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Syntheses of mucin-type O-glycopeptides and oligosaccharides using transglycosylation and reverse-hydrolysis activities of Bifidobacterium endo-α-N-acetylgalactosaminidase

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Abstract

Endo-α-N-acetylgalactosaminidase catalyzes the release of Galβ1-3GalNAc from the core 1-type O-glycan (Galβ1-3GalNAcα1-Ser/Thr) of mucin glycoproteins and synthetic p-nitrophenyl (pNP) α-linked substrates. Here, we report the enzymatic syntheses of core 1 disaccharide-containing glycopeptides using the transglycosylation activity of endo-α-N-acetylgalactosaminidase (EngBF) from Bifidobacterium longum. The enzyme directly transferred Galβ1-3GalNAc to serine or threonine residues of bioactive peptides such as PAMP-12, bradykinin, peptide-T and MUC1a when Galβ1-3GalNAcα1-pNP was used as a donor substrate. The enzyme was also found to catalyze the reverse-hydrolysis reaction. EngBF synthesized the core 1 disaccharide-containing oligosaccharides when the enzyme was incubated with either glucose or lactose and Galβ1-3GalNAc prepared from porcine gastric mucin using bifidobacterial cells expressing endo-α-N-acetylgalactosaminidase. Synthesized oligosaccharides are promising prebiotics for bifidobacteria.

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Acknowledgments

We thank Dr. H. Ishida and Dr. M. Kiso (Gifu University), Dr. M. Yamaguchi (Wakayama University) and Dr. T. Yamanoi (the Noguchi Institute) for MS analyses. This work was supported in part by grant-in-aids from the Promotion of Basic Research Activities for Innovative Biosciences (PROBRAIN) and from Ajinomoto Co., Inc.

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Authors and Affiliations

  1. Graduate School of Biostudies, Kyoto University, Kyoto, 606-8502, Japan

    Hisashi Ashida, Hayato Ozawa & Kenji Yamamoto

  2. Faculty of Agriculture, Kagoshima University, Kagoshima, 890-0065, Japan

    Kiyotaka Fujita

  3. Ajinomoto Co., Inc., Kawasaki, Kanagawa, 210-8681, Japan

    Shun’ichi Suzuki

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  1. Hisashi Ashida
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  2. Hayato Ozawa
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  3. Kiyotaka Fujita
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  4. Shun’ichi Suzuki
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  5. Kenji Yamamoto
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Correspondence to Hisashi Ashida.

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Ashida, H., Ozawa, H., Fujita, K. et al. Syntheses of mucin-type O-glycopeptides and oligosaccharides using transglycosylation and reverse-hydrolysis activities of Bifidobacterium endo-α-N-acetylgalactosaminidase. Glycoconj J 27, 125–132 (2010). https://doi.org/10.1007/s10719-009-9247-8

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  • DOI: https://doi.org/10.1007/s10719-009-9247-8

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