Synthesis of human milk oligosaccharides: Protein engineering strategies for improved enzymatic transglycosylation

Birgitte Zeuner, David Teze, Jan Muschiol, Anne S. Meyer*

*Corresponding author for this work

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Abstract

Human milk oligosaccharides (HMOs) signify a unique group of oligosaccharides in breast milk, which is of major importance for infant health and development. The functional benefits of HMOs create an enormous impetus for biosynthetic production of HMOs for use as additives in infant formula and other products. HMO molecules can be synthesized chemically, via fermentation, and by enzymatic synthesis. This treatise discusses these different techniques, with particular focus on harnessing enzymes for controlled enzymatic synthesis of HMO molecules. In order to foster precise and high-yield enzymatic synthesis, several novel protein engineering approaches have been reported, mainly concerning changing glycoside hydrolases to catalyze relevant transglycosylations. The protein engineering strategies for these enzymes range from rationally modifying specific catalytic residues, over targeted subsite -1 mutations, to unique and novel transplantations of designed peptide sequences near the active site, so-called loop engineering. These strategies have proven useful to foster enhanced transglycosylation to promote different types of HMO synthesis reactions. The rationale of subsite -1 modification, acceptor binding site matching, and loop engineering, including changes that may alter the spatial arrangement of water in the enzyme active site region, may prove useful for novel enzyme-catalyzed carbohydrate design in general.

Original languageEnglish
Article number2033
JournalMolecules
Volume24
Issue number11
Number of pages22
ISSN1420-3049
DOIs
Publication statusPublished - 2019

Keywords

  • Casein glycomacropeptide
  • Fucosidase
  • Human milk oligosaccharides
  • Protein engineering
  • Sialidase
  • Transfucosylation
  • Transglycosylation
  • Transsialylation
  • β-N-acetylhexosaminidase

Cite this

@article{cd550e346a8b4e1cb482973a48811bb3,
title = "Synthesis of human milk oligosaccharides: Protein engineering strategies for improved enzymatic transglycosylation",
abstract = "Human milk oligosaccharides (HMOs) signify a unique group of oligosaccharides in breast milk, which is of major importance for infant health and development. The functional benefits of HMOs create an enormous impetus for biosynthetic production of HMOs for use as additives in infant formula and other products. HMO molecules can be synthesized chemically, via fermentation, and by enzymatic synthesis. This treatise discusses these different techniques, with particular focus on harnessing enzymes for controlled enzymatic synthesis of HMO molecules. In order to foster precise and high-yield enzymatic synthesis, several novel protein engineering approaches have been reported, mainly concerning changing glycoside hydrolases to catalyze relevant transglycosylations. The protein engineering strategies for these enzymes range from rationally modifying specific catalytic residues, over targeted subsite -1 mutations, to unique and novel transplantations of designed peptide sequences near the active site, so-called loop engineering. These strategies have proven useful to foster enhanced transglycosylation to promote different types of HMO synthesis reactions. The rationale of subsite -1 modification, acceptor binding site matching, and loop engineering, including changes that may alter the spatial arrangement of water in the enzyme active site region, may prove useful for novel enzyme-catalyzed carbohydrate design in general.",
keywords = "Casein glycomacropeptide, Fucosidase, Human milk oligosaccharides, Protein engineering, Sialidase, Transfucosylation, Transglycosylation, Transsialylation, β-N-acetylhexosaminidase",
author = "Birgitte Zeuner and David Teze and Jan Muschiol and Meyer, {Anne S.}",
year = "2019",
doi = "10.3390/molecules24112033",
language = "English",
volume = "24",
journal = "Molecules",
issn = "1420-3049",
publisher = "M D P I AG",
number = "11",

}

TY - JOUR

T1 - Synthesis of human milk oligosaccharides: Protein engineering strategies for improved enzymatic transglycosylation

AU - Zeuner, Birgitte

AU - Teze, David

AU - Muschiol, Jan

AU - Meyer, Anne S.

PY - 2019

Y1 - 2019

N2 - Human milk oligosaccharides (HMOs) signify a unique group of oligosaccharides in breast milk, which is of major importance for infant health and development. The functional benefits of HMOs create an enormous impetus for biosynthetic production of HMOs for use as additives in infant formula and other products. HMO molecules can be synthesized chemically, via fermentation, and by enzymatic synthesis. This treatise discusses these different techniques, with particular focus on harnessing enzymes for controlled enzymatic synthesis of HMO molecules. In order to foster precise and high-yield enzymatic synthesis, several novel protein engineering approaches have been reported, mainly concerning changing glycoside hydrolases to catalyze relevant transglycosylations. The protein engineering strategies for these enzymes range from rationally modifying specific catalytic residues, over targeted subsite -1 mutations, to unique and novel transplantations of designed peptide sequences near the active site, so-called loop engineering. These strategies have proven useful to foster enhanced transglycosylation to promote different types of HMO synthesis reactions. The rationale of subsite -1 modification, acceptor binding site matching, and loop engineering, including changes that may alter the spatial arrangement of water in the enzyme active site region, may prove useful for novel enzyme-catalyzed carbohydrate design in general.

AB - Human milk oligosaccharides (HMOs) signify a unique group of oligosaccharides in breast milk, which is of major importance for infant health and development. The functional benefits of HMOs create an enormous impetus for biosynthetic production of HMOs for use as additives in infant formula and other products. HMO molecules can be synthesized chemically, via fermentation, and by enzymatic synthesis. This treatise discusses these different techniques, with particular focus on harnessing enzymes for controlled enzymatic synthesis of HMO molecules. In order to foster precise and high-yield enzymatic synthesis, several novel protein engineering approaches have been reported, mainly concerning changing glycoside hydrolases to catalyze relevant transglycosylations. The protein engineering strategies for these enzymes range from rationally modifying specific catalytic residues, over targeted subsite -1 mutations, to unique and novel transplantations of designed peptide sequences near the active site, so-called loop engineering. These strategies have proven useful to foster enhanced transglycosylation to promote different types of HMO synthesis reactions. The rationale of subsite -1 modification, acceptor binding site matching, and loop engineering, including changes that may alter the spatial arrangement of water in the enzyme active site region, may prove useful for novel enzyme-catalyzed carbohydrate design in general.

KW - Casein glycomacropeptide

KW - Fucosidase

KW - Human milk oligosaccharides

KW - Protein engineering

KW - Sialidase

KW - Transfucosylation

KW - Transglycosylation

KW - Transsialylation

KW - β-N-acetylhexosaminidase

U2 - 10.3390/molecules24112033

DO - 10.3390/molecules24112033

M3 - Review

VL - 24

JO - Molecules

JF - Molecules

SN - 1420-3049

IS - 11

M1 - 2033

ER -