Empowering Health Through Advanced Capsular Polysaccharide (CPS) Production
CPS are large molecules formed by the polymerization of 2 or more monosaccharides and other substances (e.g. phosphate, sialic acid, etc.) as repeating units. Most bacterial CPSs are acidic mucopolysaccharides and belong to the heteropolysaccharide group.
As a company engaged in research in glycobiology, CD BioGlyco offers a wide range of carbohydrate production services to clients. Examples include Marine Carbohydrate Production Services. Here, we provide high-quality CPS production service to our clients with first-class Technologies for Carbohydrate Production and Purification.
Isolation and extraction of CPS
- Bacteriophage removal and crude extraction of total sugars
Before extracting CPSs, it is necessary to remove the bacterium first. At present, the main methods to remove the bacterial body to extract total sugar include centrifugation, enzymatic digestion, pasteurization, etc.
- Centrifugation. The sterilized culture solution is centrifuged and the supernatant is collected to remove the organisms.
- Enzymatic method. For CPSs, which contain unstable structures such as salivary acid, the most commonly used method is enzymatic digestion, i.e., centrifugation of the bacterial culture to obtain the organisms, then dispersing the organisms in buffer, then adding different enzymes to enzymatically digest the organisms, and finally centrifuging again to separate the fragments of the organisms from the CPS.
- Pasteurization. By heating and sterilizing the bacterial liquid, the solubility of polysaccharides is gradually increased with the increase in temperature. This method has the advantages of simple operation and better removal of bacteria.
- Removal of proteins and nucleic acids
Organic reagents cause the precipitation of polysaccharides and proteins. To avoid mixing nucleic acids and proteins in polysaccharides, we add cationic groups to the solution to which organic reagents are added and use the cations to combine with negatively charged groups to reduce the repulsion between polyribonucleotide chains, thus removing nucleic acids and proteins, and obtaining the precipitation of CPSs.
Purification of CPS
CPSs are substances with high polarity, different molecular weights, and different charge properties, so they can be separated and purified by volumetric exclusion chromatography and ion exchange chromatography.
- Gel chromatography
Gel chromatography is one of the most effective of the volumetric exclusion chromatography techniques. It is a simple and fast separation technique, which is classified into cross-linked dextran gel, agarose gel, acryl dextran gel, etc. depending on the type and nature of the gel. We select the appropriate gel for purification of CPSs according to the client's project requirements.
- Ion exchange chromatography
Ion exchange chromatography can effectively remove acidic and alkaline components and inorganic ions from CPSs, thus obtaining high-purity CPSs.
Characterization of CPS
- Monosaccharide composition
We analyze the monosaccharide composition of CPSs using high-performance liquid chromatography (HPLC), gas chromatography-mass spectrometry (GC-MS), and ion chromatography.
- Structural analysis
GC-MS is an effective tool for analyzing the structure of CPSs. Methylation, hydrolysis, reduction, and acetylation of CPSs are performed sequentially, and then the positions and sequences of the sugar residues are determined by GC-MS. In addition, nuclear magnetic resonance (NMR) technology is also a good method to identify the structure of CPS. NMR profiling can be used to deduce the location of carbon chain linkages, conformation, and other information in the CPS, as well as to identify the samples.
Publication Data
Technology: Ethanol precipitation, hexadecyltrimethylammonium bromide precipitation
Journal: Frontiers in Bioengineering and Biotechnology
IF: 5.7
Published: 2018
Results: In this article, the authors describe a method for the purification of CPSs that applies to 16 bacterial CPS types. The method involves the steps of ethanol precipitation, hexadecyltrimethylammonium bromide precipitation, and activated carbon purification. Experimental results showed that the method was able to remove more than 99% of contaminating proteins, nucleic acids, and carbohydrates while retaining the immunogenicity of the CPSs.
Applications
- CPS acts as a virulence factor and plays an important role in the resistance of bacteria to phagocytosis by phagocytes. Therefore, the anti-phagocytic ability of CPS could lead to the development of vaccines against infections caused by the respective pathogens.
- CPS promotes bacterial-to-bacterial or cell-to-cell adhesion and can be used to promote biofilm formation and maintain cell wall morphology.
- When nutrients are lacking, CPSs can be used as a carbon and energy source to provide energy to bacteria.
Advantages
- Our state-of-the-art marine carbohydrate extraction and purification technology ensures efficient isolation and purification of carbohydrates from a variety of marine sources.
- Our CPS production line removes impurities through various purification techniques to ensure product purity.
- We provide tailor-made CPS production solutions according to the needs of our clients' projects and offer detailed data interpretation services.
Frequently Asked Questions
What are the biological functions of CPSs?
Desiccation tolerance. Capsular is highly hydrated and CPS enhances bacterial dehydration tolerance by forming a physical barrier that retains moisture.
Frost resistance. CPS inhibits ice recrystallization by pinning and fixing ice particle boundaries, thereby protecting the bacteria at low temperatures.
Adhesion. Certain bacterial CPSs selectively bind to specific cell surfaces, rendering CPS adhesive.
Resistance to cationic antimicrobial peptides. Negatively charged CPSs can be used as bait to bind positively charged cationic antimicrobial peptides, reducing the amount of antimicrobial peptide reaching the surface of the pathogenic bacteria and preventing ionic flux disturbances on both sides of the membrane due to the disruption of the structure of the cytoplasmic membrane of the pathogenic bacteria by the antimicrobial peptide.
How do CPSs help bacteria?
CPSs are one of the keys to the adaptation of bacteria to their environment, covering the cells of many different bacterial species. CPS acts as a magical cloak that protects the bacteria from toxic compounds and desiccation, allowing them to attach themselves to the surface of objects and evade the host's immune system.
At CD BioGlyco, our service team has a deep background in Marine Biomolecule research and is highly recognized in the field. We help our clients advance their CPS research projects with our highly skilled marine microbial carbohydrate production technology. Please feel free to contact us if you have any needs or difficulties in the field of CPS, and our staff will reply to you promptly.
Reference
- Morais, V.; et al. Purification of capsular polysaccharides of Streptococcus pneumoniae: traditional and new methods. Front. Bioeng. Biotechnol. 2018, 6: 145.
