Overview
The human gut microbiota (HGM) is a large microbial community that is vital for human health. HGM maintenance depends on its ability to utilize complex polysaccharides in the form of dietary and host polysaccharides as a nutritional source. The mechanism by which the community metabolizes these polysaccharides is therefore essential to understanding the ecology of the system and how it can be manipulated to benefit human health. Bacteroidetes are one of the two major phylum that control HGM. Up to 20% of the genome of these organisms is dedicated to complex glycan metabolism in the form of genes that mainly encode carbohydrate active enzymes (e.g., polysaccharide lyase (PLs), glycosidase, glycan transport and binding proteins, and carbohydrate sulfatase).
The human gut microbiota (HGM), which is essential for human health, utilizes complex polysaccharides as its main carbon source. Glycosaminoglycans are an important and highly preferred source of nutrition for HGM. The metabolic pathways of various glycosaminoglycan substrates remain unknown. There have been researches on the biochemical, genetic and structural anatomy of its genetic locus, the glycosaminoglycan arrangement in organisms metabolizing Bacteroides polymorpha. It was pointed out that two previously unknown surface glycan-binding proteins were found that promoted the introduction of glycosaminoglycans into the periplasm. The metabolic pathways of glycosaminoglycans are determined by different kinetics and genetic specificities of various periplasmic lyases.
Creative Proteomics, as the leader of bio-tech industry, devote to forge high quality targeted metabolomics service with advanced apparatus and professional research team. We provide high throughput, high precision and short time-consuming Glycosaminoglycan Degradation Analysis Service, which could be meet all the need of our customers, to assist your scientific research.
Advantages
- Short time-consuming
- High sensitivity and few detection restrictions
- High precision and good repeatability
- High throughput
- Customized service
Fig 1. Model of glycosaminoglycan degradation by PUL CS/DS/HA.
Service workflow
- Extract and purifying glycosphingolipid from cell membrane or serum
- Release that glycosphingolipid glycan with the glucosidase
- The total methylation of glycan
- UPLC-HILIC-FLD analysis based on 2-AB labeling
- Analysis of fully methylated glycans by time-of-flight mass spectrometry
Fig 2. Workflow for the discovery-based proteomics.
Sample requirement
- Serum/plasma: 500 μl/sample
Repeated freezing and thawing of samples must be avoided. The serum sample should be precipitated in the collection tube for 30 minutes at room temperature, then transported to the centrifuge tube and centrifuged at 8000 rpm for 5 minutes. After centrifugation, the supernatant was equally divided into a freezing tube of 500 uL / sample.
- Protein: 100 µg
- Anticoagulated blood (EDTA): 1 mL
Anticoagulants and preservatives must be added immediately after collection and then frozen at -80 °C.
- Urine: 1 ml/sample
Urine samples should be equally divided into centrifuge tubes with 1 mL per tube, each tube is added with 1/100 (w/v) sodium azide and stored at -80 °C.
- Animal tissues: 200 mg/sample
Samples should be frozen in liquid nitrogen immediately and then transported to -80 °C for storage after collected.
- Cells: ≥ 1 × 107/sample
Cytoactive should be terminated immediately to maintaining cell integrity.
- Feces: 500 mg/sample
In general, to assure enough sample to fulfill the whole project, the volume of the single sample need to be offered as much as possible. The remaining samples will be stored for one year free of charge and returned at any time if necessary. All samples need to be stored and transported at -80°C and try to avoid using surfactants (SDS, Triton-X) and inorganic salts.
Clinical samples are repeated in no less than 30 cases in a single group.
Animal samples are repeated in no less than 9 cases in a single group.
Report delivery
- Experimental procedure
- Parameters of HPLC and MS
- Raw data, chromatograms and mass spectra
- Metabolites quantification data
- Custom analysis report
Service cycle
- Sample testing: 5-10 working days
- Data analysis: 5-10 working days
Creative Proteomics metabolism analysis platform is committed to the all-around, reliable and accurate analysis service for a variety of target substances, which is suitable for life-science research, drug exploration, biological determination and other fields. We sincerely hope to cooperate with you to assistant your scientific research.
References
- J.A.D. Van, S. Clotet-Freixas, A.C. Hauschild, et al. Urinary proteomics links keratan sulfate degradation and lysosomal enzymes to early type 1 diabetes. PLoS One, 2020, 15(5):e0233639.
- W. Wang, L. Shi, Y. Qin, et al. Research and Application of Chondroitin Sulfate/Dermatan Sulfate-Degrading Enzymes. Front Cell Dev Biol, 2020, 8:560442.
- Z. Wang, K. Arnold, Y. Xu, et al. Quantitative analysis of heparan sulfate using isotopically labeled calibrants. Commun Biol, 2020, 3(1):425.
For Research Use Only. Not for use in diagnostic procedures.
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