Recombinant Production of Bifidobacterial Endoglycosidases for N-glycan Release
Küçük Resim Yok
Tarih
2021
Dergi Başlığı
Dergi ISSN
Cilt Başlığı
Yayıncı
Journal Of Visualized Experiments
Erişim Hakkı
info:eu-repo/semantics/closedAccess
Özet
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 nextgeneration 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.
Açıklama
Anahtar Kelimeler
Linked Glycans, Glycoproteins, Oligosaccharides, Deglycosylation, Acetylglucosaminidase, Glycosylation, Reveals
Kaynak
Jove-Journal of Visualized Experiments
WoS Q Değeri
Q3
Scopus Q Değeri
Q2
Cilt
Sayı
173
