Lacto-N-tetraose (LNT) Production Service
Through competent assistance by our researchers and the fastest delivery of high-quality lacto-N-tetraose (LNT) products at fair prices, CD BioGlyco has become a "preferred supplier" of many clients. We have developed many effective strategies to deal with the uncertainty of metabolic patterns in model microorganisms.
Relationship Between Human Milk and LNT
Human milk oligosaccharides (HMOs), the third largest molecular constituent in breast milk, have received considerable attention for their importance to infant health, much of which stems from the physiological factor that they can promote the development of a healthy microbiome. LNT is a core structure shared by long-chain HMOs and many glycolipids, its content is 8 times that of Lacto-N-neotetraose.
Safety of LNT in Food Applications
More accurately mimicking the composition of human breast milk is certainly a production priority in commercial infant formula, which requires the addition of naturally occurring structural products identical to the milk oligosaccharides produced by humans. One study specifically evaluated the safety of LNT, as part of an in vitro chronic subchronic gavage toxicity study and genotoxicity testing. Study data showed no compound-related adverse effects. This supports the safe use of LNT in infant formula and the corresponding composition ratio.
Key Technologies
A promising method is enzymatic synthesis, especially via glycosyltransferases, which provides a highly specific and efficient route. This approach relies on enzymes to accurately construct glycan frameworks, showcasing remarkable stereo- and regioselectivity that traditional methods struggle to match. For LNT biosynthesis, this strategy commonly integrates advanced genetic engineering to engineer key enzymes, followed by microbial fermentation for scalable production. Notably, this method enables the generation of LNT with remarkable purity and yield, highlighting its technical superiority in glycan synthesis.
Your Partner for LNT Production Innovation
Due to the development of metabolic engineering and synthetic biology strategies, the microbial production of LNT provided by us has developed rapidly. Our researchers have accumulated mature production technologies and are constantly improving and innovating, including but not limited to the following:
- Synthesis by chemoenzymatic methods
- First synthesizing lacto-N-triose by chemical method. Then, using it as an acceptor, a molecule of galactose is introduced into the lacto-N-triose acceptor through a one-pot two-enzyme reaction to synthesize LNT.
- Lactate-N-disaccharide derivative is prepared by the chemoenzymatic method as donor substrate, and LNT was synthesized by glycosylation of wild-type and glycosidase variants from Bifidobacterium bifidum.
- Biological production methods
- Construct engineered Escherichia coli.
The transferases of exogenous strains and derived Escherichia coli are simultaneously introduced into other derived Escherichia coli strains to construct engineered Escherichia coli. - The pathway is optimized through genome combination, fine-tuning, overexpression, and other strategies.
- Scale up production using shake flask fermentation and fed-batch.
- Construct engineered Escherichia coli.
Biological Strategies to Optimize Production at CD BioGlyco
- Improve intracellular lactose supply
Modifying lactose permease through rational design and directed evolution, overexpressing permease to improve intracellular lactose supply.
- Genetic engineering
Deletion of specific coding genes enables simultaneous consumption of glucose and lactose. Screening genes related to LNT metabolic pathway and inactivating genes related to the bypass pathway.
- Adjust dynamic balance
Inhibiting or increasing the expression of genes necessary for homologous or heterologous pathway balance in strains such as Escherichia coli and Bacillus subtilis to reduce the impact on cell growth, etc.
Workflow
Publication Data
Journal: Frontiers in nutrition
IF: 5.1
Published: 2018
Results: The article explores how the gut bacterium Bifidobacterium longum subsp. infantis metabolizes two specific HMO isomers, LNT and lacto-N-neotetraose (LNnT). The study found that while both LNT and LNnT are utilized, the less efficient metabolism of LNnT forces the bacterium to shift its carbon metabolism, resulting in the secretion of different byproducts like formic acid and ethanol. This metabolic variation, stemming from a single glycosidic linkage difference between the two HMOs, highlights how the structure of milk glycans can influence microbial physiology and potentially impact infant gut health.
Fig.1 Bifidobacterium longum subsp. infantis metabolic pathways for utilization of LNT and LNnT and their constituent monosaccharides. (Özcan & Sela, 2018)
Advantages
Exceptional Purity via Optimized Synthesis
Our optimized enzymatic synthesis and advanced purification processes consistently deliver LNT with exceptional purity.
Flexible and Seamless Scalability
We offer a production platform that scales seamlessly from milligram-scale research quantities to large-scale industrial volumes, with the flexibility to adapt to your project's unique requirements.
Stringent Quality Control and Compliance
Every step of our workflow is governed by stringent quality control protocols to ensure product consistency and regulatory compliance.
Applications
Dietary Supplements for Immune and Digestive Support
Incorporated into dietary supplements, LNT supports a balanced gut microbiota, which is linked to a host of health benefits from improved digestion to enhanced immune function.
Functional Food and Beverage Development
LNT can be used as a prebiotic ingredient in a variety of food and beverage development, including yogurts, juices, and health drinks, to enhance gut health and digestive wellness.
Infant Formula Fortification
As a cornerstone of HMOs, LNT is crucial for fortifying infant formulas to more closely mimic the composition and benefits of human breast milk, promoting healthy gut maturation and immune system development.
Frequently Asked Questions
Customer Review
"The LNT we received was essential for our study. The purity levels exceeded the CoA specification, which was critical for ensuring the accurate interpretation of our gut microbiome data. The stability and documentation provided made the paperwork preparation straightforward and efficient. We are very satisfied with CD BioGlyco's technical support."
- Dr. X.C., R&D Director
"Our challenge was scaling up production without introducing cost-prohibitive purification steps. CD BioGlyco's engineered biosynthesis approach solved this. Their batch consistency was remarkable across multiple kilogram-scale orders, proving their robust manufacturing control."
- J.R., Head of Operations
"We required a custom quantity of LNT for a long-term academic study into host-pathogen interactions. The team at CD BioGlyco was flexible, responsive, and provided timely delivery. Their deep understanding of the glycan chemistry ensured that the product was delivered exactly to our required analytical specification, enabling us to advance our research findings."
- Prof. A.S., Principal Investigator
Associated Services
CD BioGlyco uses the mature experience and technology accumulated by researchers to meet clients' needs. If you have needs in LNT production, please contact us directly.
Reference
- Özcan, E.; Sela, D.A. Inefficient metabolism of the human milk oligosaccharides lacto-N-tetraose and lacto-N-neotetraose shifts Bifidobacterium longum subsp. infantis physiology. Frontiers in nutrition. 2018, 5: 46. (Open Access)
