| Autor: |
Sucu B; Department of Molecular Biology and Genetics, Çanakkale Onsekiz Mart University., Bayraktar A; Department of Molecular Biology and Genetics, Çanakkale Onsekiz Mart University; Uluova Dairy Farm., Duman H; Department of Molecular Biology and Genetics, Çanakkale Onsekiz Mart University., Arslan A; Department of Molecular Biology and Genetics, Çanakkale Onsekiz Mart University., Kaplan M; Department of Molecular Biology and Genetics, Çanakkale Onsekiz Mart University., Karyelioğlu M; Department of Molecular Biology and Genetics, Çanakkale Onsekiz Mart University., Ntelitze E; Department of Molecular Biology and Genetics, Çanakkale Onsekiz Mart University., Taştekin T; Department of Molecular Biology and Genetics, Çanakkale Onsekiz Mart University., Yetkin S; Department of Molecular Biology and Genetics, Çanakkale Onsekiz Mart University., Ertürk M; Uluova Dairy Farm., Frese SA; Department of Nutrition, University of Nevada; Department of Food Science and Technology, University of Nebraska., Henrick BM; Department of Food Science and Technology, University of Nebraska; Evolve BioSystems Inc., Kayili HM; Department of Biomedical Engineering, Karabuk University., Salih B; Department of Chemistry, Hacettepe University., Karav S; Department of Molecular Biology and Genetics, Çanakkale Onsekiz Mart University; sercankarav@comu.edu.tr. |
| Abstrakt: |
Protein glycosylation is a diverse and common post-translational modification that has been associated with many important roles such as protein function, including protein folding, stability, enzymatic protection, and biological recognition. N-glycans attached to glycoproteins (such as lactoferrin, lactadherin, and immunoglobulins) cannot be digested by the host and reach the large intestine, where they are consumed by certain beneficial microbes. Therefore, they are considered next-generation prebiotic compounds that can selectively stimulate the gut microbiome's beneficial microorganisms. However, the isolation of these new classes of prebiotics requires novel enzymes. Here, we describe the recombinant production of novel glycosidases from different Bifidobacteria strains (isolated from infants, rabbits, chicken, and bumblebee) for improved N-glycan isolation from glycoproteins. The method presented in this study includes the following steps: molecular cloning of Bifidobacterial genes by an in vivo recombinational cloning strategy, control of transformation success, protein induction, and protein purification. |
