Regulation of Metabolic Pathways to Enhance Difucosyllactose Biosynthesis in Escherichia coli DOI

Shanquan Liang,

Qi Quan,

Dan Liu

et al.

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

Published: Dec. 19, 2024

Difucosyllactose (DFL), an important kind of fucosylated human milk oligosaccharides (HMOs), has garnered considerable attention due to its excellent physiological activities in infants. Previously, we achieved de novo biosynthesis DFL; however, substantial residual intermediates fucosyllactoses (FL) were detected. In this study, DFL was optimized, and FL reduced by regulating metabolic pathways. Different plasmid combinations used regulate gene expression, achieving optimal flux balance between 2′-FL DFL. The expression level key enzyme α-1,3-fucosyltransferase (α-1,3-FT, FucTa) then enhanced increasing copy number integrating fucTa into the chromosome. Exocytosis deleting sugar efflux transporter setA gene, thereby minimizing FL. Finally, strain BSF41 produced 55.3 g/L with only 2.59 a 5 L fermentor, representing highest reported titer date. This study provides foundation for advancing HMOs.

Language: Английский

Recent advances in the science of human milk oligosaccharides DOI Creative Commons
Tadasu Urashima, Katsumi Ajisaka, Tetsuro Ujihara

et al.

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

Published: Jan. 1, 2025

Language: Английский

Citations

1

Synthesising human milk oligosaccharide using biotechnology-driven enzymes DOI

Jayashree Niharika,

Ravindra Pal Singh

Systems Microbiology and Biomanufacturing, Journal Year: 2025, Volume and Issue: unknown

Published: Jan. 28, 2025

Language: Английский

Citations

1

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

Mengting Tao,

Long-Hao Yang, Chunhua Zhao

et al.

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

Citations

7

High-Level Production of l-Fucose by Plasmid-Free or Antibiotic-Independent Metabolically Engineered Escherichia coli Strains DOI
Jiawei Meng, Yingying Zhu, Zhen Lu

et al.

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

Citations

4

Efficient production of 3-fucosyllactose in a plasmid-free engineered Escherichia coli through strengthening of the GDP-fucose pathway DOI
Yihan Chen, Yingying Zhu, Hao Wang

et al.

Food Bioscience, Journal Year: 2024, Volume and Issue: 59, P. 104154 - 104154

Published: April 19, 2024

Language: Английский

Citations

4

Multidimensional Engineering of Escherichia coli MG1655 for the Efficient Biosynthesis of Difucosyllactose DOI
Yihan Chen, Chunhua Zhao,

Ruiyan Wang

et al.

Journal of Agricultural and Food Chemistry, Journal Year: 2025, Volume and Issue: 73(9), P. 5405 - 5413

Published: Feb. 24, 2025

Difucosyllactose (DFL), a representative bisfucosylated oligosaccharide found in human milk, has garnered significant attention due to its immense health benefits. To date, several plasmid-based engineered strains have been established for DFL synthesis. However, these face challenges such as antibiotic dependence and plasmid instability, which limit their commercial application the food industry. For this, plasmid-free Escherichia coli MG1655 were by integrating multicopy numbers of SAMT fut3Bc into genome overexpressing key genes involved GDP-l-fucose pathway. enhance catalytic efficiency, Fut3Bc was mutated based on AlphaFold 3, obtaining beneficial mutant (F24Y). The optimized strain, MGA2S-5, containing 2 copies 4 (F24Y) obtained. Eventually, strain MGA2S-5 synthesized 35.04 g/L 7 L bioreactor fed-batch cultivation, with no intermediate products remaining medium.

Language: Английский

Citations

0

Glycosyltransferases in human milk oligosaccharide synthesis: structural mechanisms and rational design DOI
Yanfeng Liu, Guocheng Du, Jian Chen

et al.

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

Published: May 9, 2025

Language: Английский

Citations

0

Serratia marcescens ATCC 274 increases production of the red pigment prodigiosin in response to Chi phage infection DOI Creative Commons
Nathaniel C. Esteves, Birgit E. Scharf

Scientific Reports, Journal Year: 2024, Volume and Issue: 14(1)

Published: July 31, 2024

Serratia marcescens is an opportunistic human pathogen that produces a vibrant red pigment called prodigiosin. Prodigiosin has implications in virulence of S. and promising clinical applications. We discovered addition the virulent flagellotropic bacteriophage χ (Chi) to culture stimulates greater than fivefold overproduction Active phage infection required for effect, as χ-resistant strain lacking flagella does not respond presence. Via reporter fusion assay, we have determined χ-induced cell lysate uninfected causes threefold increase transcription pig operon, containing genes essential biosynthesis. Replacement promoter with constitutive abolished pigmentation increase, indicating regulatory elements present likely mediate phenomenon. hypothesize detects threat phage-mediated death reacts by producing prodigiosin stress response. Our findings are significance two main reasons: (i) elucidating complex phage-host interactions crucial development therapeutic treatments, (ii) response could be exploited its biosynthesis use pharmaceutical.

Language: Английский

Citations

3

Multistrategy Optimization for High-Yield 3-Fucosyllactose Production in Escherichia coli BL21 Star (DE3) DOI

LU Shu-jie,

Caiwen Lao,

Jin Wang

et al.

Journal of Agricultural and Food Chemistry, Journal Year: 2025, Volume and Issue: 73(9), P. 5385 - 5394

Published: Feb. 19, 2025

3-Fucosyllactose (3-FL), an essential component of human milk oligosaccharides, is crucial for infant health. However, the extraction 3-FL from presents a significant challenge. In this study, E. coli BL21 Star (DE3) was engineered biosynthesis, and gene combinations metabolic pathways were optimized to obtain high-yield production strain. First, module integrated into coli. Second, efficient alpha-1,3-fucosyltransferase screened. Then, different integration sites used optimize precursor synthesis cluster transferase genes screen transporter proteins glycerol utilization that could enhance production. The 3FL05-1 strain yielded titer 11.26 g/L in shake flasks. Further amplification 5 L bioreactor resulted 60.24 g/L, which represents highest reported date.

Language: Английский

Citations

0

Developing a Metabolic Flux Optimized Escherichia coli for Efficient Production of Difucosyllactose DOI
Yihan Chen, Yingying Zhu, Hao Wang

et al.

ACS Sustainable Chemistry & Engineering, Journal Year: 2024, Volume and Issue: 12(38), P. 14249 - 14257

Published: Sept. 9, 2024

Language: Английский

Citations

2