Establishing a Metabolite Extraction Method to Study the Metabolome of <i>Blastocystis</i> Using NMR
<i>Blastocystis</i> is an opportunistic parasite commonly found in the intestines of humans and other animals. Despite its high prevalence, knowledge regarding <i>Blastocystis</i> biology within and outside the host is limited. Analysis of the metabolites produced by this ana...
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| Format: | Article |
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MDPI AG
2021-05-01
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| Series: | Molecules |
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| Online Access: | https://www.mdpi.com/1420-3049/26/11/3285 |
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| author | Jamie M. Newton Emma L. Betts Lyto Yiangou Jose Ortega Roldan Anastasios D. Tsaousis Gary S. Thompson |
| author_facet | Jamie M. Newton Emma L. Betts Lyto Yiangou Jose Ortega Roldan Anastasios D. Tsaousis Gary S. Thompson |
| author_sort | Jamie M. Newton |
| collection | DOAJ |
| description | <i>Blastocystis</i> is an opportunistic parasite commonly found in the intestines of humans and other animals. Despite its high prevalence, knowledge regarding <i>Blastocystis</i> 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 <i>Blastocystis</i>’ potential impact in gastrointestinal health. A common method for elucidating the presence of these metabolites is through <sup>1</sup>H nuclear magnetic resonance (NMR). However, there are currently no described benchmarked methods available to extract metabolites from <i>Blastocystis</i> for <sup>1</sup>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 <sup>1</sup>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 <i>Blastocystis</i> for NMR. |
| first_indexed | 2024-03-10T10:53:17Z |
| format | Article |
| id | doaj.art-0e19f48d21a246778d336ab30cd5e131 |
| institution | Directory Open Access Journal |
| issn | 1420-3049 |
| language | English |
| last_indexed | 2024-03-10T10:53:17Z |
| publishDate | 2021-05-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Molecules |
| spelling | doaj.art-0e19f48d21a246778d336ab30cd5e1312023-11-21T22:00:18ZengMDPI AGMolecules1420-30492021-05-012611328510.3390/molecules26113285Establishing a Metabolite Extraction Method to Study the Metabolome of <i>Blastocystis</i> Using NMRJamie M. Newton0Emma L. Betts1Lyto Yiangou2Jose Ortega Roldan3Anastasios D. Tsaousis4Gary S. Thompson5Laboratory of Molecular & Evolutionary Parasitology, RAPID Group, School of Biosciences, University of Kent, Canterbury CT2 7NJ, UKLaboratory of Molecular & Evolutionary Parasitology, RAPID Group, School of Biosciences, University of Kent, Canterbury CT2 7NJ, UKLaboratory of Molecular & Evolutionary Parasitology, RAPID Group, School of Biosciences, University of Kent, Canterbury CT2 7NJ, UKWellcome Trust Biomolecular NMR Facility, School of Biosciences, University of Kent, Canterbury CT2 7NJ, UKLaboratory of Molecular & Evolutionary Parasitology, RAPID Group, School of Biosciences, University of Kent, Canterbury CT2 7NJ, UKWellcome Trust Biomolecular NMR Facility, School of Biosciences, University of Kent, Canterbury CT2 7NJ, UK<i>Blastocystis</i> is an opportunistic parasite commonly found in the intestines of humans and other animals. Despite its high prevalence, knowledge regarding <i>Blastocystis</i> 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 <i>Blastocystis</i>’ potential impact in gastrointestinal health. A common method for elucidating the presence of these metabolites is through <sup>1</sup>H nuclear magnetic resonance (NMR). However, there are currently no described benchmarked methods available to extract metabolites from <i>Blastocystis</i> for <sup>1</sup>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 <sup>1</sup>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 <i>Blastocystis</i> for NMR.https://www.mdpi.com/1420-3049/26/11/3285<i>Blastocystis</i><sup>1</sup>H NMRmetabolite extraction, metabolomics |
| spellingShingle | Jamie M. Newton Emma L. Betts Lyto Yiangou Jose Ortega Roldan Anastasios D. Tsaousis Gary S. Thompson Establishing a Metabolite Extraction Method to Study the Metabolome of <i>Blastocystis</i> Using NMR Molecules <i>Blastocystis</i> <sup>1</sup>H NMR metabolite extraction, metabolomics |
| title | Establishing a Metabolite Extraction Method to Study the Metabolome of <i>Blastocystis</i> Using NMR |
| title_full | Establishing a Metabolite Extraction Method to Study the Metabolome of <i>Blastocystis</i> Using NMR |
| title_fullStr | Establishing a Metabolite Extraction Method to Study the Metabolome of <i>Blastocystis</i> Using NMR |
| title_full_unstemmed | Establishing a Metabolite Extraction Method to Study the Metabolome of <i>Blastocystis</i> Using NMR |
| title_short | Establishing a Metabolite Extraction Method to Study the Metabolome of <i>Blastocystis</i> Using NMR |
| title_sort | establishing a metabolite extraction method to study the metabolome of i blastocystis i using nmr |
| topic | <i>Blastocystis</i> <sup>1</sup>H NMR metabolite extraction, metabolomics |
| url | https://www.mdpi.com/1420-3049/26/11/3285 |
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