Glycosyltransferases: glycoengineers in human milk oligosaccharide synthesis and manufacturing DOI Creative Commons

A. Slater,

Andrew G. McDonald, Rita M. Hickey

et al.

Frontiers in Molecular Biosciences, Journal Year: 2025, Volume and Issue: 12

Published: April 30, 2025

Human milk oligosaccharides (HMOs) are a diverse group of complex carbohydrates that play crucial roles in infant health, promoting beneficial gut microbiota, modulating immune responses, and protecting against pathogens. Central to the synthesis HMOs glycosyltransferases, specialized class enzymes catalyse transfer sugar moieties form glycan structures characteristic HMOs. This review provides an in-depth analysis beginning with their classification based on structural functional characteristics. The catalytic activity these is explored, highlighting mechanisms by which they facilitate precise addition monosaccharides HMO biosynthesis. Structural insights into glycosyltransferases also discussed, shedding light how conformational features enable specific glycosidic bond formations. maps out key biosynthetic pathways involved production, including lactose, subsequent fucosylation sialylation processes, all intricately regulated glycosyltransferases. Industrial methods for synthesis, chemical, enzymatic, microbial approaches, examined, emphasizing role processes. Finally, discusses future directions glycosyltransferase research, particularly enhancing efficiency developing advanced analytical techniques better understand complexity biological functions

Language: Английский

EASyMap-Guided Stepwise One-Pot Multienzyme (StOPMe) Synthesis and Multiplex Assays Identify Functional Tetraose-Core-Human Milk Oligosaccharides DOI Creative Commons
Yuanyuan Bai, Anand K. Agrahari, Libo Zhang

et al.

JACS Au, Journal Year: 2025, Volume and Issue: unknown

Published: Jan. 27, 2025

Carbohydrates are biologically and medicinally important molecules that attracting growing attention to their synthesis applications. Unlike the biosynthetic processes for nucleic acids proteins, carbohydrate biosynthesis is not template-driven, more challenging, often leads product variations. In lieu of templates biosynthesis, we describe herein a new concept designing enzyme assembly synthetic maps (EASyMaps) as blueprints guide glycosyltransferase-dependent stepwise one-pot multienzyme (StOPMe) systematically access structurally diverse carbohydrates in target-oriented manner. The strategy demonstrated construction comprehensive library tetraose-core-containing human milk oligosaccharides (HMOs) presenting functional glycan epitopes shared by complex HMOs. tetraose-core-HMOs attractive candidates large-scale production development HMO-based nutraceuticals. To achieve preparative-scale targets containing Neu5Acα2–6GlcNAc component, α2–6-sialyltransferase hST6GALNAC5 successfully expressed E. coli. Neoglycoproteins with controlled valencies prepared immobilized on fluorescent magnetic beads. Multiplex bead assays reveal ligands glycan-binding proteins from plants, influenza viruses, human, bacteria, identifying promising HMO EASyMaps StOPMe systematic manner broadly applicable beyond efficient process suitable can be potentially adapted automation.

Language: Английский

Citations

1
Research Progress on the Degradation of Human Milk Oligosaccharides (HMOs) by Bifidobacteria DOI Open Access

Ruitao Cai,

Jie Zhang, Yang Song

et al.

Nutrients, Journal Year: 2025, Volume and Issue: 17(3), P. 519 - 519

Published: Jan. 30, 2025

The purpose of this study was to investigate the degradation mechanism Bifidobacterium on breast milk oligosaccharides (HMOs) and its application in infant nutrition. composition characteristics HMOs were introduced, by described, including intracellular extracellular digestion species-specific differences. interaction between Bacteroides process degrading effect intestinal microecology analyzed. effects HMO formula powder microbiota infants discussed, simulating composition, regulating flora immune function, infection prevention, brain development. Finally, research results are summarized, future directions proposed provide for field

Language: Английский

Citations

0
Ketodeoxynonulosonic Acid Hydroxylase (Kdnase) Assisted Site‐Specific Enzymatic α2,6‐Sialylation DOI Open Access

Yu Zhou,

Yun Li, Jiayu Wen

et al.

Chinese Journal of Chemistry, Journal Year: 2025, Volume and Issue: unknown

Published: March 26, 2025

Comprehensive Summary Owing to its promiscuous substrate specificity and high catalytic efficiency, the bacterial α2,6‐sialyltransferase from Photobacterium damselae (Pd2,6ST) has been widely used for synthesis of various α2,6‐linked sialosides. However, Pd2,6ST is not a suitable enzyme regioselective α2,6‐sialylation complex acceptor substrates containing multiple galactose (Gal) and/or N ‐acetylgalactosamine (GalNAc) residues due specificity. In this study, novel enzymatic engineering strategy was developed overcome limitation by employing enzymatically introduced ketodeoxynonulosonic acid (Kdn) as temporary “protecting group” at unwanted sialylation sites. The Kdn can be selectively removed hydrolase Aspergillus fumigatus ( Af Kdnase) appropriate stage without affecting coexisting sialic residues, such ‐acetylneuraminic (Neu5Ac) or ‐glycolylneuraminic (Neu5Gc). This provides general practical approach sialosides, including sialylated poly‐LacNAc glycans, disialylated ganglioside glycan epitopes, branched human milk oligosaccharides.

Language: Английский

Citations

0
Glycosyltransferases: glycoengineers in human milk oligosaccharide synthesis and manufacturing DOI Creative Commons

A. Slater,

Andrew G. McDonald, Rita M. Hickey

et al.

Frontiers in Molecular Biosciences, Journal Year: 2025, Volume and Issue: 12

Published: April 30, 2025

Human milk oligosaccharides (HMOs) are a diverse group of complex carbohydrates that play crucial roles in infant health, promoting beneficial gut microbiota, modulating immune responses, and protecting against pathogens. Central to the synthesis HMOs glycosyltransferases, specialized class enzymes catalyse transfer sugar moieties form glycan structures characteristic HMOs. This review provides an in-depth analysis beginning with their classification based on structural functional characteristics. The catalytic activity these is explored, highlighting mechanisms by which they facilitate precise addition monosaccharides HMO biosynthesis. Structural insights into glycosyltransferases also discussed, shedding light how conformational features enable specific glycosidic bond formations. maps out key biosynthetic pathways involved production, including lactose, subsequent fucosylation sialylation processes, all intricately regulated glycosyltransferases. Industrial methods for synthesis, chemical, enzymatic, microbial approaches, examined, emphasizing role processes. Finally, discusses future directions glycosyltransferase research, particularly enhancing efficiency developing advanced analytical techniques better understand complexity biological functions

Language: Английский

Citations

0