De Novo Biosynthesis of Difucosyllactose by Artificial Pathway Construction and α1,3/4-Fucosyltransferase Rational Design in .

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 of GDP-fucose from only glycerol. This was achieved by engineering to endogenously overexpress genes , , , and and heterologously overexpress a pair of fucosyltransferases to produce DFL from lactose.

Engineering MG1655 for Highly Efficient Biosynthesis of 2'-Fucosyllactose by GDP-Fucose Pathway.

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 a GDP-fucose pathway from engineered MG1655. First, and , two competitive pathway genes, were disrupted to block the invalid consumption of lactose and GDP-fucose, respectively.

Elimination of Byproduct Generation and Enhancement of 2'-Fucosyllactose Synthesis by Expressing a Novel α1,2-Fucosyltransferase in Engineered .

2'-Fucosyllactose (2'-FL) is a kind of fucosylated human milk oligosaccharide (HMO), representing the most abundant oligosaccharide in breast milk. We conducted systematic studies on three canonical α1,2-fucosyltransferases (WbgL, FucT2, and WcfB) to quantify the byproducts in a - and -deleted BL21(DE3) basic host strain. Further, we screened a highly active α1,2-fucosyltransferase from sp.

De novo biosynthesis of 2'-fucosyllactose in a metabolically engineered Escherichia coli using a novel ɑ1,2-fucosyltransferase from Azospirillum lipoferum.

Human milk oligosaccharides are complex, indigestible oligosaccharides that provide ideal nutrition for infant development. Here, 2'-fucosyllactose was efficiently produced in Escherichia coli by using a biosynthetic pathway. For this, both lacZ and wcaJ (encoding β-galactosidase and UDP-glucose lipid carrier transferase, respectively) were deleted to enhance the 2'-fucosyllactose biosynthesis.

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

l-Fucose is a natural deoxy hexose found in a variety of organisms. It possesses many physiological effects and has potential applications in pharmaceutical, cosmetic, and food industries. Microbial synthesis via metabolic engineering attracts increasing attention for efficient production of important chemicals.

Spatial organization of pathway enzymes via self-assembly to improve 2'-fucosyllactose biosynthesis in engineered Escherichia coli.

As one of the most abundant components in human milk oligosaccharides, 2'-fucosyllactose (2'-FL) possesses versatile beneficial health effects. Although most studies focused on overexpressing or fine-tuning the expression of pathway enzymes and achieved a striking increase of 2'-FL production, directly facilitating the metabolic flux toward the key intermediate GDP-l-fucose seems to be ignored. Here, multienzyme complexes consisting of sequential pathway enzymes were constructed by using specific peptide interaction motifs in recombinant Escherichia coli to achieve a higher titer of 2'-FL.

Interaction between the Anchoring Domain of A-Kinase Anchoring Proteins and the Dimerization and Docking Domain of Protein Kinase A: A Potent Tool for Synthetic Biology.

Nature is enriched with specific interactions between receptor proteins and their cognate ligands. These interacting pairs can be exploited and applied for the construction of well-ordered multicomponent assemblies with multivalency and multifunctionality. One of the research hotspots of this area is the formation of multienzyme complexes with stable and tunable architectures, which may bear the potential to facilitate cascade biocatalysis and/or strengthen metabolic fluxes.

High-Level Biosynthesis of 2'-Fucosyllactose by Metabolically Engineered .

2'-Fucosyllactose (2'-FL) is the most abundant oligosaccharide in human milk. In this study, a highly efficient biosynthetic route for 2'-FL production was designed the pathway of GDP-l-fucose using engineered BL21(DE3). Specifically, plasmid-based strains with previously deleted and were further reconstructed by introducing pathway genes and α1,2-fucosyltransferase-encoding to realize 2'-FL synthesis.