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Bailing is a precious Food and Medicine Fungi polysaccharides with anti-tumor, anti-aging, improve immune system function, and other effects. Bailing Mushroom strains submerged fermentation process, the mycelium ingredients, exopolysaccharide extraction, separation and purification of the extracellular polysaccharide, physicochemical properties and biological activity, a more systematic study for the development of Bailing mushroom fermentation products laid foundation. For the purpose, in order to obtain higher mycelial biomass by orthogonal experiment to determine the the Bailing optimal liquid fermentation medium composition: corn flour 3.0%, 1.0% glucose, 0.1% yeast extract, 2.0% of the soybean meal, KH2PO4 0.3%, MgSO4 · 7H2O 0.1%, CaSO4 0.1%. Studied in shake flask culture conditions test the pH, medium volume, inoculum size, rotation speed and other factors on biomass. The results show that the shake flask culture conditions: initial pH value of 6.5, medium volume 80ml/250ml, inoculum size 10% shaker at 160rpm fermentation cycle for 6d. In shake flask cultures on a trial basis, the fermentor test results show that: the use 5L fermentation tank, the ventilation volume (v / v) of 1:0.5, and the stirring speed was 100 r / min, and the liquid volume 3L under the conditions of a temperature of 25 ± 1 ℃, the mycelium biomass reached 2.9g/100ml. Analysis of the fermentation of Pleurotus mycelium and fruiting bodies ingredients: methanol extract of fermented mycelium and fruiting body, by ultraviolet scanning from the peak shape is roughly the same, a strong absorption at 210nm and 260nm at peak, both are similar in composition. The fermented mycelia in the amino acid composition consistent with the fruiting body and mycelium of total amino acids, essential amino acids were significantly higher than the fruiting body. Exopolysaccharide optimal extraction route is: the centrifugal removal of the mycelia of the fermentation broth was adjusted to pH 6.0,60 ° C and concentrated to 1/2 of the original volume of precooled 95% ethanol is added after cooling to a final concentration of 75 %, 4 ℃ freezer and chill for at least 24 hours. 5000rpm The precipitate was collected by centrifugation, the precipitate was dried over anhydrous ethanol, repeatedly washed with acetone to obtain the crude polysaccharides. Sevag reagents to remove protein, low pressure vacuum concentrated to a small volume, dialysis to remove small molecules, vacuum freeze-drying, was refined polysaccharide. Then dried DEAE-DE 52, and Sephadex G-100 column chromatography to give the pure product of the polysaccharide, the polysaccharide content of 90.5%, and gas chromatographic analysis showed: the polysaccharide by D-galactose, D-glucose, D-mannose, the D- xylose, D-arabinose and other components, the molar ratio of 2.448: 1.466: 0.684: 0.700: 0.586. The molecular weight of 2.79 × 105, the infrared spectrum measured obtaining β-type pyranose. Study of the Pleurotus biological activity by in vivo and in vitro experiments, the results show that: the extracellular polysaccharide has strong free radical scavenging ability to inhibit · OH-induced mouse liver tissue MDA formation and liver mitochondrial swelling in the concentration of 1mg/ml when they reach a significant level (P lt; 0.01); exopolysaccharide increase serum and liver tissue SOD, GSA and CAT activity and inhibition of MDA production capacity in lower concentrations can up to a significant level. The results showed that Pleurotus has strong biological activity in vitro and in vivo.
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