CD BioGlyco has various technologies to analyze HMOs. We work with advanced technology along with dedicated professionals to provide you with high-quality products and services.
HMOs are a class of over 200 compounds present at 20-23 g/L in colostrum and 12-14 g/L in mature milk. Unlike their common precursor Lactose, HMOs are indigestible by human infants and instead improve neonatal health by serving as effective Antimicrobials and Antivirals, prebiotics, and regulators of inflammatory immune cell-response cascades. These and other potential benefits of HMOs make them attractive targets of study for preventing or treating diseases in both children and adults. The bioactive properties of HMOs have motivated efforts to define the mechanistic effects of individual compounds.
DFL is a fucosyloligosaccharide present in human milk and colostrum. DFL is one of the most abundant fucosylated HMOs and is produced at an average of 0.43 g/L over the first year of lactation by secretory mothers. DFL, a tetrasaccharide, constitutes d-glucose, d-galactose, and L-fucose. Its structure consists of a core lactose unit that is fucosylated at the C2 ' and C3 positions.
Fig.1 The structure of DFL. (CD BioGlyco)
DFL has strong antimicrobial activity against various pathogens, including group B Streptococcus, identified as the leading cause of neonatal sepsis. Of the purified HMOs tested, only DFL significantly inhibits the thrombin-induced release of the pro-inflammatory proteins RANTES and sCD40L. Besides, studies have shown that DFL is effective in preventing Campylobacter jejuni associated diarrhea in neonates. Its activities as a gastrointestinal and immunological modulator have motivated further research into its potential therapeutic applications.
Microbial production is a viable method to produce DFL. Whole-cell biocatalysts are self-maintaining systems and do not require an exogenous supply of expensive cofactors. Enzymatic reactions in cells can also achieve high regio- and stereo-specific production of structurally complex molecules. A system is created in E. coli using two fucosyltransferases that preferentially fucosylated lactose to form a 2 '-FL intermediate that is further fucosylated to produce the target DFL. The strain produces 5.1 g/L of DFL from 3 g/L lactose, achieving 91% of the theoretical maximum yield of DFL in 24 h.
We analyze DFL by capillary electrophoresis (CE), which is one of the most effective glycoanalytical ways widely used in the pharmaceutical, biomedical, and food industries. The sensitivity of CE-based analysis with laser-induced fluorescent detection (LIF) can be as low as 10-10 M after tagging the oligosaccharides with a fluorescent label such as aminopyrenetrisulfonate and APTS. APTS also offers charge to the otherwise mostly neutral carbohydrates to operate electromigration. Besides, all reducing sugars are labeled at their reductive end only. The one APTS/sugar structure makes it easy quantification. The secondary equilibrium of the borate-diol complexation may further improve resolving power with the use of borate-containing buffer systems. Capitalizing on carbohydrates and linkage-specific exoglycosidase-based digestions, the sequence of the oligosaccharide structures is determined.
CD BioGlyco is a professional company in the field of glycobiology that is committed to helping clients to solve problems. Please feel free to contact us directly if you need technical support, and we will try our best to provide you with the best solution.
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