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Analysis of free oligosaccharides (fOS) from wild-type saccharomyces cerevisiae (baker’s yeast) using two different extraction methods
Iqbal Jalaludin1, Amirul Husna Sudin2, Dharshini Elangovan3, Al-Bajalan, Hussein M4, Nur Maisarah Sarizan5, Noor Liana Mat Yajit6, Kamalrul Azlan Azizan7, Abdul Munir Abdul Murad8, Farah Diba Abu Bakar9, Alonzi, Dominic S10, Mukram Mohamad Mackeen11.
The glycomic profiles of free oligosaccharides (fOS) derived from misfolded N- and O-linked glycoproteins and lipidlinked 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 highperformance 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 HPLCELSD 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.
Affiliation:
- Universiti Kebangsaan Malaysia, Malaysia
- Universiti Kebangsaan Malaysia, Malaysia
- Universiti Kebangsaan Malaysia, Malaysia
- Universiti Kebangsaan Malaysia, Malaysia
- Universiti Kebangsaan Malaysia, Malaysia
- Universiti Kebangsaan Malaysia, Malaysia
- Universiti Kebangsaan Malaysia, Malaysia
- Universiti Kebangsaan Malaysia, Malaysia
- Universiti Kebangsaan Malaysia, Malaysia
- University of Oxford, United Kingdom
- Universiti Kebangsaan Malaysia, Malaysia
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6 |
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0.000 |
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0 |
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Q4 (Multidisciplinary Sciences) |
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Scopus 2020 |
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Q2 (Multidisciplinary) |
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