Following the enzymatic digestion of chondroitin sulfate by a simple GPC analysis

Silva, Carla; Novoa-Carballal, Ramón; Reis, Rui L.; Pashkuleva, Iva

doi:10.1016/j.aca.2015.05.034

Cited by 22 publications

(12 citation statements)

References 25 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…GAGs digestion : 10 mg of precipitated GAG were resuspended in 300 µL of PBS; 30 µL of 5 U/mL chondroitinase ABC (Sigma) were then added and enzymatic digestion was performed at 37 °C during 24 or 72 h. At each endpoint GAG were separated from the enzyme as described by Silva et al [ 64 ]; briefly: 150 µL of the GAGs/enzyme mix were heated for 25 min at 70 °C and then centrifuged at 12,000 g for 25 min. The pellet was discarded while supernatant containing digested GAGs was recovered, freeze-dried and kept for further analysis.…”

Section: Methodsmentioning

confidence: 99%

Bioinspiring Chondrosia reniformis (Nardo, 1847) Collagen-Based Hydrogel: A New Extraction Method to Obtain a Sticky and Self-Healing Collagenous Material

et al. 2017

View full text Add to dashboard Cite

Collagen is a natural and abundant polymer that serves multiple functions in both invertebrates and vertebrates. As collagen is the natural scaffolding for cells, collagen-based hydrogels are regarded as ideal materials for tissue engineering applications since they can mimic the natural cellular microenvironment. Chondrosia reniformis is a marine demosponge particularly rich in collagen, characterized by the presence of labile interfibrillar crosslinks similarly to those described in the mutable collagenous tissues (MCTs) of echinoderms. As a result single fibrils can be isolated using calcium-chelating and disulphide-reducing chemicals. In the present work we firstly describe a new extraction method that directly produces a highly hydrated hydrogel with interesting self-healing properties. The materials obtained were then biochemically and rheologically characterized. Our investigation has shown that the developed extraction procedure is able to extract collagen as well as other proteins and Glycosaminoglycans (GAG)-like molecules that give the collagenous hydrogel interesting and new rheological properties when compared to other described collagenous materials. The present work motivates further in-depth investigations towards the development of a new class of injectable collagenous hydrogels with tailored specifications.

show abstract

Section: Methodsmentioning

confidence: 99%

Bioinspiring Chondrosia reniformis (Nardo, 1847) Collagen-Based Hydrogel: A New Extraction Method to Obtain a Sticky and Self-Healing Collagenous Material

et al. 2017

View full text Add to dashboard Cite

show abstract

“…The former is evidence of a H4 proton chemical shift consistent with glyosidic bond cleavage and the generation of C4,5 double bonds (⌬ 4,5 ) in the terminal nonreducing end GlcA or uronic acid (UA) residue (⌬ 4,5 UA) of various products, whereas the latter peak at 5.2 ppm corresponds to resonance signals of the H1 anomeric proton of the same sugar ( Fig. 2D) (23)(24)(25)(26).…”

Section: Activity Of Btcdhmentioning

confidence: 95%

The human gut microbe Bacteroides thetaiotaomicron encodes the founding member of a novel glycosaminoglycan-degrading polysaccharide lyase family PL29

Ndeh

Muñoz‐Muñoz

Cartmell

et al. 2018

Journal of Biological Chemistry

View full text Add to dashboard Cite

Edited by Chris WhitfieldGlycosaminoglycans (GAGs) and GAG-degrading enzymes have wide-ranging applications in the medical and biotechnological industries. The former are also an important nutrient source for select species of the human gut microbiota (HGM), a key player in host-microbial interactions. How GAGs are metabolized by the HGM is therefore of interest and has been extensively investigated in the model human gut microbe Bacteroides thetaiotaomicron. The presence of as-yet uncharacterized GAG-inducible genes in its genome and of related species, however, is testament to our incomplete understanding of this process. Nevertheless, it presents a potential opportunity for the discovery of additional GAG-degrading enzymes. Here, we investigated a gene of unknown function (BT_3328) from the chondroitin sulfate (CS) utilization locus of B. thetaiotaomicron. NMR and UV spectroscopic assays revealed that it encodes a novel polysaccharide lyase (PL), hereafter referred to as BtCDH, reflecting its source (B. thetaiotaomicron (Bt)) and its ability to degrade the GAGs CS, dermatan sulfate (DS), and hyaluronic acid (HA). When incubated with HA, BtCDH generated a series of unsaturated HA sugars, including ⌬ 4,5 UA-GlcNAc, ⌬ 4,5 UA-GlcNAc-GlcA-GlcNac, ⌬ 4,5 UA-[GlcNAc-GlcA] 2 -GlcNac, and ⌬ 4,5 UA-[GlcNAc-GlcA] 3 -GlcNac, as end products and hence was classed as endo-acting. A combination of genetic and biochemical assays revealed that BtCDH localizes to the cell surface of B. thetaiotaomicron where it enables extracellular GAG degradation. BtCDH homologs were also detected in several other HGM species, and we therefore propose that it represents the founding member of a new polysaccharide lyase family (PL29). The current discovery also contributes new insights into CS metabolism by the HGM.

show abstract

“…The samples were digested with chondroitinase ABC and analyzed following the GPC method recently described by Silva et al (2015), to assure that 99% of the material was converted into oligosaccharides. A shift of the refractive index signal to elution times corresponding to pullulan disaccharides after 48 h of digestion is observed in the eluograms (Figure 4 and S4).…”

Section: Ch3 Galnacmentioning

confidence: 99%

By-products of Scyliorhinus canicula, Prionace glauca and Raja clavata: A valuable source of predominantly 6S sulfated chondroitin sulfate

Novoa-Carballal

Pérez‐Martín²,

Blanco³

et al. 2017

Carbohydrate Polymers

Self Cite

View full text Add to dashboard Cite

Chondroitin sulfate (CS) was isolated from Scyliorhinus canicula (fin, head and skeleton), Prionace glauca (head), and Raja clavata (skeleton) by-products from fish processing industry using environmentally friendly processes. The molecular weight was determined by gel permeation chromatography and the sugar composition and sulfation position by NMR and SAX-HPLC after enzymatic digestion. The CSs showed a prevalent 6S GalNAc sulfation for the 3 species (4S/6S ratio lower than 1). A higher 6S sulfation was observed for P. glauca head and R. clavata skeleton (4S/6S ratio below 0.20) than for S. canicula (4S/6S ratio ca. 0.6). The existence of CS samples with such low 4S/6S ratio has only been observed before in a rare species of shark (Mitsukutina owatoni, globin shark). The good extraction yields achieved make S. canicula, P. glauca and R. clavata fish industry by-products a useful source of 6-sulfated chondroitin sulfate.

show abstract

Following the enzymatic digestion of chondroitin sulfate by a simple GPC analysis

Cited by 22 publications

References 25 publications

Bioinspiring Chondrosia reniformis (Nardo, 1847) Collagen-Based Hydrogel: A New Extraction Method to Obtain a Sticky and Self-Healing Collagenous Material

Bioinspiring Chondrosia reniformis (Nardo, 1847) Collagen-Based Hydrogel: A New Extraction Method to Obtain a Sticky and Self-Healing Collagenous Material

The human gut microbe Bacteroides thetaiotaomicron encodes the founding member of a novel glycosaminoglycan-degrading polysaccharide lyase family PL29

By-products of Scyliorhinus canicula, Prionace glauca and Raja clavata: A valuable source of predominantly 6S sulfated chondroitin sulfate

Contact Info

Product

Resources

About