Enzyme and Microbial Technology, Journal Year: 2024, Volume and Issue: 184, P. 110576 - 110576
Published: Dec. 25, 2024
Language: Английский
Systems Microbiology and Biomanufacturing, Journal Year: 2025, Volume and Issue: unknown
Published: Jan. 28, 2025
Language: Английский
Citations
1
Journal of Agricultural and Food Chemistry, Journal Year: 2024, Volume and Issue: 72(19), P. 11013 - 11028
Published: May 1, 2024
Five GH29B α-1,3/4-l-fucosidases (EC 3.2.1.111) were investigated for their ability to catalyze the formation of human milk oligosaccharide lacto-N-fucopentaose II (LNFP II) from lacto-N-tetraose (LNT) and 3-fucosyllactose (3FL) via transglycosylation. We studied effect pH on transfucosylation hydrolysis explored impact specific mutations using molecular dynamics simulations. LNFP yields 91 65% obtained wild-type SpGH29C CpAfc2 enzymes, respectively, being highest transglycosylation reported date. BbAfcB BiAfcB are highly hydrolytic enzymes. The results indicate that effects buffer systems enzyme-dependent yet relevant consider when designing reactions. Replacing Thr284 in with Val resulted increased yields, while opposite replacement Val258 Val289 Thr decreased transfucosylation, confirming a role controlling flexibility acid/base loop which turn affects substitution an Ala residue His almost abolished secondary BbAfcB. directly applicable enhancement may have significant implications manufacturing as new infant formula ingredient.
Language: Английский
Citations
8
Journal of Agricultural and Food Chemistry, Journal Year: 2024, Volume and Issue: unknown
Published: April 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
Language: Английский
Citations
5
Journal of Agricultural and Food Chemistry, Journal Year: 2024, Volume and Issue: unknown
Published: Dec. 10, 2024
Nicotinamide mononucleotide (NMN), a key precursor of NAD
Language: Английский
Citations
5
Journal of Agricultural and Food Chemistry, Journal Year: 2024, Volume and Issue: 72(18), P. 10469 - 10476
Published: April 25, 2024
Lacto-N-difucohexaose II (LNDFH II) is a typical fucosylated human milk oligosaccharide and can be enzymatically produced from lacto-N-tetraose (LNT) by specific α1,3/4-fucosyltransferase Helicobacter pylori DMS 6709, referred to as FucT14. Previously, we constructed an engineered Escherichia coli BL21(DE3) with single plasmid for highly efficient biosynthesis of LNT. In this study, two additional plasmids harboring the de novo GDP-L-fucose pathway module FucT14, respectively, were further introduced construct strain successful LNDFH II. FucT14 was actively expressed, major product, lacto-N-fucopentaose (LNFP) V minor trace amount LNFP 3-fucosyllactose very products. Additional expression α1,3-fucosyltransferase FutM1 Bacteroidaceae bacterium gut metagenome could obviously enhance biosynthesis. After optimization induction conditions, maximum titer reached 3.011 g/L shake-flask cultivation. During fed-batch cultivation, efficiently highest 18.062 productivity yield 0.301 g/L·h.
Language: Английский
Citations
4
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
Citations
4
Journal of Agricultural and Food Chemistry, Journal Year: 2025, Volume and Issue: 73(9), P. 5376 - 5384
Published: Feb. 18, 2025
Biological production of human milk oligosaccharides (HMOs) using metabolically engineered strains is a research hotspot in food biotechnology, but less effort has been made on the biological sialylated complex HMOs. Sialyllacto-N-tetraose c only monosialylated HMO top 15 In this study, metabolic pathway LST was constructed Escherichia coli BL21(DE3) by introducing three sequential glycosyltransferases: β1,3-N-acetylglucosaminyltransferase, β1,4-galactosyltransferase, and α2,6-sialyltransferase. The cytidine 5′-monophospho (CMP)-N-acetylneuraminic acid (Neu5Ac) enhanced to improve production. β1,4-galactosyltransferase from Helicobacter pylori J99 (HpGalT) α2,6-sialyltransferase Vespertiliibacter pulmonis (ED6ST) were screened as pair key glycosyltransferases for enhancing final strain could produce 1.718 9.745 g/L shake-flask fed-batch cultivation, respectively, indicating feasibility efficient biosynthesis
Language: Английский
Citations
0
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
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
Journal of Agricultural and Food Chemistry, Journal Year: 2025, Volume and Issue: unknown
Published: March 4, 2025
Recently, the construction of metabolically engineered strains for microbial synthesis human milk oligosaccharides (HMOs) has attracted increasing attention. However, fewer efforts were made in vivo biosynthesis complex HMOs, especially sialylated HMOs. In this study, we Escherichia coli BL21(DE3) to efficiently produce sialyllacto-N-tetraose a (LST-a) efficiently. Three sequential glycosylation steps introduced construct LST-a pathway, catalyzed by β1,3-N-acetylglucosaminylation, β1,3-galactosylation, and α2,3-sialylation. Pathway genes cytidine 5'-monophospho (CMP)-N-acetylneuraminic acid (Neu5Ac) support sialylation donor supply. Production was improved deleting competitive CMP-Neu5Ac synthesis, screening more efficient α2,3-sialyltransferase, combinatorial optimization pathway gene expression. finally produced with titer 1.235 4.85 g/L shake-flask fed-batch cultivation, respectively, demonstrating feasibility production
Language: Английский
Citations
0