Study On The Preparation,Purification And Biological Activity Of Sulfated Scallop Viscera Oligosaccharide

The scallop is an important aquatic economic species. The processing by-products, scallop viscera, are often discarded, which cause heavy economic losses and environmental pollution. To improve the use value of scallop viscera, the polysaccharide from the viscera of scallop (Patinopecten yessoensis) were extracted in this study. The polysaccharides were degradated and modified by chlorosulfonic acid-methanamide method. The antioxidant, antitumor and immunoregulatory activities of the sulfated oligosaccharides obtained were tested, which lay the foundation for comprehensive development and utilization of the processing by-products of scallop.Crude scallop viscera polysaccharide (CSVP) was obtained from scallop viscera by proteolytic extraction in this study. The proteins of CSVP were removed by protease-sevag method and freezing-thaw method, and scallop viscera polysaccharide (SVP) was obtained. SVP was degradated and modified by chlorosulfonic acid-methanamide method, and sulfated scallop viscera oligosaccharide (SSVO) was obtained. Infrared spectrum analysis indicates that SSVO have characteristic absorption of sugar and sulfate. SSVO was fractionated by gel-filtration chromatography with Bio-Gel P-6, giving two fractions termed SSVO-Ⅰ and SSVO-Ⅱ. The molecular weight (MW) for SSVO-Ⅰ and SSVO-Ⅱ are5900Da and2400Da, respectively. The degree of sulfatation (DS) for SSVO-Ⅰ and SSVO-Ⅱ were of0.98and0.86, respectively. The monosaccharide composition of SSVO-Ⅰ is Rha, Fuc, Ala, Xyl, Man, Gal and Glu in a molar ratio of1.0:1.3:2.4:3.0:1.3:4.1:2.5. SSVO-II has the same monosaccharide composition with corresponding molar ratio of1.0:1.5:2.1:3.4:1.6:2.5:2.4.The antioxidant activities were tested in vitro, indicating SVP、SSVO-Ⅰ and SSVO-Ⅱ possess DPPH free radicals scavenging activity, hydroxyl free radicals scavenging activity and metal ions chelating activity. The potency of scavenging DPPH free radicals is that SSVO-Ⅰ>SSVO-Ⅱ; The potency of scavenging hydroxy free radicals is that SVP> SSVO-Ⅰ>SSVO-Ⅱ; The potency of metal ions chelating is that SSVO-Ⅰ>SSVO-Ⅱ>SVP. Relatively speaking, the antioxidant activity of SSVO-Ⅰ is stronger than that of SSVO-Ⅱ in vitro. Antitumor, immunomodulating and anticoagulating activity of SVP, SSVO-I and SSVO-II were investigated in vitro. Results showed that SSVO-Ⅰ could restrain the proliferation of Hela with a inhibitory rate of24.25%at a final concentration of1000μg/mL. The inhibition effect of SSVO-Ⅱ and SVP to Hela was not obvious. SSVO-I and SSVO-Ⅱ could significantly enhance lymphocyte proliferation and peritoneal macrophage phagocytosis activity, could also markedly prolong APTT (Activated partial thromboplastin time), PT (prothrombin time) and TT (thrombin time), suggesting potential immunomodulating and anti-intrinsic coagulating function and application value.