Preparation and anticoagulation activity of a chemically sulfated polysaccharide (s-gfb) obtained from grifola frondosa

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Abstract

The aim of this study was mainly to determine anticoagulant activity of a chemically sulfated polysaccharide (S-GFB), which was derived from water-insoluble polysaccharide of Grifola frondosa mycelia. Through 14-day seed culture, maximum mycelial biomass and polysaccharides, which appeared at about the 90th hour fermentation, were 7.9 and 1.3 g/L, respectively. The sulfur content in S-GFB was 17.7% (w/w). The molecular structure images of S-GFB and GFB were seen with atomic force microscopy. S-GFB showed particle lumps at 1 μg/mL, which was different from the long polysaccharide chain of GFB at 1 μg/mL. Anticoagulant activities of the S-GFB were investigated by studying the activated partial thromboplastin time (APTT), thrombin time (TT) and prothrombin time (PT) using human plasma. APTT and TT were effectively prolonged, but no clotting inhibition was observed in PT assay at different concentrations. It proved that S-GFB had an important effect on decompounding fibrin polymerization through the fibrinolytic assay. The potential practical applications of this paper exhibited two points as follows: First, it took less than 4 days to obtain the maximum mycelial biomass in Grifola frondosa mycelial fermentation. So this method, which shortened the period of fermentation, may be utilized on industrial production of G. frondosa mycelia. Second, the water-insoluble polysaccharide accounted for a considerable proportion of polysaccharide obtained from G. frondosa. In order to make full use of the water-insoluble polysaccharide, we transferred it to water-soluble polysaccharide with high biological activity by chlorosulfonic acid-hydrochloride method.