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Glycoconjugate Journal

, Volume 35, Issue 1, pp 31–40 | Cite as

Enzymatic synthesis of fucose-containing galacto-oligosaccharides using β-galactosidase and identification of novel disaccharide structures

  • Anne Usvalampi
  • Hannu Maaheimo
  • Olli Tossavainen
  • Alexander D. Frey
Original Article

Abstract

Fucosylated oligosaccharides have an important role in maintaining a healthy immune system and homeostatic gut microflora. This study employed a commercial β-galactosidase in the production of fucose-containing galacto-oligosaccharides (fGOS) from lactose and fucose. The production was optimized using experiment design and optimal conditions for a batch production in 3-liter scale. The reaction product was analyzed and the produced galactose-fucose disaccharides were purified. The structures of these disaccharides were determined using NMR and it was verified that one major product with the structure Galβ1–3Fuc and two minor products with the structures Galβ1–4Fuc and Galβ1–2Fuc were formed. Additionally, the product composition was defined in more detail using several different analytical methods. It was concluded that the final product contained 42% total monosaccharides, 40% disaccharides and 18% of larger oligosaccharides. 290 μmol of fGOS was produced per gram of reaction mixture and 37% of the added fucose was bound to fGOS. The fraction of fGOS from total oligosaccharides was determined as 44%. This fGOS product could be used as a new putative route to deliver fucose to the intestine.

Keywords

β-galactosidase Transglycosylation Fucose Hetero-oligosaccharide Galacto-oligosaccharide 

Notes

Compliance with ethical standards

Conflicts of interest

The authors declare that they have no conflicts of interest.

Ethical approval

This article does not contain any studies with human participants or animals performed by any of the authors.

Supplementary material

10719_2017_9794_MOESM1_ESM.pdf (387 kb)
ESM 1 (PDF 386 kb)

