Isolation and Characterization of Low Molecular Weight Glycosaminoglycans from Marine Mollusc Amussium pleuronectus (Linne) using Chromatography

Abstract

The glycosaminoglycan (GAG) heparin is a polyanionic sulfated polysaccharide most recognized for its anticoagulant activity. In the present study, the GAGs were extracted from bivalve mollusc Amussium pleuronectus. The crude GAGs were fractionated by ion-exchange (DEAE-cellulose and Amberlite IRA-900 & 120) chromatography. The recovered active fractions (as determined by metachromatic assay) were confirmed by agarose gel electrophoresis and the active fractions were purified in Sephadex G-100 column. Fractionated and purified GAG molecular weight was determined through gradient polyacrylamide gel electrophoresis. The structural characterization of low molecular weight GAG was analyzed by Fourier transform infrared spectroscopy. The activated partial thromboplastin time of purified GAG is 95 IU/mg and has molecular weight 6,500-7,500 Da. The disaccharide compositional analysis on the GAG sample was sulfated like porcine intestinal mucosal heparan sulfate, and it contains equivalent amount of uronic acid and hexosamine. The results of this study suggest that the GAG from A. pleuronectus could be an alternative source of heparin.

Isolation and characterization of anticoagulant compound from marine mollusc Donax faba (Gmelin, 1791) from Thazhanguda, Southeast Coast of India

“…Vidhyanandhini (2010) reported that the amount of crude GAGs was estimated as 9.85 gm/kg and after purification by using amberlite and barium acetate, the yield was found to be 33 and 148 mg/kg of K. opima. Saravanan and Shanmugam (2010) reported the amount of crude and purified GAG was estimated as 17.2 g/kg and 48 mg/kg of tissue in A. pleuronectus. The result of the present study clearly shows that the yield of heparin and heparin-like glycoasaminoglycans is higher and lower than those of previously reports.…”

“…In the present study, the anticoagulant GAGs from whole body tissue of D. faba crude and purified sample showed a major peaks at 3437. 15, 1639.49, 1251.8, 1197.79, 1128.36 and 1078.21 cm -1 which is said to be for the GAGs groups (Saravanan and Shanmugam, 2010 Besides the aforementioned, it could also have a very good anticoagulant activity as the anticoagulant compound from this species proves the possibility of its utilization as an additional potent source for the extraction of such anticoagulant compound since the anticoagulant activity of the presents isolated heparin and heparin like GAGs from D. faba was even more than that of many heparins obtained from commercial sources.…”

Marine Carbohydrate Based Therapeutics for Alzheimer Disease-Mini Review

“…GAGs are long un-branched hetero-polysaccharides, widespread in nature, occurring in a great variety of organisms exhibiting various physiological and biological functions [29][30][31]. Figure 1 Decade-wise increase of marine compound discovery.…”

“…79°46′ E), Tamil Nadu, India. The samples are defatted and processed to obtain crude glycosaminoglycan (GAG) [53]. The crude GAG is subjected to enzymatic depolymerization to obtain low molecular weight heparan sulfate (LMW-HS) and its purity was determined by PAGE and metachromatic activity.…”

Low molecular weight heparins for current and future uses: approaches for micro- and nano-particulate delivery

“…These studies provide the key for novel drug treatments for a range of diseases, including cancer as well as various inflammatory diseases (Lever & Clive, 2002). Although commercial, pharmaceutical heparin preparations are strictly obtained from mammalian sources, marine mollusk have recently been identified as a potent alternative source for extraction of several heparin and LMWH-like polymers with high anticoagulant activity (Saravanan & Shanmugam, 2010;Periyasamy et al, 2013). Heparins [both unfractionated (UFH) and low molecular weight (LMWH) derivatives] remain as the agents of choice for pharmacologic thromboprophylaxis, treatment of thromboembolism during pregnancy, with pediatric patients and in myocardial infarction, cardiovascular surgery, coronary angioplasty and numerous other conditions (Eldor, 2002;Pineo et al, 2004;Greer, 2005;Baldwin et al, 2014;Molinari et al, 2014).…”