Analysis of free oligosaccharides (fOS) from wild-type Saccharomyces cerevisiae (Baker’s yeast) using two different extraction methods
The glycomic profiles of free oligosaccharides (fOS) derived from misfolded N- and O-linked glycoproteins and lipid-linked oligosaccharides are important molecular signatures in various biological processes and serve as a readout of functional properties such as glycosidase inhibition. Several glyca...
| Main Authors: | , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Penerbit Universiti Kebangsaan Malaysia
2020
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| Online Access: | http://journalarticle.ukm.my/14734/1/ARTIKEL%2010.pdf |
| _version_ | 1796932129544732672 |
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| author | Iqbal Jalaludin, Amirul Husna Sudin, Dharshini Elangovan, Hussein M. Al-Bajalan, Nur Maisarah Sarizan, Noor Liana Mat Yajit, Kamalrul Azlan Azizan, Abdul Munir Abdul Murad, Farah Diba Abu Bakar, Alonzi, Dominic S. Mukram Mohamad Mackeen, |
| author_facet | Iqbal Jalaludin, Amirul Husna Sudin, Dharshini Elangovan, Hussein M. Al-Bajalan, Nur Maisarah Sarizan, Noor Liana Mat Yajit, Kamalrul Azlan Azizan, Abdul Munir Abdul Murad, Farah Diba Abu Bakar, Alonzi, Dominic S. Mukram Mohamad Mackeen, |
| author_sort | Iqbal Jalaludin, |
| collection | UKM |
| description | The glycomic profiles of free oligosaccharides (fOS) derived from misfolded N- and O-linked glycoproteins and lipid-linked oligosaccharides are important molecular signatures in various biological processes and serve as a readout of functional properties such as glycosidase inhibition. Several glycan extraction methods are available based on different sorbent chemistries that may influence the analytical profiles obtained. However, there is limited availability of studies comparing the effects of sorbent chemistries on glycan profiles. Therefore, in our study, the fOS profiles from wild-type Saccharomyces cerevisiae (Baker’s yeast) extracted using two common methods namely mixed-bed ion-exchange (MBIE) [AG50W-X12 (H+) and AG2-X8 (Cl-)] and reversed-phase (C18) sorbents were compared using total carbohydrate (phenol sulfuric acid) and total protein (bicinchoninic acid, BCA) assays, thin-layer chromatography (TLC) and high-performance liquid chromatography-evaporative light scattering detector (HPLC-ELSD) analyses. MBIE extraction contained higher oligosaccharide and protein (0.26 mg/mL and 1.8 mg/mL) content than C18 extraction (0.11 mg/mL and 0.2 mg/mL). TLC analysis (butanol: ethanol: water = 6:3:1 and 5:4:1) showed the presence of fOS in both the MBIE and C18 extracts based on the detection of orcinol active (UV-inactive) spots. Similar peaks were present in the HPLC-ELSD chromatograms for both extractions methods with MBIE showing higher abundance. Glycan unit (GU) analysis of the dextran standard using HPLC-ELSD showed that the largest possible oligosaccharide structures detected were only di/trisaccharides. Based on all these results, MBIE extraction is a more suitable carbohydrate extraction technique compared to C18 extraction for subsequent profiling and functional studies of fOS. |
| first_indexed | 2024-03-06T04:28:14Z |
| format | Article |
| id | ukm.eprints-14734 |
| institution | Universiti Kebangsaan Malaysia |
| language | English |
| last_indexed | 2024-03-06T04:28:14Z |
| publishDate | 2020 |
| publisher | Penerbit Universiti Kebangsaan Malaysia |
| record_format | dspace |
