Studies On Separation, Purification, Structure And Function Of Enhancing Immunoregulation From Soybean Glycoprotein

A glycoprotein is a macromolecule composed of a protein and a carbohydrate (a sugar). The carbohydrate is usually attached to the protein in a posttranslational modification, at asparagine, hydroxylysine, hydroxyproline, serine, or threonine. Possible carbohydrates include glucose, glucosamine, galactose, galactosamine, mannose, fructose, and sialic acid. The process of addition of sugar chains is termed "glycosylation". It exists in animal, plant and some microorganism widely. In this paper, the studies on separation, purification, structure and function of enhancing immunoregulation from Soybean Glycoprotein were investigated, in order to discover the effect of sugar chain in soybean glycoprotein.Soybean glycoprotein was extracted by PBS buffer solution and NaCl solution at 4℃ overnight after crushing, dryness and degrease. Then deposited by (NH₄)₂SO₄. According to the effect factor of PBS pH, salt concentration and extracted ratio, a 23 factorial design of orthogonal rotation was performed to find the best conditions of extraction. The best conditions were: PBS pH, salt concentration, extracted ratio. The best obtain percent was: PBS pH6.8, salt concentration0.15 mol/l, extracted ratiol:10The deposition of the soybean glycoprotein was dissolved in distilled water, dialyzed with PBS buffer solution and concentrated by PEG-6000, then separated with DEAE-Cellulose 52 IEC. After being eluted by PBS buffer solution, PBS -0.1mol/l NaCl and PBS -0.2mol/l NaCl, the soybean glycoprotein was collected. At the second place, the soybean glycoprotein was purified on Sephadex G-200 SEC, and was collected. The purity was 83.01% and sugar content was 9.84%.The Coomassie Brilliant Blue R-250 Staining and Shiff Staining of the soybean glycoprotein proved that it was glycoprotein not mixture of protein and sugar. The molecular weight of the soybean glycoprotein was 61000Da as determined by SDS-PAGE. The CD spectra showed that the soybean glycoprotein was typical a-helix and Infrared absorption spectra showed a- glycosidic bond and β- alkali treatment showed O-Glycans. The sugar chain was separated on Sephadex G-50 SEC and derived for GC spectra. The results indicated that the soybean glycoprotein was composed of Rhamnose, Xylose,...