Structural and immunological feature of rhamnogalacturonan I-rich polysaccharide from Korean persimmon vinegar

• The crude polysaccharide (KPV-0) was fractionated to identify the immunostimulatory structure.
• A purified polysaccharide fraction (KPV-I) showed a highest macrophage-stimulating activity.
• KPV-I seems to be composed three kinds of polysaccharide such as a RG-I, a mannan, and a linear glucan.
• A large amounts of RG-I is likely a crucial role in expression of the activity in persimmon vinegar.

The crude polysaccharide (KPV-0) isolated from Korean persimmon vinegar was fractionated using gel filtration chromatography to enhance the immunostimulatory activity and to identify the structural features of active fraction. Among three fractions, KPV-I obtained in a void volume, demonstrated the potent production of macrophage-stimulating mediators, including tumor necrosis factor-α, interleukin (IL)-6, IL-12, and nitric oxide. KPV-I showed a combined single peak with high molecular weight of 55,000 Da by high performance size exclusion chromatography. Component sugar analysis revealed that KPV-I contained mainly of arabinose, mannose, galactose, rhamnose and galacturonic acid. Single radial gel diffusion assay using β-glucosyl Yariv reagent showed that KPV-I contained arabinogalactan protein with 13.7%. Methylation analysis indicated that KPV-I contained 21 kinds of neutral glycosidic linkages, which seemed to be composed three kinds of polysaccharide; that is a rhamnogalacturonan-I (65–70%) derived from persimmon as a raw material, a mannan (20–25%) derived from fermentation-associated microorganisms, and a linear glucans (less than 10%). In conclusion, polysaccharide isolated from persimmon vinegar could augment the macrophage stimulation, and a large amounts of RG-I polysaccharide derived from persimmon is likely a crucial role in expression of the activity in persimmon vinegar.

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