Recombinant Production and Characterization of a Novel N-glycosidases (EndoBI-2) from Bifidobacterium longum subsp. infantis 157F


Duman H., Karav S.

5th International Eurasian Conference on Biological and Chemical Sciences (EurasianBioChem), 2022, 23 November 2022, pp.293

  • Publication Type: Conference Paper / Summary Text
  • Page Numbers: pp.293
  • Çanakkale Onsekiz Mart University Affiliated: Yes

Abstract

The gastrointestinal (GI) tract harbors several microorganisms, which contribute considerably to the maintenance of health and the progression of diseases. The survival of microorganisms that live in the GI tract is mainly dependent on their ability to utilize complex carbohydrates (glycans and polysaccharides) that are not digestible by the host organism. Human milk is the primary source of nutrition for a newborn before they can digest any other food. Glycosylated milk components such as lactoferrin, immunoglobulins, etc., are indigestible to the infant, and only certain gut microbes have the enzymes to release glycans from glycoproteins and utilize these compounds as the only carbon source. Glycosidases are the type of enzymes that are used for N-glycan release from glycoproteins. Commercially available glycosidases are inefficient at cleaving these glycans from proteins in their native state. To overcome this deficiency, efficient deglycosylation approaches are required to better understand the biological roles of N-glycans and their large-scale production. A novel enzyme, endo-ß-N-acetylglucosaminidase 2 (EndoBI-2) was isolated from Bifidobacterium longum subsp. infantis is a unique and effective enzyme for this process, but this enzyme isn’t commercially available. Within this study, the novel enzyme was produced and characterized recombinantly to understand the unique features of N-glycans. Comparative modeling approaches were used to investigate the structural properties and substrate selectivity of EndoBI-2. Furthermore, the optimum reaction/kinetic parameters of EndoBI-2 were investigated using glycoproteins and released bioactive glycans were characterized. The kinetic and structural characterization parameters of EndoBI-2 suggest that this novel enzyme can facilitate the largescale release of bioactive glycans from glycoprotein sources. Successful application of this novel enzyme and released bioactive glycans in the food industry will enable to development of functional foods for health and wellbeing.