| Octopus is a marine mollusk which widely distributed in the Bohai Sea, Yellow Sea, East China Sea and South China Sea waters. Polysaccharides and glycoproteins from Octopus were tested the immune regulation effect, which could increase the proliferation of mouse spleen cell and non-specific function of immunosuppression mice. In this paper, seventeen kinds of products were extracted from the naked body and wrist of Octopus ocellatus using different kinds of separation and purification methods. They were extracted from naked body (named WBP-la, WBP-lb, WBP-2a, WBP-2b) or wrist (named EBP-1a, EBP-1b, EBP-2a, EBP-2b) by cool water, extracted from naked body (named WFP-1a, WFP-1b, WFP-2a, WFP-2b) or wrist (named EFP-1a, EFP-1b, EFP-1c, EFP-2a, EFP-2b) by enzymolysis. The primary structure of EFP-1a and WFP-2b were determined by chemical and modern instruments analysis techniques. The product of EFP-2a and WFP-2b were modified by chemical methods.Polysaccharides from the naked body and wrist of Octopus ocellatus were extracted in order by distilled water, enzymolysis with combination of papain and trypsin digestion, and the physico-chemical properties of WBP, WFP, EBP and EFP were studied respectively. The weight average molecular weights of these polysaccharides were 162.6 kD,266.4 kD,139.1 kD and 127.9 kD. The method of 1-phenyl-3-methyl-5-pyrazolone pre-column derivatization high performance liquid chromatography (PMP-HPLC) proved that the purified product of EFP, named EFP-1a was only composed by glucose. The composition of WFP-2b and EFP-2a (the purified WFP and purified EFP) were complex, including GlcN, GalNAc, Man, GlcUA, Gal and Fuc.EFP and WFP were further purified by Q sepharose 4 F.F. and Sephacryl S300, and three kinds of pure polysaccharides EFP-1a,EFP-2a and WFP-2b were acquired and were proved to be higher purities by HPGPC, their molecular weights were estimated as 101.8 kD,6.7 kD and 28.2 kD, respectively. The sugar contents of them were 92.3%,17.0% and 17.0%. Protein contents of them were 1.9%,22.7% and 15.7%, respectively. Uronic acid contents of them were 19.9%,8.8% and 14.1% respectively. Sulfate contents of them were 3.3%,4.9% and 8.0% respectively. The primary structure of EFP-la was elucidated by chemical methods and spectroscopic techniques, such as acid hydrolysis, PMP-HPLC, methylation, HPLC,IR,UV,NMR and GC-MS. The results showed that EFP-la is mainly composed of a-1,4-glucose, containing little-1,2-Glc branches. WFP-2b was degraded by methods of partial acid hydrolysis and TLC, and then was isolated by Bio-Gel P6. Some oligosaccharide fragments were acquired and the structure of a tri-saccharide was elucidated by HPLC, MS and NMR. The results showed that the structure of tri-saccharide is aGlcUA1→4(βGalNAc1→3)Fuc, in which Fuc isα,βconfiguration of mutual conversion.Aimed to improved the medicinal value, EFP-1a and WFP-2b were further sulfated by sulfur trioxide pyridine complex (SO3-Py) method to get two kinds of derivatives named as EFP-1aSand WFP-2bS. It was investigated that the sulfate content was 33.3%.It was less reported about the research of the polysaccharides from Octopus ocellatus, which had many pharmaceutical activities. In this paper, seven kinds of products of polysaccharides from Octopus ocellatus were researched primarily, including the research of enzymatic extraction product EFP-1a and the modification of the enzymatic extraction product EFP-2a and cool water extraction product WFP-2b. The results have based for the further research about structure-activity relationship of polysaccharides from Octopus ocellatus, and also will be helpful for the research of polysaccharides from other marine animals.
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