| spelling | ukm.eprints-147342020-06-17T07:50:32Z http://journalarticle.ukm.my/14734/ Analysis of free oligosaccharides (fOS) from wild-type Saccharomyces cerevisiae (Baker’s yeast) using two different extraction methods Iqbal Jalaludin, Amirul Husna Sudin, Dharshini Elangovan, Hussein M. Al-Bajalan, Nur Maisarah Sarizan, Noor Liana Mat Yajit, Kamalrul Azlan Azizan, Abdul Munir Abdul Murad, Farah Diba Abu Bakar, Alonzi, Dominic S. Mukram Mohamad Mackeen, The glycomic profiles of free oligosaccharides (fOS) derived from misfolded N- and O-linked glycoproteins and lipid-linked oligosaccharides are important molecular signatures in various biological processes and serve as a readout of functional properties such as glycosidase inhibition. Several glycan extraction methods are available based on different sorbent chemistries that may influence the analytical profiles obtained. However, there is limited availability of studies comparing the effects of sorbent chemistries on glycan profiles. Therefore, in our study, the fOS profiles from wild-type Saccharomyces cerevisiae (Baker’s yeast) extracted using two common methods namely mixed-bed ion-exchange (MBIE) [AG50W-X12 (H+) and AG2-X8 (Cl-)] and reversed-phase (C18) sorbents were compared using total carbohydrate (phenol sulfuric acid) and total protein (bicinchoninic acid, BCA) assays, thin-layer chromatography (TLC) and high-performance liquid chromatography-evaporative light scattering detector (HPLC-ELSD) analyses. MBIE extraction contained higher oligosaccharide and protein (0.26 mg/mL and 1.8 mg/mL) content than C18 extraction (0.11 mg/mL and 0.2 mg/mL). TLC analysis (butanol: ethanol: water = 6:3:1 and 5:4:1) showed the presence of fOS in both the MBIE and C18 extracts based on the detection of orcinol active (UV-inactive) spots. Similar peaks were present in the HPLC-ELSD chromatograms for both extractions methods with MBIE showing higher abundance. Glycan unit (GU) analysis of the dextran standard using HPLC-ELSD showed that the largest possible oligosaccharide structures detected were only di/trisaccharides. Based on all these results, MBIE extraction is a more suitable carbohydrate extraction technique compared to C18 extraction for subsequent profiling and functional studies of fOS. Penerbit Universiti Kebangsaan Malaysia 2020-01 Article PeerReviewed application/pdf en http://journalarticle.ukm.my/14734/1/ARTIKEL%2010.pdf Iqbal Jalaludin, and Amirul Husna Sudin, and Dharshini Elangovan, and Hussein M. Al-Bajalan, and Nur Maisarah Sarizan, and Noor Liana Mat Yajit, and Kamalrul Azlan Azizan, and Abdul Munir Abdul Murad, and Farah Diba Abu Bakar, and Alonzi, Dominic S. and Mukram Mohamad Mackeen, (2020) Analysis of free oligosaccharides (fOS) from wild-type Saccharomyces cerevisiae (Baker’s yeast) using two different extraction methods. Sains Malaysiana, 49 (1). pp. 85-92. ISSN 0126-6039 http://www.ukm.my/jsm/malay_journals/jilid49bil1_2020/KandunganJilid49Bil1_2020.html |
| spellingShingle | Iqbal Jalaludin, Amirul Husna Sudin, Dharshini Elangovan, Hussein M. Al-Bajalan, Nur Maisarah Sarizan, Noor Liana Mat Yajit, Kamalrul Azlan Azizan, Abdul Munir Abdul Murad, Farah Diba Abu Bakar, Alonzi, Dominic S. Mukram Mohamad Mackeen, Analysis of free oligosaccharides (fOS) from wild-type Saccharomyces cerevisiae (Baker’s yeast) using two different extraction methods |
| title | Analysis of free oligosaccharides (fOS) from wild-type Saccharomyces cerevisiae (Baker’s yeast) using two different extraction methods |
| title_full | Analysis of free oligosaccharides (fOS) from wild-type Saccharomyces cerevisiae (Baker’s yeast) using two different extraction methods |
| title_fullStr | Analysis of free oligosaccharides (fOS) from wild-type Saccharomyces cerevisiae (Baker’s yeast) using two different extraction methods |
| title_full_unstemmed | Analysis of free oligosaccharides (fOS) from wild-type Saccharomyces cerevisiae (Baker’s yeast) using two different extraction methods |
| title_short | Analysis of free oligosaccharides (fOS) from wild-type Saccharomyces cerevisiae (Baker’s yeast) using two different extraction methods |
| title_sort | analysis of free oligosaccharides fos from wild type saccharomyces cerevisiae baker s yeast using two different extraction methods |
| url | http://journalarticle.ukm.my/14734/1/ARTIKEL%2010.pdf |
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