| Viili is a traditional fermented dairy product, which is originated in Finland from Northern Europe. Its extracellular polysaccharide (EPS) has great physiological activity of lowering blood cholesterol, anti-tumor, anti-fatigue. In this project, milk was used to ferment viili for extracting extracellular polysaccharidethis. Viili EPS was used todetect the polysaccharide molecular weight, monosaccharide components and glycosidic bond configuration, and its anti-fatigue ability in animals.Single-factor test is used to optimize the deproteinization from Viili EPSthe result is:t Sevag reagent was 1/3 volume, the volume ratio of chloroform to n-butyl alcohol was 3:1, the shake time was 25min,deproteinizaiton time is 2 time is the best. On the basis of single-factor test, in addition to protein content as index deproteinized effect on Viili EPS optimizing by orthogonal test of Sevag reagent, the volume ratio of chloroform to n-butyl alcohol, shake time.The result showed that Sevag reagent was 1/4 volume of Viili polysaccharide solution, the volume ratio of chloroform to n-butyl alcohol was 3:1, and the shake time was 25min.Using the high performance gel permeation chromatography to detect the molecular size of Viili EPS:crude polysaccharides contain five molecular weight of polysaccharides, and the descending order is 2017kD,1069kD 508.0kD 56.58kD,4.946kD, respectively. By using silica gel thin layer chromatography and gas chromatography to detect the monosaccharide composition of Viili EPS. The result is Viili EPS is composed of rhamnose, arabinose, xylose, mannose, glucose and galactose and the molar ratio is 3.84:10.37:10.08:8.79:33.23:33.70 and the galactose and glucose is the main component. In light of the result of infrared spectroscopy, Viili EPS is preliminary determined as an a-type furan polysaccharide.The anti-fatigue effects of viili EPS on ICR mice show that Viili EPS can extend the swimming time of mice, from 289.50s of blank group to 616.63s of high dose group; reduce post-exercised concentration of blood urea nitrogen, from 6.96noml/L of blank group to 5.41nmol/L of high dose group; increase reserves amount of muscle glycogen from 1.230mg/g of blank group to 1.383mg/g of high dose group and liver glycogen from 9.307mg/g of blank group to 9.740mg/g of high dose group; enhance the ability to remove serum lactic-acid in mice from 6.796nmol/L of blank group to 5.573nmol/L of high dose group. The results suggest that Viili EPS can improve exercise tolerance ability of test mices and exert good impact on fatigue of physiological and biochemical indexes between 50、100、200 mg/kg bw Viili EPS.The test substance during the anti-fatigue experiment the doses with the anti-fatigue effect gave a positive correlation. In addition, from organ tissue section of desected mice the heart, liver, spleen, kidney weighs among each group did not differ significantly, the observation of the sectioned cells show the architech of the cells remain no pathological change by Hematoxylin and eosin (H&E) staining, which prove that there is no cytotoxic side effects among Viili exopolysaccharide. |
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