Empowering Research with Premium Peptidoglycan (PG) Production
PG is a major component of the cell wall, and its basic structure consists of a multimeric meshwork of N-acetylglucosamine and N-acetylmuramic acid linked by β-1,4 glycosidic bonds with tetrapeptide chains or peptide bridges between the repeating monomers. In biology, PG is not only essential for maintaining cell shape, size, and survival but is also the target of many commonly used antibiotics. As one of the world's leading biology companies, CD BioGlyco has in-depth experience in Marine Carbohydrate Production and long-term research in Marine Microbial Polysaccharide Production.
Here, with advanced Technologies for Carbohydrate Production and Purification, CD BioGlyco offers reliable and comprehensive PG production services to clients.
Extraction and purification of PG
- Ultrasonic method
PG is extracted from the cell wall of microorganisms by breaking the cell wall by the cavity effect of ultrasonic waves and then centrifuging to collect the precipitate. In this process, trypsin, trichloroacetic acid, and ether are used to remove proteins, phosphoric acid, and lipids from the cell wall to obtain a purer PG.
- Lysozyme method
This method uses Mycobacterium solani to hydrolyze glycosidic bonds in N-acetylglucosamine and N-acetylmuramic acid in the microbial cell wall, to fragment the microbial cell wall, and to collect the PG from the cell wall by centrifugation.
- Trichloroacetic acid method
PG is obtained by placing microorganisms in a trichloroacetic acid solution, after heating and centrifugation, and adding trypsin and ether to remove impurities such as proteins and lipids.
Characterization of PG
- Structural analysis
The structure of PG is examined using infrared spectroscopy (IR). The PG extracts are pressed with KBr and then scanned using an infrared spectrometer.
- Amino acid composition
High-performance liquid chromatography (HPLC) is an important tool for analyzing the amino acid composition of PG. We dry and pulverize the PG extracts and analyze them using HPLC. The results of amino acid composition can be obtained based on the comparison of the peak areas of each amino acid and the standard samples.
- Determination of molecular weight
We employ sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) to characterize the molecular weight of PG. The PG samples are heated and boiled and then subjected to an SDS-PAGE experiment. At the end of the experiment, the samples are stained and decolored and the results are observed.
Publication Data
Technology: Flow cytometry
Journal: Bioconjugate Chemistry
IF: 4.7
Published: 2024
Results: In this article, the authors isolated and extracted PG from fecal samples and analyzed its structure, combined with probes and metabolic labeling, using techniques such as flow cytometry. The experimental results show that PG isolation from mouse and human fecal samples is feasible, and these results create an opportunity to combine PG analysis with gut bacterial status. In addition, the authors demonstrated metabolic labeling of PG in living mice using synthetic PG analogs, i.e., these analogs could be successfully integrated into the PG scaffold.
Fig.1 In vivo analysis procedure of PG with azide labeling. (Ocius, et al., 2024)
Applications
- Since PG is an ideal target for recognition by the eukaryotic immune system, the production of PG can be used for the development of immune-enhancing agents.
- PG has anticancer activity and the production of PG can be used for the development of new highly effective and low toxicity antitumor drugs of great value.
- Since its biosynthesis can be inhibited by many antibiotics such as cycloserine, vancomycin, mycopeptides & penicillin, the production of PG can be used in research of antibiotic therapy.
Advantages
- Our service team has a strong background in marine biomolecule production research and offers custom PG production solutions to ensure that they meet the needs of our clients' projects.
- We have advanced technology for carbohydrate production, purification, and characterization, and provide one-stop solutions from PG extraction to analysis.
- We not only provide custom PG production experimental solutions to our clients but also provide detailed and professional data interpretation.
Frequently Asked Questions
How PG is biosynthesized?
The biosynthesis of peptidoglycan is divided into three stages.
- Synthesis of PG precursor UDP-MurNAc-pentapeptide. This is the first step in the biosynthesis of PG, which occurs in the cytoplasm. The precursor UDP-GlcNAc is catalyzed by specific amino acid ligases and alanine racemase to form the PG precursor UDP-MurNAc-pentapeptide.
- Transmembrane flip-flop. The second stage occurs at the cytoplasmic membrane. The PG precursor UDP-MurNAc-pentapeptide is attached to undecaprenyl pyrophosphate on the inner side of the cytoplasmic membrane by phosphotransferase and is flipped across the membrane to the outer side of the cytoplasmic membrane by flippase.
- Polymerization and cross-linking. The third phase occurs in the periplasm. PG precursors flipped to the outside of the cytoplasmic membrane polymerize into nascent PG chains in the presence of glycosyltransferase, and transpeptidase catalyzes the formation of cross-links between short PG chain peptides of adjacent PG chains to form a reticulum of PG.
What are the biological functions of PG?
- Immunological activity. PG is an immune-enhancing agent that exerts its immune function by inducing the release or expression of a variety of non-specific or specific immune factors.
- Anti-cancer activity. PG can exert anti-tumour effects by activating macrophages and inducing the body's immune response.
CD BioGlyco has mastered advanced carbohydrate production technology and has accumulated extensive experience in the production of PG. In addition, our production services cover carbohydrates from a variety of sources, such as marine plant sources, marine animal sources, marine microbial sources, and so on. Please feel free to contact us if you have any needs in the field of marine carbohydrate production.
Reference
- Ocius, K.L.; et al. Noninvasive analysis of peptidoglycan from living animals. Bioconjugate Chemistry. 2024, 35(4): 489-498.
