Lacto-N-triose (LNT II) Production Service
Lactose N-trisaccharide II (LNT II) is the core structural unit of human milk oligosaccharide which has many potential nutritional values. Based on years of experience in providing services in the field of glycobiology, CD BioGlyco provides professional and high-quality services for LNT II production.
Existence of LNT II in the Human Body
As an important skeleton, LNT II has a wide range of application prospects that can be further synthesized into different human milk oligosaccharides (HMOs) as raw materials, such as lacto-N-tetraose and lacto-N-neotetraose. LNT II is a core structural unit of HMOs and it has attracted substantial attention for its multiple potential functions for infants.
LNT II is located on the cell surface as a glycolipid component normally. In the 1990s, there was a great interest in developing synthetic pathways to this type of oligosaccharide unit, because it usually occurs as a common trisaccharide unit in the lactotriglycosylceramide corresponding to the glycosphingolipids of the lactose series. The oligosaccharide components of glycosphingolipids have numerous roles in biological events such as cell-cell recognition.
A Novel Food Containing LNT II
At present, its organic chemical synthesis method has been developed, including multi-step operations such as protection, deprotection, and glycosylation. In comparison, enzymatic and fermentation techniques to produce LNT II are more attractive options due to their high efficiency and safety. Currently, a novel food containing a part of LNT II is expected to be applied in infant foods, beverages, table sweeteners, whole meal replacements for weight control, food supplements, foods for special medical purposes, and various other foods. This new type of food is also mainly composed of various human milk oligosaccharides, it is obtained through microbial fermentation.
Key Technologies
CD BioGlyco mainly provides chemical synthesis, chemical enzymatic, fermentation, and enzymic catalysis to realize the production of LNT II. Our researchers work out some Strategies for LNT II production, including but not limited to:
- Construction of an engineered E. coli cell factory to produce LNT II.
- Introducing heterologous glucanotransferase.
- To increase the supply of the precursor of LNT II, related pathway enzymes such as glucosamine synthase, deacetylase, and pyrophosphorylase, were modulated, optimized, and overexpressed in combination.
- Enzymic catalysis
- Directed evolution and site-specific saturation mutation of glycosidase for further engineering.
- Identification of transporter genes and inhibition or increase of corresponding gene expression.
- Enzyme Immobilization
- To enhance the reusability and operational stability of our enzymes, we employ advanced immobilization techniques. This includes both traditional carrier-based methods and innovative carrier-free approaches like cross-linked enzyme aggregates (CLEAs).
- Enzyme immobilization significantly lowers operational costs, improves long-term process stability, and contributes to the economic viability of large-scale LNT II production.
Science of Glycans, Perfected for You: LNT II Production
Our LNT II production service is a cornerstone of our glycan offerings. We provide comprehensive support, from small-scale research quantities to large-volume industrial production.
- Project Consulting and Design
Consult with our experts for an in-depth consultation to understand your specific needs, including desired purity, yield, and applications. We'll select the most appropriate production strategy based on your requirements:
- Enzymatic production: LNT II synthesis is catalyzed by specific glycosyltransferases, suitable for producing products of extremely high purity and precise structure. We precisely control enzyme concentration, pH, and temperature to maximize yield and efficiency.
- Microbial fermentation: High titers of LNT II are produced using customized, metabolically optimized E. coli or yeast strains. This method is highly efficient at an industrial scale and offers significant cost savings for large-scale production needs.
- Custom synthesis: For special projects or specific structural modifications, we offer custom LNT II synthesis services, generating LNT II derivatives or isotopically labeled variants for specific research applications.
- Substrate Sourcing and Preparation
We source premium-grade, abundant, and cost-efficient substrates—such as lactose derived from dairy by-products—and implement efficient preparation protocols for biocatalytic processes. This approach ensures starting material sustainability while enabling the production of cost-effective end products.
- Biocatalysis and Fermentation
Tailored to project scale and requirements, we leverage highly specific enzymatic reactions or large-scale fermentation using our engineered microbial cell factories. The process undergoes continuous monitoring to maintain optimal conditions for high-yield conversion.
- Purification and Isolation
Post-LNT II synthesis, multiple downstream purification techniques—including ultrafiltration, chromatography, and selective fermentation—are employed to eliminate impurities, unreacted substrates, and undesired byproducts, guaranteeing the final product's highest purity.
