TY - JOUR
T1 - Characterization of novel endo-β-N-acetylglucosaminidase from Bacteroides nordii that hydrolyzes multi-branched complex type N-glycans
AU - Bienes, Kristina Mae
AU - Tautau, Feunai Agape Papalii
AU - Mitani, Ai
AU - Kinoshita, Takashi
AU - Nakakita, Shin ichi
AU - Higuchi, Yujiro
AU - Takegawa, Kaoru
N1 - Funding Information:
The authors declare no conflict of interest. This work was partially supported by the Japan Society for the Promotion of Science (JSPS) KAKENHI grant number JP21K19090, and the Institute for Fermentation Osaka (IFO), Japan (KT).
Publisher Copyright:
© 2022 The Society for Biotechnology, Japan
PY - 2022/7
Y1 - 2022/7
N2 - 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.
AB - 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.
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U2 - 10.1016/j.jbiosc.2022.03.011
DO - 10.1016/j.jbiosc.2022.03.011
M3 - Article
C2 - 35484013
AN - SCOPUS:85129903718
SN - 1389-1723
VL - 134
SP - 7
EP - 13
JO - Journal of Bioscience and Bioengineering
JF - Journal of Bioscience and Bioengineering
IS - 1
ER -