Enhancing Biotechnology Through Extracellular Polysaccharide (EPS) Production
Due to the special living environment, Marine Biomolecule, unlike terrestrial organisms, can produce many Marine Carbohydrates with novel structures and special action, and with remarkable pharmacological stability. Among them, marine microorganisms are an effective resource for obtaining polysaccharides with good properties. Moreover, the EPS of marine microbial has attracted much attention for its unique physicochemical properties, complex structure, and excellent biological activity.
Here, with superior Technologies for Carbohydrate Production and Purification, CD BioGlyco offers high-quality EPS production services to clients.
Extraction of EPS
We offer a variety of methods to extract EPS from microbial organisms, such as aqueous extraction, acid-base extraction, organic solvent extraction, and enzyme extraction. Most of the neutral EPS can be extracted with water, and the extraction of EPS by complex enzymes has the advantages of high extraction efficiency, mild conditions, and easy separation of polysaccharides from impurities.
Purification of EPS
Purification of microbial EPS is usually performed in two steps: removal of proteins and removal of small molecule impurities.
- Removal of proteins
We offer several methods to remove proteins from EPS, such as Sevag, trichloroacetic acid, neutral lead acetate, zinc acetate, and potassium ferricyanide method. The EPS crude product can be obtained by this step. In this process, we use ultraviolet (UV) spectroscopy to detect the protein content of EPS to ensure the complete removal of protein impurities.
- Removal of small molecules
The crude EPS product is dissolved in water or buffer, and the small molecules are removed by ion exchange chromatography (IEC), gel chromatography, and macroporous adsorbent resin to obtain the pure EPS product.
Characterization of EPS
- Structural analysis
We have a wide range of techniques for determining the structure of EPS, such as nuclear magnetic resonance hydrogen and carbon spectroscopy (1H and 13C NMR), infrared spectroscopy (IR), and more.
- 1H and 13C NMR. After dissolving EPS with a deuterated reagent, its structure is analyzed using NMR techniques such as 1H and 13C NMR. For the specific connection order of the polysaccharide structure, we use 2D NMR to determine it.
- IR. We analyze the EPS by scanning with an IR using the KBr press method. IR is capable of detecting typical polysaccharide characteristic structures.
- Molecular weight analysis
We use gel permeation chromatography (GPC) to detect the molecular weight of EPS. The EPS sample is injected into the GPC instrument and its molecular weight is calculated according to the regression equation of the standard curve.
- Monosaccharide composition
The monosaccharide composition of EPS can be determined by high-performance liquid chromatography (HPLC). EPS samples are derived and analyzed by HPLC, the signals of the peaks are detected using a UV detector, and finally, the monosaccharide composition of the EPS samples can be known based on the time of peak appearance and the peak area ratio.
Publication Data
Technology: Phenol sulfate method and gas chromatography (GC)
Journal: Frontiers in Microbiology
IF: 5.2
Published: 2023
Results: In this paper, the authors successfully extracted Microcystis EPS and determined the content and monosaccharide composition of Microcystis EPS. Among them, the authors used glucose as a standard sample to determine the polysaccharide content by phenol sulfate method and the monosaccharide composition of EPS by GC. The experimental results showed that the main monosaccharide components of Microcystis EPS included glucuronic acid, glucose, arabinose, and galactose. With the extension of incubation time, the EPS of different Microcystis produced different changes, e.g., the abundance of glucuronic acid increased in M. wesenbergii EPS, while the abundance of glucuronic acid decreased in M. aeruginosa EPS. On the other hand, the galacturonic acid concentration of both Microcystis strains increased with incubation time.
Fig.1 Diagram of the composition and content of monosaccharides in EPS. (Yang, et al., 2023)
Applications
- EPS has a good effect on enhancing the taste of food and preserving the freshness of food, thus the production of EPS is widely used in food development.
- Most EPS have biological functions such as anti-tumor, immunomodulation, anti-aging, anti-infection, etc., which can be widely used in the research and development process of new drugs in the pharmaceutical field.
- Because microbial EPS has the advantages of safety, non-toxicity, easy degradation, and no secondary pollution, EPS can be used as flocculants in the field of wastewater treatment.
Advantages
- We have mastered the advanced technology for the production of marine microbial polysaccharides, and our services are adapted to a large range of EPS production needs.
- We offer a one-stop EPS production process, including manufacturing, characterization, activity analysis, and more.
- We provide prompt and reliable EPS after-sales service to ensure your peace of mind.
Frequently Asked Questions
What are the factors affecting the biosynthetic process of microbial EPS?
During microbial EPS synthesis, genetic, physiological, and environmental factors affect the polysaccharide synthesis results, of which the environmental aspects include physical, chemical, and biological factors. Conditional factors such as temperature, pH, type of carbon and nitrogen source, inorganic salts, carbon-to-nitrogen ratio, and aeration of the medium also affect the synthesis efficiency of microbial EPS. Among them, the nutrition in the medium has a direct impact on the synthesis of EPS, and due to the different species of EPS-producing bacteria, there are differences in the nutrient content requirements in the medium suitable for their growth.
What are the research themes in the metabolic engineering of microbial EPS?
Research on metabolic engineering of microbial EPS synthesis can be categorized into two approaches:
- Improving microbial EPS production. Increasing EPS production by modulating the activity of key enzymes in the nucleotide synthesis pathway. This approach can be used to increase EPS production by, for example, knocking out genes involved in the synthesis pathway or by improving the performance of enzymes involved in the process.
- Altering the structure of EPS. The metabolic flow of carbon in metabolic engineering is used to change the structural characteristics of EPS such as their components, molecular weight distribution, and spatial conformation by culturing them in media with different types and contents of carbon sources.
CD BioGlyco has deep experience in the production of EPS and has a long-term track record of providing the highest quality products and services to our clients. We produce pure EPS through advanced manufacturing processes and perform comprehensive characterization. Our team is always ready to answer any of your questions and provide personalized advice and support. Please feel free to contact us for any of your needs.
Reference
- Yang, H.; et al. The extracellular polysaccharide determine the physico-chemical surface properties of Microcystis. Front. Microbiol. 2023, 14: 1285229.
