Síntesis de fucooligosacáridos empleando α-L-fucosidasas microbianas: estudio in vitro e in silico DOI Creative Commons

Mauricio Eduardo Pavón Chimal

Published: Dec. 13, 2023

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
Optimization of l-Fucose Biosynthesis in Escherichia coli through Pathway Engineering and Mixed Carbon Source Strategy DOI

Zhu Xia,

Caiwen Lao,

Jinyong Wu

et al.

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

Published: March 3, 2025

This study presents an engineered strain of Escherichia coli specifically designed to enhance the production l-fucose while minimizing residues 2′-fucosyllactose. The optimization strategies employed include selection key enzymes, gene copy numbers, and fermentation using mixed carbon sources. metabolic flux was directed toward synthesis by integrating preferred 1,2-fucosyltransferase α-l-fucosidase into genome. Furthermore, numbers were optimized enzyme expression, thereby increasing production. Additionally, supply guanosine 5′-triphosphate improved, cofactors regenerated better regulate metabolism. Modifications transporter proteins effectively reduced accumulation implementation a glucose/glycerol co-fermentation strategy enhanced distribution efficiency. achieved yield 91.90 g/L in 5 L bioreactor, representing 80.01% increase over previous yields, with productivity 1.18 g L–1 h–1. is highest reported for l-fucose, demonstrating its potential industrial

Language: Английский

Citations

0
Dual Antibiotic-Free Plasmid Systems Enable High-Efficiency l-Fucose Biosynthesis DOI
Jiawei Meng, Yingying Zhu, Zhen Lu

et al.

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

Citations

0
Recent advances in a functional deoxy hexose l-fucose: Occurrence, physiological effects, and preparation DOI
Jiawei Meng, Yingying Zhu, Ningning Wang

et al.

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

Published: May 29, 2023

Language: Английский

Citations

9
Efficient Biosynthesis of Lacto-N-Biose I, a Building Block of Type I Human Milk Oligosaccharides, by a Metabolically Engineered Escherichia coli DOI

Mengting Tao,

Long-Hao Yang, Chunhua Zhao

et al.

Journal of Agricultural and Food Chemistry, Journal Year: 2024, Volume and Issue: 72(11), P. 5860 - 5866

Published: March 7, 2024

Lacto-N-biose I (LNB), termed a Type 1 disaccharide, is an important building block of human milk oligosaccharides. It shows promising prebiotic activity by stimulating the proliferation many gut-associated bifidobacteria and thus displays good potential in infant foods or supplements. Enzymatic microbial approaches to LNB synthesis have been studied, almost all which involve glycosylation phosphorylase as final step. Herein, we report new easier strategy through route "lactose → lacto-N-triose II (LNTri II) lacto-N-tetraose (LNT) LNB". A previously constructed LNT-producing Escherichia coli BL21(DE3) strain was engineered for biosynthesis introducing Bifidobacterium bifidum LnbB. efficiently produced, accompanied lactose regeneration. Genomic integration key pathway genes related LNTri LNT performed enhance titers. The produced 3.54 26.88 g/L shake-flask fed-batch cultivation, respectively.

Language: Английский

Citations

2
Research progress on the functions, preparation and detection methods of l-fucose DOI
Yan Wang, Xiao Zhang, Xiao Tian

et al.

Food Chemistry, Journal Year: 2023, Volume and Issue: 433, P. 137393 - 137393

Published: Sept. 4, 2023

Language: Английский

Citations

5
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

Citations

1
Recent Progress in Physiological Significance and Biosynthesis of Lacto-N-triose II: Insights into a Crucial Biomolecule DOI

Cuie Guang,

Zhihui Du, Jiawei Meng

et al.

Journal of Agricultural and Food Chemistry, Journal Year: 2024, Volume and Issue: 72(36), P. 19539 - 19548

Published: Aug. 27, 2024

Lacto-

Language: Английский

Citations

0
Virus-bacterium interaction involved in element cycles in biological treatment of coking wastewater DOI
Zhi-Jie Tan,

Wanling Chen,

Xinyi Wei

et al.

Bioresource Technology, Journal Year: 2024, Volume and Issue: unknown, P. 131839 - 131839

Published: Nov. 1, 2024

Language: Английский

Citations

0
Síntesis de fucooligosacáridos empleando α-L-fucosidasas microbianas: estudio in vitro e in silico DOI Creative Commons

Mauricio Eduardo Pavón Chimal

Published: Dec. 13, 2023

Citations

0