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

Bioresource Technology, Год журнала: 2025, Номер unknown, С. 132062 - 132062

Опубликована: Янв. 1, 2025

Язык: Английский

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

Current Opinion in Biotechnology, Год журнала: 2022, Номер 78, С. 102841 - 102841

Опубликована: Ноя. 11, 2022

Язык: 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, Год журнала: 2023, Номер 374, С. 128818 - 128818

Опубликована: Март 2, 2023

Язык: 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, Год журнала: 2023, Номер 71(12), С. 4915 - 4923

Опубликована: Март 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.

Язык: 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, Год журнала: 2024, Номер 259, С. 129316 - 129316

Опубликована: Янв. 11, 2024

Язык: 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, Год журнала: 2023, Номер 315, С. 121028 - 121028

Опубликована: Май 15, 2023

Язык: Английский

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Engineering Escherichia coli MG1655 for Highly Efficient Biosynthesis of 2′-Fucosyllactose by De Novo GDP-Fucose Pathway

Journal of Agricultural and Food Chemistry, Год журнала: 2023, Номер 71(40), С. 14678 - 14686

Опубликована: Сен. 29, 2023

2'-Fucosyllactose (2'-FL), the most typical human milk oligosaccharide, is used as an additive in premium infant formula. Herein, we constructed two highly effective 2'-FL synthesis producers via a de novo GDP-fucose pathway from engineered Escherichia coli MG1655. First, lacZ and wcaJ, competitive genes, were disrupted to block invalid consumption of lactose GDP-fucose, respectively. Next, lacY gene was strengthened by switching its native promoter PJ23119. To enhance supply endogenous promoters clusters manC-manB gmd-fcl individually or combination. Subsequently, chromosomal integration constitutive PJ23119 promoter-based BKHT expression cassette (PJ23119-BKHT) performed arsB recA loci. The productive plasmid-based plasmid-free strains produced 76.9 50.1 g/L fed-batch cultivation, Neither them generated difucosyl nor 3-fucosyllactose byproducts.

Язык: 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, Год журнала: 2024, Номер 72(8), С. 4317 - 4324

Опубликована: Фев. 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.

Язык: 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, Год журнала: 2024, Номер 266, С. 130955 - 130955

Опубликована: Март 16, 2024

Язык: 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, Год журнала: 2024, Номер unknown

Опубликована: Апрель 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

Язык: Английский

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