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

---

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

---

Efficient Biosynthesis of Difucosyllactose via De Novo GDP-l-Fucose Pathway in Metabolically Engineered Escherichia coli

Journal of Agricultural and Food Chemistry, Journal Year: 2024, Volume and Issue: 72(8), P. 4367 - 4375

Published: Feb. 20, 2024

Difucosyllactose (DFL) is an important component of human milk oligosaccharides (HMOs) and has significant benefits for the growth development infants. So far, a few microbial cell factories have been constructed production DFL, which still problems low high cost. Herein, high-level de novo pathway DFL-producing strain was by multistep optimization strategies in Escherichia coli BL21star(DE3). We first efficiently synthesized intermediate 2′-fucosyllactose (2′-FL) E. BL21star(DE3) advisable stepwise strategy. The truncated α-1,3/4-fucosyltransferase (Hp3/4FT) then introduced into engineered to achieve biosynthesis DFL. ATP-dependent protease (Lon) GDP-mannose hydrolase (NudK) were deleted, mannose-6-phosphate isomerase (ManA) overexpressed improve GDP-l-fucose accumulation. regulator RcsA fine-tune expression level genes, thereby increasing synthesis final produced 6.19 g/L DFL shake flask 33.45 5 L fermenter, highest reported titers so far. This study provides more economical, sustainable, effective strategy produce fucosylated (HMOs).

Language: Английский

---

Implementation of a Quorum-Sensing System for Highly Efficient Biosynthesis of Lacto-N-neotetraose in Engineered Escherichia coli MG1655

Journal of Agricultural and Food Chemistry, Journal Year: 2024, Volume and Issue: 72(13), P. 7179 - 7186

Published: March 23, 2024

Lacto-N-neotetraose (LNnT), a prominent neutral human milk oligosaccharide (HMO), serves as pivotal structural element in complex HMO biosynthesis. Given its promising health effects for infants, the biosynthesis of LNnT is garnering greater interest. Using previously engineered strain chassis, highly effective producer was constructed. First, synthesis Escherichia coli MG1655 achieved by introducing β1,3-N-acetylglucosaminyltransferase LgtA and β1,4-galactosyltransferase CpsIaJ, coupled with optimization enzyme expression levels using various promoters. Subsequently, ugd underwent disruption, galE gene enhanced replacing promoter PJ23119 or Ptac. Then, lux-type quorum sensing (QS) system applied to achieve varied metabolic regulation. Additionally, systematic QS promoters conducted further improve titer shake flask. Finally, extracellular 20.33 g/L, accompanied productivity 0.41 g/L/h.

Language: Английский

---

Recent advances in the science of human milk oligosaccharides

BBA Advances, Journal Year: 2025, Volume and Issue: unknown, P. 100136 - 100136

Published: Jan. 1, 2025

Language: Английский

---

Engineering Pichia pastoris for Efficient De Novo Synthesis of 2′-Fucosyllactose

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

Published: March 28, 2025

2′-Fucosyllactose (2′-FL), the most abundant in human milk oligosaccharides (HMOs), is a nutrient of great importance. As safe organism widely used industries, Pichia pastoris was tested here for 2′-FL production. The de novo biosynthesis pathway constructed using genome-editing technology based on CRISPR-Cas9 with an initial titer 1.01 g/L. Introducing N-terminal SUMO or Ub tag to FucT2 and transporter CDT2 from Neurospora crassa into P. found improve Then, modular metabolic engineering conducted production, enhancing GTP supply module, NADPH regeneration precursor module. Subsequently, key enzyme semirationally designed further increase Finally, production by engineered scaled up 3 L fermenter fed-batch mode, resulting 22.35 g/L that highest pastoris. results prove effectiveness strategies demonstrate could be potential chassis produce HMOs.

Language: Английский

---

High-Yield Synthesis of 2′-Fucosyllactose from Glycerol and Glucose in Engineered Escherichia coli

Journal of Agricultural and Food Chemistry, Journal Year: 2023, Volume and Issue: 71(41), P. 15237 - 15248

Published: Oct. 5, 2023

2'-Fucosyllactose (2'-FL) is vital for the growth and development of newborns. In this study, we developed a synthesis pathway 2'-FL in Escherichia coli BL21 (DE3). Then, optimized solubility α-1,2-fucosyltransferase, thereby enhancing production yield 2'-FL. Based on finding, further enhanced expression guanosine inosine kinase Gsk knocked out isocitrate lyase regulator gene iclR. This strategy reduced formation byproduct acetate during metabolic process alleviated carbon source overflow effects strain, resulting improvement 3 L bioreactor, employing fed-batch fermentation with glycerol glucose as substrates, engineered strain BWLAI-RSZL exhibited impressive titers 121.9 111.56 g/L, along productivity levels 1.57 1.31 g/L/h, respectively. The reported reached groundbreaking level, irrespective employed (glycerol or glucose), highlighting significant potential large-scale industrial

Language: Английский

---

Synthesis of bioactive oligosaccharides and their potential health benefits

Critical Reviews in Food Science and Nutrition, Journal Year: 2023, Volume and Issue: 64(28), P. 10319 - 10331

Published: June 21, 2023

Oligosaccharides, a low polymerization degree of carbohydrate, possess various physiological activities, such as anti-diabetes, anti-obesity, anti-aging, anti-viral, and gut microbiota regulation, having widely used in food medical fields. However, due to the limited natural oligosaccharides, many un-natural oligosaccharides from complex polysaccharides are being studied for amplifying available pool oligosaccharides. More recently, were developed by using several artificial strategies, chemical degradation, enzyme catalysis, biosynthesis, then they can be applied sectors. Moreover, it has gradually become trend use biosynthesis realize synthesis with clear structure. Emerging research found that exert more comprehensive effects against human diseases through multiple mechanisms. these routes have not been critical reviewed summarized. Therefore, purpose this review is present preparations healthy effects, focus on diabetes, obesity, aging, virus, microbiota. Additionally, application multi-omics also discussed. Especially, needed apply disease models find out biomarkers respond dynamic change process

Language: Английский

---

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

---

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

---