Lacto-N-hexaose (LNH) Production Service
LNH plays an active role in the growth and development of infants, especially in the aspect of shaping the internal microbiota. CD BioGlyco supplies suitable synthesis methods to achieve efficient production of LNH.
Effect of LNH on the Microbial Community
Human milk oligosaccharides (HMOs) are a class of non-conjugated glycans with diverse structures and are the third most abundant solid component of human milk after lipids and lactose. HMO will not be digested by infants but will be used as a prebiotic for beneficial bacteria, thus maintaining the balance of intestinal microflora. In early infancy, the pioneer intestinal microbiota may be susceptible to antibiotics, while the prebiotic oligosaccharides in human milk and some infant formula may protect healthy infant intestinal microbiota. The latest research has proved that two kinds of non-fucosylated HMOs are associated with microbial communities: LNH and 6'-sialyllactose, which are respectively related to the higher and lower relative abundance of Bifidobacterium.
The Exploration Process on LNH
LNH was first isolated and characterized from human milk by the Kobata group in 1927. Later, LNH was also found in other mammals such as Dutch Landrace sows. A recent study found that low concentrations of fucose-L-lactose-N-hexose or LNH were associated with early cessation of breastfeeding. The reports on LNH also include many related derivatives, such as fucosylated LNH. They all have a complex relationship with the development of infant intestinal flora. In the future, it is necessary to conduct further research to determine how LNH affects infant health.
Key Technologies
CD BioGlyco's LNH production services are built upon three core technologies, which can be deployed independently or in combination to meet your project's specific requirements.
- Enzymatic synthesis: We employ a cascade of specific enzymes, such as beta1,4-galactosyltransferase and beta1,3-galactosyltransferase, to sequentially assemble the LNH structure. This approach is particularly advantageous for its high regioselectivity and stereoselectivity, ensuring the production of a structurally pure product.
- Chemoenzymatic synthesis: We combine the precision of chemical synthesis with the efficiency of enzymatic catalysis. The process involves the chemical synthesis of a key tetrasaccharide core, followed by the strategic use of specific enzymes for selective glycosylation.
- Microbial fermentation: By genetically modifying bacterial hosts such as Escherichia coli and yeasts like Saccharomyces cerevisiae, we program them to overexpress key glycosyltransferase genes and utilize inexpensive substrates like lactose to synthesize LNH directly.
LNH Production: Your Partner in Glycan Excellence
CD BioGlyco has accumulated various Synthetic technologies including purification, chemical, enzymatic, chemoenzymatic, and metabolic engineering methods to produce LNH. The optimization strategies we supply include but are not limited to the following:
- Optimized and mature separation and purification technology
- Enzymatic synthesis
- One-pot enzymatic system.
- A pot of multiple enzymes includes bacterial glycosyltransferases and sugar-nucleotide–generating enzymes.
- Selection and identification of suitable enzymes
- Optimal enzyme expression levels.
- Microbial Fermentation
- Construct metabolically engineered strains.
- Optimization of strain culture conditions.
- Chemoenzymatic synthesis
- Combining chemical carbohydrate synthesis with selective enzymatic glycosylation strategy.
- Precursors are obtained by chemical synthesis, then the sequence of galactosyltransferase-catalyzed reactions is rationally arranged.
Workflow
Publication Data
DoI: 10.3389/fchem.2022.905105
Journal: Frontiers in Chemistry
IF: 4.2
Published: 2022
Results: This research presents the initial chemoenzymatic synthesis of LNH. The central approach integrates the chemical synthesis of a crucial branched tetrasaccharide precursor [GlcNH2β1→3(GlcNAcβ1→6)Galβ1→4Glc, featuring a C6-arm GlcNAc and a C3-arm GlcNH2] with selective enzymatic glycosylation steps. Specifically, a β1,4-galactosyl unit was first appended to the C6-arm. Following the conversion of the C3-arm GlcNH2 to GlcNAc through N-acetylation, the Escherichia coli β1,3-galactosyltransferase (WbgO) catalyzed the addition of a β1,4-galactosyl moiety to the C3-arm, thus finalizing the LNH synthesis. Notably, symmetric LNnH was generated as a byproduct during this process. This strategy enables preparative-scale LNH production and could be extended to fucosylated/sialylated branched HMOs for future bioactivity studies.
Fig.1 Enzymatic synthesis of LNH and LNnH. (Ooi, et al., 2022)
Advantages
Deep Expertise in Glycan Engineering
Our team comprises specialists with many years of experience in glycan synthesis and engineering. This deep expertise allows us to navigate the complexities of LNH production and deliver superior results.
Versatile and Custom Synthesis Strategies
By offering a choice of enzymatic, chemoenzymatic, and microbial fermentation methods, we custom-design a solution that perfectly aligns with your project's specific technical and commercial requirements.
Rigorous Quality Control and Purification
Our rigorous, multi-stage purification and quality control protocols ensure that the LNH you receive meets or exceeds the highest industry standards.
Applications
Infant Nutrition
As a functional ingredient in infant formula to mimic the prebiotic effects of human milk.
Pharmaceutical and Therapeutic Development
For research into gut microbiota modulation, immune system regulation, and anti-pathogen strategies.
Functional Foods and Dietary Supplements
As a beneficial prebiotic to promote gut health in children and adults.
Fundamental Research
For studies on glycomics, host-microbe interactions, and the biological functions of HMOs.
Frequently Asked Questions
Customer Review
"The technical support from the CD BioGlyco team on our LNH scale-up was exceptional. They provided a consistent, ultra-high-purity batch that immediately passed our stringent quality control checks. Their detailed analytical reports provided total confidence in the product's asymmetric structure."
- J.F., Nutraceutical Development Lab
"The transition from our R&D-scale LNH synthesis to a commercial quantity was seamless thanks to CD BioGlyco's robust enzymatic process. The yield and cost-efficiency achieved were significantly better than our previous attempts with pure chemical methods."
- S.K., Food Science Institute
"We required LNH with a specific linker for a high-throughput diagnostic assay. CD BioGlyco delivered the custom-derivatized product with confirmed linkage and over 99% purity. Their speed and precision are truly best-in-class, making them our preferred supplier for complex glycan standards."
- A.R., Diagnostic Technology Firm
Associated Services
CD BioGlyco has made many achievements in the field of production services of LNH, which are widely praised by clients internationally. If you have any further queries, please contact us directly for more details.
Reference
- Ooi, K.E.; et al. Chemoenzymatic synthesis of asymmetrically branched human milk oligosaccharide lacto-N-hexaose. Frontiers in Chemistry. 2022, 10: 905105. (Open Access)
