Recombinant Production of Bifidobacterial Endoglycosidases for N-glycan Release.

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ID: 270046
2021
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Abstract
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.
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Authors Sucu, Berfin;Bayraktar, Ayşe;Duman, Hatice;Arslan, Ayşenur;Kaplan, Merve;Karyelioğlu, Melda;Ntelitze, Eda;Taştekin, Taner;Yetkin, Seray;Ertürk, Melih;Frese, Steven A;Henrick, Bethany M;Kayili, Hacı Mehmet;Salih, Bekir;Karav, Sercan;
Journal Journal of visualized experiments : JoVE
Year 2021
DOI
10.3791/62804
URL
Keywords
prediction model casualties pandemic covid-19 linear dynamics non-linear dynamics

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