Isolation, purification and structural analysis of substance with immuno-modulative activity from black soybean [Glycin max(L.) Merr.]
Date Issued
2008
Date
2008
Author(s)
Chien, Ya-Mei
Abstract
Black-soybean [Glycine max (L.) Merr.] has been used extensively as a healthy food and a traditional Chinese medicine for promoting good health. The related therapeutically qualities including immunomodulative activity, inhibition of human leukemic U937 cell proliferation, inducement of cell differentiation and promotion of hematopoiesis were reported from it’s polysaccharide.o further investigate the immunomodulatory property of black-soybean polysaccharides, black-soybean powder was first dissolved in dichloromethane and ethanol in order to remove the lipid-soluble and low-molecule weight components, and then extracted by water at 70 oC. The water soluble crude extracts were collected, lyophilized, further fractionated by Sephadex LH 20, TSK HW-55F chromatographies and deproteinized by proteinase K. The fractions containing polysaccharides were detected by phenol-sulfuric acid and the immuno-stimulatory activities were conducted in vitro by mouse J774A. 1 macrophage cell line. he carbohydrate composition analysis of crude black soybean extract was analyzed by Gas Chromatography-Mass Spectroscopy (GC-MS). Our result suggests that the monosaccharide composition of G1, G1P1, and G2P1 all pinpointed mannose and galactose as the major components together with smaller components of other sugars; G2P2 are composed of glucose and galactose as major and minor mannose. With regard to imuno-modulative activities, the more mannoses the better the activity.hrough NMR, the structure of G2P2 is determinated as stachyose that contains one fructose, one glucose and two of galactose. The linkage of G2P2 was terminal- fructose, terminal-hexose and two of α-1,6-hexose that determinated by Gas Chromatohraphy-Mass Spectroscopy (GC-MS), and the data was similar to NMR. Comparing G2P2 and stachyose immuno-modulative activity we found that G2P2 had immuno-modulative activities depending on dose-dependent, but stachyose were bio-inactive. The reason for immuno-modulative activity was that G2P2 was contaminated by mannose residue that can interact with macrophage and induce the cytokines secreted.
Subjects
black soybean
polysaccharide
immuno-modulation
polysaccharide-protein complex
cytokines
SDGs
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