{Reference Type}: Journal Article
{Title}: Characterization of novel endo-β-N-acetylglucosaminidase from Bacteroides nordii that hydrolyzes multi-branched complex type N-glycans.
{Author}: Bienes KM;Tautau FAP;Mitani A;Kinoshita T;Nakakita SI;Higuchi Y;Takegawa K;Bienes KM;Tautau FAP;Mitani A;Kinoshita T;Nakakita SI;Higuchi Y;Takegawa K;
{Journal}: J Biosci Bioeng
{Volume}: 134
{Issue}: 1
{Year}: Jul 2022
{Factor}: 3.185
{DOI}: 10.1016/j.jbiosc.2022.03.011
{Abstract}: Endo-β-N-acetylglucosaminidases (ENGases) are enzymes that hydrolyze the N-linked oligosaccharides. Many ENGases have already been identified and characterized. However, there are still a few enzymes that have hydrolytic activity toward multibranched complex-type N-glycans on glycoproteins. In this study, one novel ENGase from Bacteroides nordii (Endo-BN) species was identified and characterized. The recombinant protein was prepared and expressed in Escherichia coli cells. This Endo-BN exhibited optimum hydrolytic activity at pH 4.0. High performance liquid chromatography (HPLC) analysis showed that Endo-BN preferred core-fucosylated complex-type N-glycans, with galactose or α2,6-linked sialic acid residues at their non-reducing ends. The hydrolytic activities of Endo-BN were also tested on different glycoproteins from high-mannose type to complex-type oligosaccharides. The reaction with human transferrin, fetuin, and α1-acid glycoprotein subsequently showed that Endo-BN is capable of releasing multi-branched complex-type N-glycans from these glycoproteins.