The oligosaccharide fraction is the third largest component of human milk after lactose and lipids. At CD BioGlyco, we pay attention to every detail in HMO Production and strive for perfection in every process, evidenced by our many years of successful experience.

Overview of Human Milk Oligosaccharides (HMO)

Most milk carbohydrates have a lactose (Lac) unit at the reducing end. In addition to Lac, several core structures have been discovered so far, such as lacto-N-tetraose (LNT), lacto-N-neotetraose (LNnT), lacto-N-hexaose (LNH), three further hexaoses, several octaoses, and even decaoses. These core structures are built from Lac by adding lactone-biose sugar units, Galβ1-3GlcNAc (type 1 structure), or lactosamine units, Galβ1-4GlcNAc (type 2 structure). Generally, these disaccharide units are either β1,3 linked to each other or Lac to form elongated linear structures, or β1,6 linked to internal galactose residues to form branched oligosaccharides. These core structures are substrates for various fucosyltransferases (FucT) and sialyltransferases in mammary epithelial cells. Various focused carbohydrates are biosynthesized in various epithelial cells due to specific actions of several human α-FucTs.

Chemical Structure of Fucosyl-lacto-N-neohexaose I (FLNnH I)

FLNnH I is a human milk oligosaccharide. The chemical structure is shown in Fig.1.

Fig.1 Chemical structure of FLNnH I. (CD BioGlyco)

FLNnH I Production Service at CD BioGlyco

CD BioGlyco provides an enzymatic synthetic method to produce FLNnH I, which is achieved using a continuous one-pot enzymatic system. This method involves bacterial glycosyltransferases and corresponding sugar-nucleotide–generating enzymes to facilitate the efficient production of N-acetyllactosamine (LacNAc) backbone and heterochains. The fucosyltransferase used is critical for the efficiency of fucosylation with respect to the actual synthesis of fucosylated glycans. The process includes the following steps:

• The 6-azidohexyllactidine acceptor is first prepared, and the azide group facilitates the immobilization of the compound on the glass slide, thereby further developing the glycan array.
• The lacto-N-tetraose is synthesized by WbgO-catalyzed sequential one-pot galactosylation and in situ UDP-Gal preparation.
• Using a similar approach to the sequential one-pot synthesis, HMOs containing N-LacNAc repeats are synthesized.

For the technology of enzymatic synthesis of oligosaccharides, please refer to Production of HMO by Chemoenzymatic Methods.

In addition, CD BioGlyco offers a range of methods for HMO Profiling. We provide high-performance liquid chromatography (HPLC) for the measurement of FLNnH I in milk samples (with appropriate pretreatment). Retention times are compared to those of commercial standard oligosaccharides and quantified based on response factors. In addition, we calculate the relative abundance of HMOs.

Advantages of Us

• A continuous one-pot enzymatic system for the enzymatic synthesis of FLNnH I
• Based on retention time comparison with commercial standard FLNnH I and MS analysis
• Outstanding scientists specialize in HMO production
• Detailed and reliable after-sales service

CD BioGlyco has advanced instrumentation platforms and expertise in glycoscience. In addition, our professional research teams allow us to provide clients with high-quality services for FLNnH I production and HMO profiling. If you are interested in our services, please contact us for more details without any hesitation.

Reference:

1. Moossavi, S.; et al. Integrated analysis of human milk microbiota with oligosaccharides and fatty acids in the CHILD cohort. Frontiers in nutrition. 2019, 6: 58.