Combinatorial Optimization Strategies for 3-Fucosyllactose Hyperproduction in Escherichia coli

Journal of Agricultural and Food Chemistry, Journal Year: 2024, Volume and Issue: 72(25), P. 14191 - 14198

Published: June 15, 2024

3-Fucosyllactose (3-FL), an important fucosylated human milk oligosaccharide in breast milk, offers numerous health benefits to infants. Previously, we metabolically engineered

Language: Английский

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Microbial Synthesis of l-Fucose with High Productivity by a Metabolically Engineered Escherichia coli

Journal of Agricultural and Food Chemistry, Journal Year: 2023, Volume and Issue: 71(5), P. 2464 - 2471

Published: Jan. 26, 2023

l-Fucose is a natural deoxy hexose found in variety of organisms. It possesses many physiological effects and has potential applications pharmaceutical, cosmetic, food industries. Microbial synthesis via metabolic engineering attracts increasing attention for efficient production important chemicals. Previously, we reported the construction metabolically engineered Escherichia coli strain with high 2'-fucosyllactose productivity. Herein, further introduced Bifidobacterium bifidum α-l-fucosidase both plasmid expression genomic integration blocked l-fucose assimilation pathway by deleting fucI, fucK, rhaA. The highest titers reached 6.31 51.05 g/L shake-flask fed-batch cultivation, respectively. was little affected lactose added, there almost no residue throughout cultivation processes. productivity 0.76 g/L/h, indicating significant large-scale industrial applications.

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

Journal of Agricultural and Food Chemistry, Journal Year: 2023, Volume and Issue: 71(40), P. 14678 - 14686

Published: Sept. 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.

Language: Английский

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Metabolic Engineering of Escherichia coli BL21(DE3) for 2′-Fucosyllactose Synthesis in a Higher Productivity

ACS Synthetic Biology, Journal Year: 2025, Volume and Issue: unknown

Published: Jan. 15, 2025

2′-Fucosyllactose (2′-FL) is the most abundant human milk oligosaccharides (HMOs). 2′-FL exhibits great benefits for infant health, such as preventing infantile diarrhea and promoting growth of intestinal probiotics. The microbial cell factory technique has shown promise massive production 2′-FL. Here, we aimed to construct a recombinant E. coli BL21(DE3) strain hyperproduction Initially, multicopy genomic integration expression lactose permease gene lacY reduced formation byproducts. Furthermore, more efficient Shine–Dalgarno sequence was used replace wild-type in manC-manB gmd-wcaG clusters, which significantly increased titer. Based on these results, overexpressed sugar efflux transporter SetA knocked out pgi gene. This further improved synthesis when glycerol sole carbon source. Finally, new α-1,2-fucosyltransferase identified Neisseria sp., exhibited higher capacity production. Fed-batch fermentation produced 141.27 g/L 45 h with productivity 3.14 × h. rate achieved highest recorded levels, indicating potential engineered BL21 (DE3) strains use industrial

Language: Английский

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Biosynthesis and metabolic engineering of natural sweeteners

AMB Express, Journal Year: 2025, Volume and Issue: 15(1)

Published: March 18, 2025

Natural sweeteners have attracted widespread attention because they are eco-friendly, healthy, low in calories, and tasty. The demand for natural is increasing together with the popularity of green, low-carbon, sustainable development. With development synthetic biology, microbial cell factories emerged as an effective method to produce large amounts sweeteners. This technology has significantly progressed recent years. review summarizes pathways enzymes related biosynthesis sweeteners, such mogrosides, steviol glycosides, glycyrrhizin, glycyrrhetinic acid, phlorizin, trilobatin, erythritol, sorbitol, mannitol, thaumatin, monellin, brazzein. Moreover, it focuses on research about production these using biology methods, aiming provide a reference future

Language: Английский

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Dual Antibiotic-Free Plasmid Systems Enable High-Efficiency l-Fucose Biosynthesis

ACS Synthetic Biology, Journal Year: 2025, Volume and Issue: unknown

Published: March 27, 2025

l-Fucose, a functional monosaccharide with significant commercial potential in the pharmaceutical, nutraceutical, and cosmetic industries, faces challenges microbial production due to antibiotic-dependent plasmid maintenance systems. This study presents dual antibiotic-free strategy engineered Escherichia coli BL21(DE3) achieve high-efficiency l-fucose biosynthesis. By integration of hok/sok toxin-antitoxin system cysC-based auxotrophic selection into two plasmids, genetic stability retention were ensured without antibiotics. Metabolic pathway optimization involved enhancing GDP-l-fucose supply via promoter replacements, genomic key enzymes (α1,2-fucosyltransferase α-l-fucosidase), blocking degradation. The strain demonstrated robust performance, producing 7.99 g/L shake-flask fermentation 61.91 fed-batch cultivation─both antibiotic-free. titer represents highest reported yield date, highlighting effectiveness combining toxin–antitoxin systems for sustainable, high-productivity manufacturing.

Language: Английский

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Supplementation of 2′-Fucosyllactose during the Growth Period Improves Neurodevelopmental Disorders in Offspring Mice Induced by Maternal Immune Activation

Journal of Agricultural and Food Chemistry, Journal Year: 2025, Volume and Issue: unknown

Published: May 11, 2025

Autism spectrum disorder is a serious neurodevelopmental whose early onset significantly affects an individual's social interactions and cognitive function. Recent research suggests that modulating the gut microbiota could be potential intervention strategy for autism symptoms. 2'-Fucosyllactose has been identified as regulator of homeostasis, however, its effectiveness in addressing remains unclear. In this study, effects daily supplementation 2'-FL 3-week-old male offspring mice 5 weeks were examined. The results showed 2'-fucosyllactose improved disorder-like behavioral deficits. Furthermore, with restored intestinal barrier integrity increased relative abundance beneficial bacteria, particularly Akkermansia Bifidobacterium are closely related to bile acid metabolism. Notably, treatment elevated content acids upregulated expression receptors brain. Co-housing experiments further confirmed crucial role mediating 2'-fucosyllactose. Overall, study alleviate maternal immune activation-induced deficits neuroinflammation through regulation gut-brain axis, offering therapeutic value.

Language: Английский

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Recent advances in a functional deoxy hexose l-fucose: Occurrence, physiological effects, and preparation

Trends in Food Science & Technology, Journal Year: 2023, Volume and Issue: 138, P. 16 - 26

Published: May 29, 2023

Language: Английский

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Cellular agriculture for milk bioactive production

Nature Reviews Bioengineering, Journal Year: 2023, Volume and Issue: 1(11), P. 858 - 874

Published: Oct. 9, 2023

Language: Английский

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