Functions of Neu5Ac in Living Organisms

Neu5Ac is a common Salivary Acid found widely in nature in a variety of animals and microorganisms. It is a carboxylated nine-carbon monosaccharide that is used in the synthesis of other salivary acids or derivatives of salivary acids. Neu5Ac is usually located at the ends of cell surface glycoproteins or glycolipids. It is an integral part of the oligosaccharide chains of mucins, glycoproteins, glycolipids, etc., and occupies a terminal nonreducing position. It is attached to the inner and outer surfaces of cell membranes in a variety of ways.

The free Neu5Ac residue is activated and pumped into the Golgi lumen as CMP-Neu5Ac via reverse transporter proteins. Through several post-synthetic modifications, such as methylation, sulfation, etc., followed by attachment to sugar chains by sialyltransferases, sialic acid complexes with different functions are formed. Neu5Ac is involved in the maintenance of the structure of biomolecules, cell signaling recognition, and other physiological functions.

Neu5Ac-based Cell Surface Glycoengineering Service at CD BioGlyco

CD BioGlyco is experienced in Glycoengineering Research. We offer high-quality targeted introduction and modification of Neu5Ac through gene editing technology. Due to the multiple biological functions and wide range of applications of Neu5Ac, we have optimized our manufacturing process to produce high-quality Neu5Ac. We also provide accurate and efficient Neu5Ac content assays based on our comprehensive assay platform.

Fig.1 Neu5Ac-based cell surface glycoengineering service. (CD BioGlyco)

• Targeted introduction and modification of Neu5Ac

Neu5Ac synthesis in cells is regulated by a variety of enzymes. With our knowledge of Neu5Ac synthesis, we target the introduction of Neu5Ac by Gene Editing techniques, including gene overexpression and gene knockout. In addition to this, we alter the structure of Neu5Ac on the cell surface by chemical modification.

• Neu5Ac production services

Neu5Ac can be prepared by enzymatic synthesis, extraction from natural products, polysialic acid hydrolysis, chemical synthesis, whole-cell biocatalysis, etc. We mainly use the following three methods to synthesize Neu5Ac.

• The microbial production method does not require the addition of expensive precursor substances and can be produced directly using inexpensive substrates.
• Enzymatic synthesis of Neu5Ac can improve the substrate conversion rate and obtain a higher concentration of the product.
• The whole-cell biocatalytic synthesis of Neu5Ac is simple, and cost-effective, without the need for enzyme purification and the addition of cofactors. The preparation of whole-cell catalysts is easier and cheaper. Meanwhile, the cell membrane can protect the enzyme from the influence of the unfavorable external environment. It has the potential for industrialized production.

• Neu5Ac content analysis services

Neu5Ac determination is an important part of a biopharmaceutical development program. The structure and relative content of Neu5Ac need to be monitored during process development, production purification, stability testing, etc. We use liquid chromatography-mass spectrometry (LC-MS) and high-performance liquid chromatography (HPLC) to detect changes in Neu5Ac content in biologics.

Publication

Paper Title: Efficient production of N-acetylneuraminic acid in Escherichia coli based on the UDP-N-acetylglucosamine biosynthetic pathway

Technology: Gene Knockout, Gene Overexpression

Journal: Journal of Agricultural and Food Chemistry

IF: 6.1

Published: 2023

Results: In this study, several strategies were used to increase the production of Neu5Ac in E. coli BL21. The production of Neu5Ac was successfully increased by knocking down the expression of genes in the competing pathway. The result of overexpression of UDP-GlcNAc synthesis pathway-related enzyme genes showed that the UDP-GlcNAc pathway was enhanced. Finally, the results of the LC-MS assay showed a significant increase in the yield of Neu5Ac.

Fig.2 Engineered pathway for Neu5Ac biosynthesis in E. coli. Dashed, blue arrows, and red “X” indicate multiple reactions, gene overexpression, and gene knockout, respectively. (Zhao, et al., 2023)

Applications

• The biological function of Neu5Ac can be investigated by knockdown or overexpression of Neu5Ac synthesis-related gene
• High-quality Neu5Ac is used to study the biological functions of its, Neu5Ac derivatives, glycans, and glycoconjugates.
• Neu5Ac shows advantages in drug delivery systems for tumor targeting, and cancer inhibition. High-quality Neu5Ac products are used to study its application in drug delivery.
• Neu5Ac is widely used in the food industry. High-quality Neu5Ac is used to develop functional foods.

Advantages

• We are proficient in gene editing technologies such as CRISPR/Cas9, which enables precise editing of genes.
• We develop inexpensive production processes with high substrate conversion for Neu5Ac production.
• Efficient Neu5Ac production technology enables the synthesis of high-quality Neu5Ac, allowing researchers to efficiently study its application in medicine.
• For drug safety, efficacy, etc., the type and amount of Neu5Ac must be tested at all stages of product development. Neu5Ac analytical techniques help with quality control of drug development.

CD BioGlyco is a world leader in salivary acid glycoengineering. In addition to Neu5Ac, we also provide N-Glycolylneuraminic Acid and N-Glycolylneuraminic Acid Glycoengineering services. If you would like to know more about Neu5Ac's parameters, experimental program, etc., please feel free to contact us to inquire.