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Article

Enzymatic Cascades for Tailored 13C6 and 15N Enriched Human Milk Oligosaccharides

1
Laboratory for Biomaterials, Institute for Biotechnology and Helmholtz-Institute for Biomedical Engineering, RWTH Aachen University, Pauwelsstraße 20, 52074 Aachen, Germany
2
Max Planck Institute for Dynamics of Complex Technical Systems, Sandtorstraße 1, 39106 Magdeburg, Germany
3
Institute of Functional Interfaces, Karlsruhe Institute of Technology, 76131 Karlsruhe, Germany
4
Chair of Bioprocess Engineering, Otto-von-Guericke-University, Universitätsplatz 2, 39106 Magdeburg, Germany
5
glyXera GmbH, Leipziger Straße 44, 39120 Magdeburg, Germany
*
Authors to whom correspondence should be addressed.
These authors contributed equally to this work.
Molecules 2019, 24(19), 3482; https://doi.org/10.3390/molecules24193482
Received: 16 July 2019 / Revised: 6 September 2019 / Accepted: 22 September 2019 / Published: 25 September 2019
(This article belongs to the Section Bioorganic Chemistry)
Several health benefits, associated with human milk oligosaccharides (HMOS), have been revealed in the last decades. Further progress, however, requires not only the establishment of a simple “routine” method for absolute quantification of complex HMOS mixtures but also the development of novel synthesis strategies to improve access to tailored HMOS. Here, we introduce a combination of salvage-like nucleotide sugar-producing enzyme cascades with Leloir-glycosyltransferases in a sequential pattern for the convenient tailoring of stable isotope-labeled HMOS. We demonstrate the assembly of [13C6]galactose into lacto-N- and lacto-N-neo-type HMOS structures up to octaoses. Further, we present the enzymatic production of UDP-[15N]GlcNAc and its application for the enzymatic synthesis of [13C6/15N]lacto-N-neo-tetraose for the first time. An exemplary application was selected—analysis of tetraose in complex biological mixtures—to show the potential of tailored stable isotope reference standards for the mass spectrometry-based quantification, using matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF-MS) as a fast and straightforward method for absolute quantification of HMOS. Together with the newly available well-defined tailored isotopic HMOS, this can make a crucial contribution to prospective research aiming for a more profound understanding of HMOS structure-function relations. View Full-Text
Keywords: glycobiology; biocatalysis; human milk oligosaccharides; isotopic labeling; quantification: nucleotide sugars; lacto-N-biose type; lacto-N-neo type; glycosyltransferases; capillary gel electrophoresis; MALDI-MS glycobiology; biocatalysis; human milk oligosaccharides; isotopic labeling; quantification: nucleotide sugars; lacto-N-biose type; lacto-N-neo type; glycosyltransferases; capillary gel electrophoresis; MALDI-MS
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MDPI and ACS Style

Fischöder, T.; Cajic, S.; Grote, V.; Heinzler, R.; Reichl, U.; Franzreb, M.; Rapp, E.; Elling, L. Enzymatic Cascades for Tailored 13C6 and 15N Enriched Human Milk Oligosaccharides. Molecules 2019, 24, 3482. https://doi.org/10.3390/molecules24193482

AMA Style

Fischöder T, Cajic S, Grote V, Heinzler R, Reichl U, Franzreb M, Rapp E, Elling L. Enzymatic Cascades for Tailored 13C6 and 15N Enriched Human Milk Oligosaccharides. Molecules. 2019; 24(19):3482. https://doi.org/10.3390/molecules24193482

Chicago/Turabian Style

Fischöder, Thomas, Samanta Cajic, Valerian Grote, Raphael Heinzler, Udo Reichl, Matthias Franzreb, Erdmann Rapp, and Lothar Elling. 2019. "Enzymatic Cascades for Tailored 13C6 and 15N Enriched Human Milk Oligosaccharides" Molecules 24, no. 19: 3482. https://doi.org/10.3390/molecules24193482

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