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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Biosynthesis of human milk oligosaccharides via metabolic engineering approaches: current advances and challenges

Current Opinion in Biotechnology, Journal Year: 2022, Volume and Issue: 78, P. 102841 - 102841

Published: Nov. 11, 2022

Language: Английский

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De novo biosynthesis of 2′-fucosyllactose in a metabolically engineered Escherichia coli using a novel ɑ1,2-fucosyltransferase from Azospirillum lipoferum

Bioresource Technology, Journal Year: 2023, Volume and Issue: 374, P. 128818 - 128818

Published: March 2, 2023

Language: Английский

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Elimination of Byproduct Generation and Enhancement of 2′-Fucosyllactose Synthesis by Expressing a Novel α1,2-Fucosyltransferase in Engineered Escherichia coli

Journal of Agricultural and Food Chemistry, Journal Year: 2023, Volume and Issue: 71(12), P. 4915 - 4923

Published: March 6, 2023

2′-Fucosyllactose (2′-FL) is a kind of fucosylated human milk oligosaccharide (HMO), representing the most abundant in breast milk. We conducted systematic studies on three canonical α1,2-fucosyltransferases (WbgL, FucT2, and WcfB) to quantify byproducts lacZ- wcaJ-deleted Escherichia coli BL21(DE3) basic host strain. Further, we screened highly active α1,2-fucosyltransferase from Helicobacter sp. 11S02629-2 (BKHT), which exhibits high vivo 2′-FL productivity without formation difucosyl lactose (DFL) 3-FL. The maximum titer yield reached 11.13 g/L 0.98 mol/mol lactose, respectively, shake-flask cultivation, both approaching theoretical value. In 5 L fed-batch 94.7 extracellularly with mol 2′-FL/mol 1.14 g L–1 h–1. Our reported highest date.

Language: Английский

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Engineering Escherichia coli for high-level production of lacto-N-fucopentaose I by stepwise de novo pathway construction

Carbohydrate Polymers, Journal Year: 2023, Volume and Issue: 315, P. 121028 - 121028

Published: May 15, 2023

Language: Английский

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Semi-rationally designed site-saturation mutation of Helicobacter pylori α-1,2-fucosyltransferase for improved catalytic activity and thermostability

International Journal of Biological Macromolecules, Journal Year: 2024, Volume and Issue: 259, P. 129316 - 129316

Published: Jan. 11, 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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De Novo Biosynthesis of Difucosyllactose by Artificial Pathway Construction and α1,3/4-Fucosyltransferase Rational Design in Escherichia coli

Journal of Agricultural and Food Chemistry, Journal Year: 2024, Volume and Issue: unknown

Published: April 10, 2024

Difucosyllactose (DFL) is a significant and plentiful oligosaccharide found in human breast milk. In this study, an artificial metabolic pathway of DFL was designed, focusing on the de novo biosynthesis GDP-fucose from only glycerol. This achieved by engineering

Language: Английский

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High-Level Production of l-Fucose by Plasmid-Free or Antibiotic-Independent Metabolically Engineered Escherichia coli Strains

ACS Sustainable Chemistry & Engineering, Journal Year: 2024, Volume and Issue: 12(17), P. 6619 - 6628

Published: April 17, 2024

l-Fucose is an important monosaccharide unit that exists in various biomasses, especially microalgae. Microbial l-fucose using metabolically engineered strains has attracted attention due to its high yield and industrial feasibility. Previously, we Escherichia coli MG1655 efficiently produce 2′-fucosyllactose by genomic integration. Herein, this plasmid-free strain was further integrating a specific α-l-fucosidase gene deleting the degradation pathway. Its effectiveness of biosynthesis inducer-free fermentation demonstrated both shake-flask fed-batch cultivation with titers 2.74 21.15 g/L, respectively. The precursor GDP-l-fucose supply strengthened obviously enhance introducing single plasmid expressing four pathway genes. hok/sok system introduced promote stabilization without antibiotic. final could antibiotics, 6.83 35.68 g/L fed-cultivation cultivation,

Language: Английский

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