A chemo-enzymatic approach for the synthesis of human milk oligosaccharide backbone structures

Jan Muschiol*, Anne S. Meyer

*Corresponding author for this work

Research output: Contribution to journalJournal articleResearchpeer-review

Abstract

The ability of an engineered β-N-acetylhexosaminidase to utilize a reactive oxazoline as donor molecule for transglycosylation reaction to synthesize human milk oligosaccharide backbone structures was studied. The human milk oligosaccharide precursor lacto-N-triose II and three regioisomers could be synthesized using the oxazoline, which was either in situ-generated resulting in a chemo-enzymatic sequential cascade or was used as a purified compound. The highest observed concentration of overall transglycosylation products in a cascade reaction was 13.7 mM after 18.5 h, whereas the use of purified oxazoline resulted in 25.0 mM of transglycosylation products after 6.5 h. Remarkably, the in situ-generated oxazoline could be used without any further purification and it was shown that the used enzyme tolerated significant amounts of reagents such as triethylamine, which is reported for the first time for an enzyme from the glycoside hydrolase family 20.

Original languageEnglish
JournalZeitschrift fuer Naturforschung. Section C: A Journal of Biosciences
Volume74
Issue number3-4
Pages (from-to)85-89
Number of pages5
ISSN0939-5075
DOIs
Publication statusPublished - 2019

Keywords

  • β-N-acetylhexosaminidase
  • Chemo-enzymatic cascade
  • Human milk oligosaccharides
  • Lacto-N-triose II
  • Transglycosylation

Cite this

@article{e71418c0fb99427a863a26be0fbe471c,
title = "A chemo-enzymatic approach for the synthesis of human milk oligosaccharide backbone structures",
abstract = "The ability of an engineered β-N-acetylhexosaminidase to utilize a reactive oxazoline as donor molecule for transglycosylation reaction to synthesize human milk oligosaccharide backbone structures was studied. The human milk oligosaccharide precursor lacto-N-triose II and three regioisomers could be synthesized using the oxazoline, which was either in situ-generated resulting in a chemo-enzymatic sequential cascade or was used as a purified compound. The highest observed concentration of overall transglycosylation products in a cascade reaction was 13.7 mM after 18.5 h, whereas the use of purified oxazoline resulted in 25.0 mM of transglycosylation products after 6.5 h. Remarkably, the in situ-generated oxazoline could be used without any further purification and it was shown that the used enzyme tolerated significant amounts of reagents such as triethylamine, which is reported for the first time for an enzyme from the glycoside hydrolase family 20.",
keywords = "β-N-acetylhexosaminidase, Chemo-enzymatic cascade, Human milk oligosaccharides, Lacto-N-triose II, Transglycosylation",
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year = "2019",
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language = "English",
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journal = "Zeitschrift fuer Naturforschung. Section C: A Journal of Biosciences",
issn = "0939-5075",
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A chemo-enzymatic approach for the synthesis of human milk oligosaccharide backbone structures. / Muschiol, Jan; Meyer, Anne S.

In: Zeitschrift fuer Naturforschung. Section C: A Journal of Biosciences, Vol. 74, No. 3-4, 2019, p. 85-89.

Research output: Contribution to journalJournal articleResearchpeer-review

TY - JOUR

T1 - A chemo-enzymatic approach for the synthesis of human milk oligosaccharide backbone structures

AU - Muschiol, Jan

AU - Meyer, Anne S.

PY - 2019

Y1 - 2019

N2 - The ability of an engineered β-N-acetylhexosaminidase to utilize a reactive oxazoline as donor molecule for transglycosylation reaction to synthesize human milk oligosaccharide backbone structures was studied. The human milk oligosaccharide precursor lacto-N-triose II and three regioisomers could be synthesized using the oxazoline, which was either in situ-generated resulting in a chemo-enzymatic sequential cascade or was used as a purified compound. The highest observed concentration of overall transglycosylation products in a cascade reaction was 13.7 mM after 18.5 h, whereas the use of purified oxazoline resulted in 25.0 mM of transglycosylation products after 6.5 h. Remarkably, the in situ-generated oxazoline could be used without any further purification and it was shown that the used enzyme tolerated significant amounts of reagents such as triethylamine, which is reported for the first time for an enzyme from the glycoside hydrolase family 20.

AB - The ability of an engineered β-N-acetylhexosaminidase to utilize a reactive oxazoline as donor molecule for transglycosylation reaction to synthesize human milk oligosaccharide backbone structures was studied. The human milk oligosaccharide precursor lacto-N-triose II and three regioisomers could be synthesized using the oxazoline, which was either in situ-generated resulting in a chemo-enzymatic sequential cascade or was used as a purified compound. The highest observed concentration of overall transglycosylation products in a cascade reaction was 13.7 mM after 18.5 h, whereas the use of purified oxazoline resulted in 25.0 mM of transglycosylation products after 6.5 h. Remarkably, the in situ-generated oxazoline could be used without any further purification and it was shown that the used enzyme tolerated significant amounts of reagents such as triethylamine, which is reported for the first time for an enzyme from the glycoside hydrolase family 20.

KW - β-N-acetylhexosaminidase

KW - Chemo-enzymatic cascade

KW - Human milk oligosaccharides

KW - Lacto-N-triose II

KW - Transglycosylation

U2 - 10.1515/znc-2018-0149

DO - 10.1515/znc-2018-0149

M3 - Journal article

VL - 74

SP - 85

EP - 89

JO - Zeitschrift fuer Naturforschung. Section C: A Journal of Biosciences

JF - Zeitschrift fuer Naturforschung. Section C: A Journal of Biosciences

SN - 0939-5075

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ER -