Highly Efficient In Vivo Production of Sialyllacto-N-tetraose C via Screening of Beneficial β1,4-galactosyltransferase and α2,6-sialyltransferase

Journal of Agricultural and Food Chemistry, Journal Year: 2025, Volume and Issue: 73(9), P. 5376 - 5384

Published: Feb. 18, 2025

Biological production of human milk oligosaccharides (HMOs) using metabolically engineered strains is a research hotspot in food biotechnology, but less effort has been made on the biological sialylated complex HMOs. Sialyllacto-N-tetraose c only monosialylated HMO top 15 In this study, metabolic pathway LST was constructed Escherichia coli BL21(DE3) by introducing three sequential glycosyltransferases: β1,3-N-acetylglucosaminyltransferase, β1,4-galactosyltransferase, and α2,6-sialyltransferase. The cytidine 5′-monophospho (CMP)-N-acetylneuraminic acid (Neu5Ac) enhanced to improve production. β1,4-galactosyltransferase from Helicobacter pylori J99 (HpGalT) α2,6-sialyltransferase Vespertiliibacter pulmonis (ED6ST) were screened as pair key glycosyltransferases for enhancing final strain could produce 1.718 9.745 g/L shake-flask fed-batch cultivation, respectively, indicating feasibility efficient biosynthesis

Language: Английский

---

Recent trends in human milk oligosaccharides: New synthesis technology, regulatory effects, and mechanisms of non‐intestinal functions

Comprehensive Reviews in Food Science and Food Safety, Journal Year: 2025, Volume and Issue: 24(2)

Published: March 1, 2025

Abstract Recently, the non‐intestinal functions of human milk oligosaccharides (HMOs) have been widely documented, including their roles in promoting brain development and growth, as well ameliorating anxiety, allergies, obesity. Understanding mechanisms action is becoming increasingly critical. Furthermore, these effects are frequently associated with type structure HMOs. As an innovative technology, “plant factory” expected to complement traditional synthesis technology. This study reviews novel techniques. Particular emphasis placed on processes, advantages, limitations technology can express genes related HMO instantaneously plant leaves, thereby enabling rapid cost‐effective generation However, remains underdeveloped, challenges low yield unsustainable production must be addressed. we present overview most recent clinical preclinical studies review emphasizes underlying HMOs primarily exert through cleavage beneficial monomer components, metabolism produce advantageous metabolites, regulation immune responses.

Language: Английский

---

Structure, Function, Synthesis and Improved Strategies of Fucosylated Human Milk Oligosaccharides and Their Future Perspectives: A Review

Food Bioscience, Journal Year: 2025, Volume and Issue: unknown, P. 106584 - 106584

Published: April 1, 2025

Language: Английский

---

Investigation of mannitol as a potential substrate for production of 2′-fucosyllactose in Yarrowia lipolytica

Bioresource Technology, Journal Year: 2025, Volume and Issue: 430, P. 132583 - 132583

Published: April 23, 2025

Language: Английский

---

Glycosyltransferases in human milk oligosaccharide synthesis: structural mechanisms and rational design

Current Opinion in Biotechnology, Journal Year: 2025, Volume and Issue: 93, P. 103315 - 103315

Published: May 9, 2025

Language: Английский

---

More than nutrition: Therapeutic potential and mechanism of human milk oligosaccharides against necrotizing enterocolitis

Life Sciences, Journal Year: 2024, Volume and Issue: 339, P. 122420 - 122420

Published: Jan. 11, 2024

Language: Английский

---

Gut Microbial Sialidases and Their Role in the Metabolism of Human Milk Sialylated Glycans

International Journal of Molecular Sciences, Journal Year: 2023, Volume and Issue: 24(12), P. 9994 - 9994

Published: June 10, 2023

Sialic acids (SAs) are α-keto-acid sugars with a nine-carbon backbone present at the non-reducing end of human milk oligosaccharides and glycan moiety glycoconjugates. SAs displayed on cell surfaces participate in regulation many physiologically important cellular molecular processes, including signaling adhesion. Additionally, sialyl-oligosaccharides from act as prebiotics colon by promoting settling proliferation specific bacteria SA metabolism capabilities. Sialidases glycosyl hydrolases that release α-2,3-, α-2,6- α-2,8-glycosidic linkages terminal residues oligosaccharides, glycoproteins glycolipids. The research sialidases has been traditionally focused pathogenic microorganisms, where these enzymes considered virulence factors. There is now growing interest commensal probiotic their potential transglycosylation activity for production functional mimics to complement infant formulas. This review provides an overview exo-alpha-sialidases gastrointestinal tract some insights into biological role biotechnological applications.

Language: Английский

---

Efficient Biosynthesis of Lacto-N-Biose I, a Building Block of Type I Human Milk Oligosaccharides, by a Metabolically Engineered Escherichia coli

Journal of Agricultural and Food Chemistry, Journal Year: 2024, Volume and Issue: 72(11), P. 5860 - 5866

Published: March 7, 2024

Lacto-N-biose I (LNB), termed a Type 1 disaccharide, is an important building block of human milk oligosaccharides. It shows promising prebiotic activity by stimulating the proliferation many gut-associated bifidobacteria and thus displays good potential in infant foods or supplements. Enzymatic microbial approaches to LNB synthesis have been studied, almost all which involve glycosylation phosphorylase as final step. Herein, we report new easier strategy through route "lactose → lacto-N-triose II (LNTri II) lacto-N-tetraose (LNT) LNB". A previously constructed LNT-producing Escherichia coli BL21(DE3) strain was engineered for biosynthesis introducing Bifidobacterium bifidum LnbB. efficiently produced, accompanied lactose regeneration. Genomic integration key pathway genes related LNTri LNT performed enhance titers. The produced 3.54 26.88 g/L shake-flask fed-batch cultivation, respectively.

Language: Английский

---

Engineered β1-3-N-Acetylglucosaminyltransferase Facilitating the One-Pot Multienzyme Synthesis of Human Milk Oligosaccharides

Journal of Agricultural and Food Chemistry, Journal Year: 2024, Volume and Issue: 72(50), P. 28019 - 28027

Published: Dec. 6, 2024

β1-3-linked N-acetylglucosaminide is a prevalent carbohydrate motif found in oligosaccharides, polysaccharides, glycoproteins, and glycolipids. It crucial component of human milk oligosaccharides (HMOs). Neisseria meningitidis β1-3-N-acetylglucosaminyltransferase (NmLgtA) catalyzes the formation glycosidic bond has potential for use synthesizing HMOs. However, this application hindered by challenges such as low levels enzyme expression, poor stability, significant aggregation. Since there no available crystal structure NmLgtA, we used its AlphaFold 2 predicted to identify unfavorable factors. We then modified removing 17 N-terminal amino acids substituting nine specific residues. The engineered NmLgtA-Opti exhibited improved thermal increased soluble protein complete relief from aggregation, enhanced catalysis while maintaining catalytic specificity substrate promiscuity. Furthermore, maximizes utilization can be employed sequential one-pot multienzyme platform high-yield production

Language: Английский

---

Construction of a plasmid-free Escherichia coli strain for lacto-N-neotetraose biosynthesis

Systems Microbiology and Biomanufacturing, Journal Year: 2024, Volume and Issue: 4(3), P. 965 - 982

Published: March 18, 2024

Language: Английский

---