IV-INTERNATIONAL CONFERENCE OF FOOD, AGRICULTURE, AND VETERINARY SCIENCES, Van, Turkey, 27 - 28 May 2022, pp.426-427
ABSTRACT Edible mushrooms have been consumed since ancient times and are classified as a special food. Except for their special aroma, they have high nutritional quality and contain many different bioactive compounds. It has been found that they contain a high amount of proteins and essential amino acids which give them many health-beneficial features such as anticancer, antibiotic, and antioxidant properties. The protein content of each mushroom species depends on the species and affects its health and beneficial properties. Therefore, it is important to investigate each protein fragment of different mushroom species separately. Fungal proteins of each species are isolated and visualized by SDS-PAGE gel electrophoresis and their protein content is analyzed using Qubit 3.0 fluorometric analysis. Protein glycosylation is a post-translational modification that affects protein activity, stability, and protein-protein interactions. Glycans affect the stabilization, conformation, and folding properties of proteins. In order to examine the effects of glycans and to understand their functions, they must be separated from the proteins to which they are attached.
Deglycosylation is a process of separating glycans from glycoproteins. Peptide- N- glucosidase F (PNGase F) is a mainly used enzyme that catalyzes the deglycosylation of N- glycans from glycoproteins. Recently, it has been reported that free N- glycans can be used as prebiotic sources. This study is aimed to deglycolyze glycoproteins of three different edible mushroom species(Marasmius Castaneophilus, Agaricus Bisporus, Pleurotus Ostreatus) by using the PNGase F enzyme. The enzyme cleaves the fungal glycoproteins and free N- glycans are obtained. Characterization of these N- glycans will be done by using MALDI-TOF Mass spectrometry. Obtained N- glycans are potential prebiotic candidates and their prebiotic activity will be tested by using an in-vitro digestion system. Keywords: Edible mushroom, N- glycan, MALDI TOF, prebiotic, PNGase F.
*This study was derived from the Graduate Thesis of Tuba ÇAĞIRTEKİN entitled as “Protein Isolation and N- Glycan Characterization of Different Mushroom Species by Using PNGase F enzyme” supplied for partial fulfillment of the Master’s Degree at Molecular Biology and Genetics Department at School of Graduate Studies at Çanakkale Onsekiz Mart University. The study was financially supported by Çanakkale Onsekiz Mart University Scientific Research Projects Department with the project number of FYL-2022-3873.