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KMID : 0545120080180030503
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Journal of Microbiology and Biotechnology
2008 Volume.18 No. 3 p.503 ~ p.511
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Evaluation of Biomolecular Interactions of Sulfated Polysaccharide Isolated from Grateloupia filicina on Blood Coagulation Factors
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Athukorala Yasantha
Jung Won-Kyo
Park Pyo-Jam
Lee Young-Jae
Kim Se-Kwon
Thava Vasanthan
No Hong-Kyoon
Jeon You-Jin
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Abstract
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An edible marine red alga, Grateloupia filicina, collected from Jeju Island of Korea was hydrolyzed by cheap food-grade carbohydrases (Viscozyme, Celluclast, AMG, Termamyl, and Ultraflo) to investigate its anticoagulant activity. Among the tested enzymatic extracts of G. filicina, a Termamyl extract showed the highest anticoagulant activity. Anion-exchange chromatography on DEAE-cellulose and gelpermeation chromatography on Sepharose-4B were used to purify the active polysaccharide from the crude polysaccharide fraction of G. filicina. The purified sulfated polysaccharide (0.42 sulfate/total sugar) showed ~1,357 kDa molecular mass and was comprised mainly of galactose (98%) and 1-2% of glucose. The sample showed potential anticoagulant activity on activated partial thromboplastin time (APTT) and thrombin time (TT) assays. The purified G. filicina anticoagulant (GFA) inhibited the coagulation factor X (92%), factor II (82%), and factor VII (68%) of the coagulation cascade, and the molecular interaction (protein-polysaccharide) was highly enhanced in the presence of ATIII (antithrombin III). The dissociation constant of polysaccharide towards serine proteins decreased in the order of FXa (58.9 nM) >FIIa (74.6 nM) >FVII (109.3 nM). The low/less cytotoxicity of the polysaccharide benefits its use in the pharmaceutical industry; however, further studies that would help us to elucidate the mechanism of its activity are needed.
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KEYWORD
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Grateloupia filicina, enzymatic hydrolysis, anticoagulant activity, ATIII, surface plasmon resonance
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