Sulfated glycosaminoglycans from crown‐of‐thorns Acanthaster planci – extraction and quantification analysis

Abstract

In this article, the novel inventive steps for the extraction and quantification of sulfated glycosaminoglycan (GAG) from Acanthaster planci starfish, generally known as crown-of-thorns (COT), are reported. Starfish have been implicated with collagenous distributions within their body anatomy, thus making it a prima facie fact searching for the possibility that GAGs can be isolated from COT. In this study, total-, N-, and O-sulfated GAGs were extracted from three anatomical regions of the COT (integument, internal tissue, and coelomic fluid) and comparison was made. The result showed that body region of COT seemed to contain higher amount of sulfated GAGs as opposed to the arm region (55.79 ± 0.65 μg/mg was the highest amount in the body extracted from its coelomic fluid and 32.28 ± 3.14 μg/mg was the highest amount in the arm extracted from its internal tissue). COT's integument and coelomic fluid from its body region possessed the highest total of sulfated GAGs content with no significant difference (P < 0.05) between the two. All GAGs from COT comprised a higher percentage of N-sulfated GAGs than its counterpart, the O-sulfated GAGs. When compared with a similar previous study that used sea cucumbers as the sulfated GAGs source, COT possessed more total sulfated GAGs content per milligram as compared with the sea cucumber generally. This result seems to unveil this marine species' advantage per se pertaining to GAGs extraction biomass applicability. Thus, COT could now be the better alternative source for production technology of total-, N-, and O-sulfated GAGs.

RNA Contaminates Glycosaminoglycans Extracted from Cells and Tissues

“…In some of the earliest publications describing GAG isolations, DNase and RNase were used to remove contaminating polynucleotides from the GAG extracts [29–31]. However, more recent publications no longer describe (a combination of) these endonucleases [23, 24, 32–38], which will result in the ample presence of RNA in GAG extracts. Of course, depending on the specific downstream application of the GAG extract and additional purification steps such as ion exchange chromatography, not all GAG extractions require RNA digestion [39–41].…”

“…when HS presence is concluded from azure A staining on agarose gels based on a “known” band position compared to DS and CS [54] and may therefore be wrongfully assigned. Since polynucleotides are often not considered as potential contaminants in GAG extracts, the complexation with cationic dyes that occurs in DMMB assays may result in overestimated GAG yields and wrong assignment of relative GAG compositions [23, 24, 32–38]. It is difficult to discern from the results of studies that lack controls for RNA content whether GAG extracts were contaminated by RNA.…”

A Rapid Quantitative Dye-Binding Method of Screening Glycosaminoglycans Presence in Medicinal Plants

“…Here, O. stamineus was shown to contain the highest total sulfated GAG as compared to other three plants. From the previous study that investigated extraction and quantification of GAG from Acanthaster planci starfish, the highest GAG amount was found in starfish's body coelomic fluid at 55.79±0.65 µg/mg [32]. By comparison with A. planci, O. stamineus has potential to give much higher GAG amount.…”

10.12980/jclm.3.2015j5-44

“…The dissociated pellet solution was divided into two wells of 96-well plate and absorbance was recorded at 656 nm. The standard plots were obtained by using chondroitin sulfate standard solution and values of the test samples were determined by using a linear equation of standard graph [15]. …”