reproducible and efficient method to extract metabolites from Blastocystis for NMR. MDPI 2021-05-29 /pmc/articles/PMC8199492/ /pubmed/34072445 http://dx.doi.org/10.3390/molecules26113285 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
Newton, Jamie M.
Betts, Emma L.
Yiangou, Lyto
Ortega Roldan, Jose
Tsaousis, Anastasios D.
Thompson, Gary S.
Establishing a Metabolite Extraction Method to Study the Metabolome of Blastocystis Using NMR
title Establishing a Metabolite Extraction Method to Study the Metabolome of Blastocystis Using NMR
title_full Establishing a Metabolite Extraction Method to Study the Metabolome of Blastocystis Using NMR
title_fullStr Establishing a Metabolite Extraction Method to Study the Metabolome of Blastocystis Using NMR
title_full_unstemmed Establishing a Metabolite Extraction Method to Study the Metabolome of Blastocystis Using NMR
title_short Establishing a Metabolite Extraction Method to Study the Metabolome of Blastocystis Using NMR
title_sort establishing a metabolite extraction method to study the metabolome of blastocystis using nmr
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8199492/
https://www.ncbi.nlm.nih.gov/pubmed/34072445
http://dx.doi.org/10.3390/molecules26113285
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