References

  1. 1.
    Bode, L.: Recent advances on structure, metabolism, and function of human milk oligosaccharides. J. Nutr. 136, 2127–2130 (2006)CrossRefPubMedGoogle Scholar
  2. 2.
    Oliveira, D.L., Wilbey, R.A., Grandison, A.S., Roseiro, L.B.: Milk oligosaccharides: a review. Int. J. Dairy Technol. 68, 305–321 (2015)CrossRefGoogle Scholar
  3. 3.
    Ninonuevo, M.R., Park, Y., Yin, H., Zhang, J., Ward, R.E., Clowers, B.H., German, J.B., Freeman, S.L., Killeen, K., Grimm, R., Lebrilla, C.B.: A strategy for annotating the human milk glycome. J. Agric. Food Chem. 54, 7471–7480 (2006)CrossRefPubMedGoogle Scholar
  4. 4.
    Cooling, L.: Blood groups in infection and host susceptibility. Clin. Microbiol. Rev. 28, 801–870 (2015)CrossRefPubMedPubMedCentralGoogle Scholar
  5. 5.
    Goto, Y., Uematsu, S., Kiyono, H.: Epithelial glycosylation in gut homeostasis and inflammation. Nature Immunol. 17, 1244–1251 (2016)CrossRefGoogle Scholar
  6. 6.
    Pickard, J.M., Chervonsky, A.V.: Intestinal fucose as a mediator of host-microbe symbiosis. J. Immunol. 194, 5588–5593 (2015)CrossRefPubMedPubMedCentralGoogle Scholar
  7. 7.
    Chow, W.L., Lee, Y.K.: Free fucose is a danger signal to human intestinal epithelial cells. Br. J. Nutr. 99, 449–454 (2008)CrossRefPubMedGoogle Scholar
  8. 8.
    Bode, L.: Human milk oligosaccharides: every baby needs a sugar mama. Glycobiology. 22, 1147–1162 (2012)CrossRefPubMedPubMedCentralGoogle Scholar
  9. 9.
    Barile, D., Rastall, R.A.: Human milk and related oligosaccharides as prebiotics. Curr. Opin. Biotechnol. 24, 214–219 (2013)CrossRefPubMedGoogle Scholar
  10. 10.
    Hickey, R.M.: The role of oligosaccharides from human milk and other sources in prevention of pathogen adhesion. Int. Dairy J. 22, 141–146 (2012)CrossRefGoogle Scholar
  11. 11.
    Lu, L.L., Xiao, M., Li, Z.Y., Li, Y.M., Wang, F.S.: A novel transglycosylating β-galactosidase from Enterobacter Cloacae B5. Process Biochem. 44, 232–236 (2009)CrossRefGoogle Scholar
  12. 12.
    Gosling, A., Stevens, G.W., Barber, A.R., Kentish, S.E., Gras, S.L.: Recent advances refining galactooligosaccharide production from lactose. Food Chem. 121, 307–318 (2010)CrossRefGoogle Scholar
  13. 13.
    Gänzle, M.G.: Enzymatic synthesis of galacto-oligosaccharides and other lactose derivatives (hetero-oligosaccharides) from lactose. Int. Dairy J. 22, 116–122 (2012)CrossRefGoogle Scholar
  14. 14.
    Li, W., Sun, Y., Ye, H., Zeng, X.: Synthesis of oligosaccharides with lactose and N-acetylglucosamine as substrates by using β-d-galactosidase from Bacillus Circulans. Eur. Food Res. Technol. 231, 55–63 (2010)CrossRefGoogle Scholar
  15. 15.
    Ito, Y., Sasaki, T.: Bioscience, biotechnology, and biochemistry cloning and characterization of the gene encoding a novel β-galactosidase from Bacillus Circulans. Biosci. Biotechnol. Biochem. 618, 1270–1276 (1997)CrossRefGoogle Scholar
  16. 16.
    Zeuner, B., Jers, C., Mikkelsen, J.D., Meyer, A.S.: Methods for improving enzymatic trans-glycosylation for synthesis of human milk oligosaccharide biomimetics. J. Agric. Food Chem. 62, 9615–9631 (2014)CrossRefPubMedGoogle Scholar
  17. 17.
    Sakai, K., Katsumi, R., Ohi, H., Usui, T., Ishido, Y.: Enzymatic syntheses of N-Acetyllactosamine and N-Acetylallolactosamine by the use of β-D-galactosidases. J. Carbohydr. Chem. 11, 553–565 (1992)CrossRefGoogle Scholar
  18. 18.
    Farkas, E., Thiem, J., Ajisaka, K.: Enzymatic synthesis of fucose-containing disaccharides employing the partially purified α-L-fucosidase from Penicillium multicolor. Carbohydr. Res. 328, 293–299 (2000)CrossRefPubMedGoogle Scholar
  19. 19.
    Bigge, J.C., Patel, T.P., Bruce, J.A., Goulding, P.N., Charles, S.M., Parekh, R.B.: Nonselective and efficient fluorescent labeling of glycans using 2-amino benzamide and anthranilic acid. Anal. Biochem. 230, 229–238 (1995)CrossRefPubMedGoogle Scholar
  20. 20.
    Mozaffar, Z., Nakanishi, K., Matsuno, R., Kamikubo, T.: Purification and properties of β-galactosidases from Bacillus circulans. Agric. Biol. Chem. 48, 3053–3061 (1984)Google Scholar
  21. 21.
    Kunz, C., Rudloff, S., Hintelmann, A., Pohlentz, G., Egge, H.: High-pH anion-exchange chromatography with pulsed amperometric detection and molar response factors of human milk oligosaccharides. J. Chromatogr. B Biomed. Appl. 685, 211–221 (1996)CrossRefPubMedGoogle Scholar
  22. 22.
    Borromei, C., Cavazza, A., Merusi, C., Corradini, C.: Characterization and quantitation of short-chain fructooligosaccharides and inulooligosaccharides in fermented milks by high-performance anion-exchange chromatography with pulsed amperometric detection. J. Sep. Sci. 32, 3635–3642 (2009)CrossRefPubMedGoogle Scholar
  23. 23.
    Corradini, C., Cavazza, A., Bignardi, C.: High-performance anion-exchange chromatography coupled with pulsed electrochemical detection as a powerful tool to evaluate carbohydrates of food interest: principles and applications. Int. J. Carbohydr. Chem. 2012, 1–13 (2012)CrossRefGoogle Scholar
  24. 24.
    Schwab, C., Lee, V., Sørensen, K.I., Gänzle, M.G.: Production of galactooligosaccharides and heterooligosaccharides with disrupted cell extracts and whole cells of lactic acid bacteria and bifidobacteria. Int. Dairy J. 21, 748–754 (2011)CrossRefGoogle Scholar
  25. 25.
    Black, B.A., Lee, V.S.Y., Zhao, Y.Y., Hu, Y., Curtis, J.M., Ga, M.G.: Structural identification of novel oligosaccharides produced by lactobacillus bulgaricus and lactobacillus plantarum. J. Agric. Food Chem. 60, 4886–4894 (2012)CrossRefPubMedGoogle Scholar
  26. 26.
    Van Leeuwen, S.S., Kuipers, B.J.H., Dijkhuizen, L., Kamerling, J.P.: 1H NMR analysis of the lactose/β-galactosidase-derived galacto-oligosaccharide components of Vivinal GOS up to DP5. Carbohydr. Res. 400, 59–73 (2014)CrossRefPubMedGoogle Scholar
  27. 27.
    Boehm, G., Stahl, B.: Oligosaccharides from milk. J. Nutr. 137, 847S–849S (2007)CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2017

Authors and Affiliations

  1. 1.Department of Bioproducts and BiosystemsAalto University School of Chemical EngineeringEspooFinland
  2. 2.Technical Research Center of FinlandEspooFinland
  3. 3.Valio Ltd.HelsinkiFinland

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