High-Yield Synthesis of Lacto-N-Neotetraose from Glycerol and Glucose in Engineered Escherichia coli

Journal of Agricultural and Food Chemistry, Journal Year: 2024, Volume and Issue: 72(10), P. 5325 - 5338

Published: Jan. 26, 2024

Lacto-

Language: Английский

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Metabolic engineering of Escherichia coli BL21(DE3) cocultured with glucose and xylose for efficient production of 2′-fucosyllactose

Bioresource Technology, Journal Year: 2025, Volume and Issue: unknown, P. 132062 - 132062

Published: Jan. 1, 2025

Language: Английский

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Enzymatic modular synthesis of asymmetrically branched human milk oligosaccharides

Carbohydrate Polymers, Journal Year: 2024, Volume and Issue: 333, P. 121908 - 121908

Published: Feb. 9, 2024

Language: Английский

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EASyMap-Guided Stepwise One-Pot Multienzyme (StOPMe) Synthesis and Multiplex Assays Identify Functional Tetraose-Core-Human Milk Oligosaccharides

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

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Highly efficient biosynthesis of 3′-sialyllactose in engineered Escherichia coli

International Journal of Biological Macromolecules, Journal Year: 2024, Volume and Issue: 269, P. 132081 - 132081

Published: May 3, 2024

Language: Английский

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Microbial Synthesis of Lacto-N-fucopentaose I with High Titer and Purity by Screening of Specific Glycosyltransferase and Elimination of Residual Lacto-N-triose II and Lacto-N-tetraose

Journal of Agricultural and Food Chemistry, Journal Year: 2024, Volume and Issue: 72(8), P. 4317 - 4324

Published: Feb. 16, 2024

Lacto-N-fucopentaose I (LNFP I) has recently been approved as generally recognized safe, demonstrating its great commercial potential in the food industry. Microbial synthesis through metabolic engineering strategies is an effective approach for large-scale production of LNFP I. Biosynthesis requires consideration two key points: high titer with low byproduct 2′-fucosyllactose (2′-FL) generation and purity lacto-N-triose II (LNTri II) lacto-N-tetraose (LNT) residues. Herein, α1,2-fucosyltransferase from Thermoanaerobacterium sp. RBIITD was screened 16 selected I-producing glycosyltransferase candidates, showing highest vivo productivity. Chromosomal integration wbgO enhanced by improving precursor conversion LNTri to LNT. The best engineered strain produced 4.42 35.1 g/L shake-flask fed-batch cultivation, respectively. residual LNT were eliminated further cultivation a recombinant coexpressing Bifidobacterium bifidum β-N-acetylhexosaminidase lacto-N-biosidase. A strategy biosynthesis yield finally realized, providing support practical application production.

Language: Английский

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Highly-efficient in vivo production of lacto-N-fucopentaose V by a regio-specific α1,3/4-fucosyltransferase from Bacteroides fragilis NCTC 9343

International Journal of Biological Macromolecules, Journal Year: 2024, Volume and Issue: 266, P. 130955 - 130955

Published: March 16, 2024

Language: Английский

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Microbial modifications with Lycium barbarum L. oligosaccharides decrease hepatic fibrosis and mitochondrial abnormalities in mice

Phytomedicine, Journal Year: 2023, Volume and Issue: 120, P. 155068 - 155068

Published: Sept. 4, 2023

Language: Английский

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An Overview of Sugar Nucleotide-Dependent Glycosyltransferases for Human Milk Oligosaccharide Synthesis

Journal of Agricultural and Food Chemistry, Journal Year: 2023, Volume and Issue: 71(33), P. 12390 - 12402

Published: Aug. 8, 2023

Human milk oligosaccharides (HMOs) have received increasing attention because of their special effects on infant health and commercial value as the new generation core components in formula. Currently, large-scale production HMOs is generally based microbial synthesis using metabolically engineered cell factories. Introduction specific glycosyltransferases essential for construction HMO-producing strains which are sugar nucleotide-dependent. Four types been used typical glycosylation reactions to synthesize HMOs. Soluble expression, substrate specificity, regioselectivity common concerns these practical applications. Screening an important research topic solve problems. Molecular modification has also performed enhance catalytic activity various improve specificity regioselectivity. In this article, nucleotide-dependent HMO were overviewed, described, future perspectives glycosyltransferase-related studies provided.

Language: Английский

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Efficient fermentative production of lactodifucotetraose by controlling sequential glycosyltransferase reactions in Escherichia coli

Biotechnology Progress, Journal Year: 2025, Volume and Issue: unknown

Published: Feb. 6, 2025

Abstract Lactodifucotetraose (LDFT) is a human milk oligosaccharide (HMO) that might reduce inflammation in infants. In this study, we established useful production process of LDFT by engineering two key enzymes, α1,2‐fucosyltransferase (α1,2‐FucT) and α1,3‐fucosyltransferase (α1,3‐FucT). First, verified which 2′‐fucosyllactose (2′‐FL) or 3‐fucosyllactose (3‐FL) (mostly unverified) was more useful. We searched for FucTs functioned efficiently vivo against the raw material lactose intermediates 2′‐FL 3‐FL external substrate addition to culture medium. found α1,2‐ FucT (HMFT) from Helicobacter mustelae N‐terminal truncated form α1,3‐FucT Bacteroides fragilis (BfFucTΔN10) had high potential. not converted LDFT, be attributed low reactivity HMFT as well uptake efficiency LacY, revealed growth test with exogenously added FL sole carbon source heterologously expressed intracellular fucosidase. Furthermore, because accumulation negative impact on cell growth, avoided route passing through 3‐FL. By adjusting copy numbers BffucTΔN10 , produced predominantly via 2′‐FL. Finally, 17.5 g/L (with 6.8 no residual lactose) accumulated 3‐L fed‐batch after 77 h. This study reports detailed analysis multiple pathways shows control glycosyltransferases can improve complex HMOs.

Language: Английский

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