- Quality Control and Analysis
Advanced analytical tools like mass spectrometry (MS) and high-performance liquid chromatography (HPLC) are utilized to validate the synthesized product's structure, purity, and concentration, ensuring compliance with or exceeding required specifications.
- Final Product Delivery
High-purity LNT II is prepared per client specifications, encompassing lyophilization and packaging. We provide comprehensive documentation—including detailed analytical reports and certificates of analysis—to ensure transparent project completion.
Workflow
Publication Data
DOI: 10.1186/s13068-021-02050-5
Journal: Biotechnology for Biofuels
IF: 4.6
Published: 2021
Results: The article focuses on the production of LNT II, an important precursor for synthesizing HMOs, using an engineered E. coli cell factory. Researchers constructed a biosynthetic pathway in E. coli to produce LNT II from N-acetylglucosamine (GlcNAc), a renewable and low-cost chitin monomer. By introducing heterologous β-1,3-acetylglucosaminyltransferase (LgtA) and optimizing gene expression levels, they initially achieved an LNT II titer of 0.12 g L−1. Further metabolic engineering, including inactivation of lacZ and nanE genes to block competing pathways, and overexpression of pathway enzymes (NagA, GlmM, GlmU), significantly increased LNT II production. Ultimately, a maximum titer of 15.8 g L−1 was attained in a 3-L bioreactor through fine-tuning enzyme expression and optimizing feeding strategies. This study provides a novel and sustainable approach for producing high-value LNT II from GlcNAc feedstock.
Fig.1 Construction the biosynthesis pathway for LNT II production using GlcNAc substrate in E. coli. (Hu, et al., 2021)
Advantages of Us
Rapid Titer Enhancement
Through years of focused R&D in metabolic engineering, we have developed proprietary chassis strains that achieve exceptionally high LNT II titers. Our systematic optimization of gene expression strength and the elimination of hydrolytic enzymes significantly enhance product accumulation, translating directly into lower production costs and faster project completion for our clients.
Guaranteed High Purity
Our integrated synthesis and downstream processing pipelines are meticulously designed to yield LNT II with 98% purity. We employ advanced analytical methodologies to ensure the absence of structural isomers and related impurities.
Experienced Project Management
Our team comprises experts with over a decade of specialization in glycobiology and fermentation science. We provide continuous technical consultation, assisting you not only in production but also in formulating strategies for regulatory submission and product scale-up.
Applications
Nutraceuticals and Functional Food Development
Nutraceuticals and functional foods: As a powerful prebiotic, LNT II is incorporated into various functional foods, beverages, and dietary supplements aimed at supporting adult gut health.
Biomedical Research and Microbiome Studies
Biomedical research: High-purity LNT II is an essential tool for researchers studying the function of HMOs, the gut microbiome, and their roles in human health and disease.
Pharmaceutical Development and Therapeutic Agents
Pharmaceutical development: LNT II and its derivatives are being explored as potential therapeutic agents for various conditions, including inflammatory bowel disease and other gut-related disorders.
Frequently Asked Questions
Customer Review
"CD BioGlyco's LNT II production service exceeded our expectations. Their expertise was evident from our first discussion, and they delivered a high-quality product on time, allowing us to accelerate our product development timeline."
- Dr. J.
"The team at CD BioGlyco was a pleasure to work with. They were highly responsive to our questions and provided clear, detailed analytical reports that gave us complete confidence in the product's quality."
-Dr. T.
Associated Services
For projects requiring a deeper understanding of glycan structures, our advanced analysis services identify and characterize complex glycans.
In addition to LNT II, we specialize in the custom synthesis of a wide range of glycans, including complex HMOs and other oligosaccharides.
CD BioGlyco provides high-quality, high-precision HMO separation and profiling services. We have the confidence to be your research assistant in HMO research.
CD BioGlyco stands as a leading provider of high-quality LNT II production services. By combining advanced biocatalytic and microbial fermentation technologies with a commitment to excellence and customer support, we deliver a product that is pure, scalable, and cost-effective. For more information, project inquiries, or to discuss your specific LNT II requirements, please do not hesitate to contact us.
Reference
- Hu, D.; et al. Engineering Escherichia coli for highly efficient production of lacto-N-triose II from N-acetylglucosamine, the monomer of chitin. Biotechnology for Biofuels. 2021, 14(1): 198. (Open Access)
