Study On Extraction, Purification And Immune Activity Of Glycoprotein From Abalone Visceral

Haliotis discus hannai Ino, a species of Gastropoda of Mollusks, is an importantfishery resource in China. Recently, with the development of abalone aquaculture andthe exploitation of deep processing products, the by-product of abalone about30%-40%in weight was processed into fishmeal or abandoned. It is not only causedenvironment pollution, but also leaded to waste of resources. There had been someresearch in ablone gonad polysaccharides and Sterols, their immune activity had beenimproved. And glycoprotein extracted from marine ainimals had attracted widespreadattention. Therefore, objective of this study was to dilute and purify glycoprotein fromhaliotis discus hannai Ino visceral, furthermore, investigated the functions on theimmunity regulative effect. The purpose was to provid scientific basic data forcomprehensive utilization of abalone visceral. Main results achived in this research asfollows:1. The Haliotis discus hannai Ino visceral was rich in protein and fat, poor inpolysaccharides, their contents were13.6%,5.1%,0.6%respectively.2. The optimized conditions of ultrasound-assisted glycoprotein extraction wasselected by Orthogonal analysis on the basis of single factor experiments. Theoptimized conditions of ultrasound-assisted extraction were as follows: NaClconcentration was0.2mol/L, the ratio of material to solvent was1:9, ultrasonic powerwas300w and ultrasonic time was40min. The extraction rate of glycoprotein in theextraction was69.7mg/g by this method, the extraction ratio increased by28.5%compared to the tradition method.3. The glycoproteins were precipitated by (NH4)2SO4and isolated byDEAE-Cellulose, Sephadex G-100, SephacrylS-300HR. Results showed ahomogeneous glycoprotein (AGP-Ⅲ) was obtained. The molecular weight of AGP-Ⅲwas estimated to be73.7kDa by SDS-PAGE. AGP-Ⅲ contained12.4%total sugar, 76.6%protein.4. The configuration analysis of purified Abalone visceral glycoprotein AGP-Ⅲshowed Monosaccharide composition of AGP-Ⅲwas Man, GlcN, Rha, GalNAc,Glc, Gal, Xyl and Fuc.17kinds of amino acids were determined in AGP-Ⅲ,and it’srich in Val、Glu and Gly. AGP-Ⅲ shared with a typical absorption property ofglycoprotein in a pyranose β-glycosidic and O-glycoside linkage. AGP-Ⅲ had typicalconfiguration of glycoprotein.5. The immune activity of Abalone visceral glycoprotein AGP was investigated.Medium dosage (50mg/kg) and high dosage(100mg/kg) Abalone visceralglycoprotein can promote immunological function both in celluar and non-specificimmunity in immunodepression mice. It’s suggesttted that Abalone visceralglycoprotein could enhance the immunity of mice.6. AGP-Ⅲ hadInhibition on Influecza Virus Neuraminidase in Vitro.