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HPLC Profiling of N-Glycans from Ganoderma boninense released by peptide N-Glycosidase (PNGaseF)

Sarizan, Nur Maisarah; Allman, Sarah; Needs, Sarah; Han, Goh Hoe; Murad, Abdul Munir Abdul; Bakar, Farah Diba Abu; Seman, Idris Abu and Mackeen, Mukram Mohamed (2017). HPLC Profiling of N-Glycans from Ganoderma boninense released by peptide N-Glycosidase (PNGaseF). In: International Conference on Natural Products 2017, 15-16 Mar 2017, Swiss-Garden Beach Resort Damai Laut, Lumut, Malaysia.

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Abstract

Asparagine (N)-linked glycosylation is the most common modification found on proteins. However, the structures of carbohydrates/glycans present in the glycoproteins are difficult to analyse due to the presence of multiple stereo- and regio-isomers. The analysis of glycan structures is aided by the release of glycans from glycoproteins using the enzyme of peptide N-glycosidase F (PNGase F). A previously prepared PNGase F plasmid was successfully expressed and purified, and the activity of the enzyme showed that PNGase F was able to deglycosylate thea standard substrate, ribonuclease B. The cell pellets of Ganoderma boninense, a major pathogen of oil palm, was also digested with the purified PNGase F. The released N-glycans were labelled with the fluorophore 2-aminobenzamide (2-AB) and analysed using high performance liquid chromatography with fluorescence detection (HPLC-FD). Peaks observed in the G. boninense samples were found to range from glucose unit 1 (GU1) until up to more than GU16 (limit of detection) when compared to a dextran oligomer ladder. The highest peaks in G.boninense were from GU1 to GU4 indicating of the high abundance of small glycans (mono- to tetrasaccharides). Additionally, there are many small peaks observed at GU14 and above suggesting the possible occurrence of hyperglycosylation.

Item Type: Conference or Workshop Item
Academic Unit/School: Faculty of Science, Technology, Engineering and Mathematics (STEM) > Life, Health and Chemical Sciences
Faculty of Science, Technology, Engineering and Mathematics (STEM)
Interdisciplinary Research Centre: Biomedical Research Network (BRN)
Related URLs:
Item ID: 49043
Depositing User: Sarah Allman
Date Deposited: 29 Mar 2017 08:50
Last Modified: 29 Mar 2017 08:50
URI: http://oro.open.ac.uk/id/eprint/49043
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