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Tüberküloz, bulaşıcı hastalıklara bağlı ölümlerin başlıca nedenlerinden biridir. Tüberküloz tedavisinin uzun ve karmaşık olması, ilaç direncini yaygınlaşmaktadır. Tüberküloz tedavisinde umut olacak bazı benzimidazol türevi bileşiklerin antitüberküloz aktivite sergilediği bildirilmiştir. Çalışmada, referans M. tuberculosis H37Rv suşuna karşı sentezlenen benzimidazol çekirdeği içeren 3 farklı bileşiğin (A1: 1-(2-metilbenzonitril)-3-(naftalen-1-il-metil)-1H-benzo[d]imidazol-3-yum klorür, A2: 1-(2-hidroksietil)-3-(2-(piperidinyum-1-il)etil klorür)-1H-benzo[d]imidazol-3-yum bromür, A3: 1-(3-metilbenzil)-3-(2-(piperidin-1-il)etil)-1H-benzo[d]imidazol-3-yum klorür) antitüberküloz aktivitesinin tespiti amaçlandı. Benzimidazol çekirdeği içeren bileşiklerin (A1, A2 ve A3), M. tuberculosis H37Rv suşuna karşı antitüberküloz aktiviteleri in vitro şartlarda BACTEC MGIT 960 sistemi kullanılarak araştırıldı. Çalışmada A1, A2 ve A3 bileşiklerinin M. tuberculosis H37Rv suşuna karşı herhangi bir antitüberküloz aktivite göstermediği tespit edildi. Sonuç olarak, farklı benzimidazol bileşikler sentezlenerek antitüberküloz aktivite çalışmalarının devam edilmesi gerektiği düşünülmektedir.

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.

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.

Complex, indigestible free oligosaccharides as well as conjugated glycans are found in milk that shape the gut microbiome of infants. The activity of an endo-β-N-acetylglucosaminidase from B. longum subsp. infantis (B. infantis) is known to release N-glycans from native milk glycoproteins under physiological conditions. We investigated whether this enzyme is active in vivo in breastfed infants fed B. infantis EVC001. Using mass spectrometry, we found 19 N-glycans related to human milk glycoproteins increased in abundance, similar to previous work using bovine milk glycoproteins, and these 19 N-glycans matched unique specificities of this enzyme. Twenty N-glycans were unique to infants fed B. infantis EVC001. Bifidobacteriaceae were correlated with these glycans, confirming the relationship between B. infantis and released N-glycans. This suggests that this enzyme is active in vivo and releases N-glycans from milk glycoproteins, and may play a role in B. infantis EVC001 colonization of the gut microbiome.