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This study Agrocybe material, design single factor test and orthogonal test, select not only good to maintain the the Agrocybe polysaccharide (Agrocybe cylindracea Polysaccharide ACP) biological activity but also improve the tea mushroom polysaccharides yield optimum extraction technology portfolio. The different off a protein purified Chaxingu crude polysaccharide, in order to determine the best deproteinized process. Tea mushroom polysaccharides isolated by column chromatography, molecular weight determination of its separated components, derived the active good polysaccharide molecular weight distribution of the characteristics and laws applied to the production of a simple, rapid Agrocybe the polysaccharides quality evaluation standards. Provide a reference for the establishment of other fungal polysaccharides product quality evaluation system. Single factor experiment of a single factor extraction conditions impact on of Agrocybe polysaccharide yield and activity design, fixed other factors change the condition of a factor, polysaccharide extraction rate determination and polysaccharide identification of the activity test, to determine the factors most suitable conditions, and to provide a reference for the Next orthogonal experiment design basis. Selection factors are: extraction temperature, pH value, and the cells broken way, and several other factors. The results show that: the discussion by single factor extraction conditions, select the pH 7.0,7.5,8.0 as three orthogonal test pH, level. Select 70 ℃, 80 ℃, 90 ℃ and three levels of orthogonal test temperature. Microwave treatment 6 min, the power of 400 w; ultrasonic treatment for 30 min; freezing and thawing three times and three levels of orthogonal test cell disruption by this factor. 2 extraction process based on the single factor of the tea mushroom polysaccharides yield and activity affect the pre-test conclusions, do three factors and three levels orthogonal test L9 (33) orthogonal table design by the determination of the polysaccharide extraction rate and polysaccharide activity Identification of the test, to determine the best combination of extraction process. The results showed that: improve the tea mushroom polysaccharides yield optimal combination of technology: a temperature of 90 ° C, pH value of 7.5, cell disruption by freezing and thawing three times; improve the the Agrocybe polysaccharide content of optimal combination of technology: a temperature of 80 ℃, pH value of 7.5, the cell disruption ultrasonic treatment for 30 min; enhanced Agrocybe Polysaccharides anti-lipid oxidation activity of the optimum combination of: a temperature of 80 ° C, pH value of 7.5, cell disruption as freezing and thawing 3 times; promote the optimum combination of mouse macrophage NO production: a temperature of 80 ° C, pH value of 7.5, the cell disruption ultrasonic treatment for 30 min. The integrated multi Factors consideration, high temperatures and alkali polysaccharide biological activity is not conducive to maintaining the freeze-thaw processing to significantly improve the yield rate of polysaccharide, and can maintain a good biological activity of polysaccharides, but contrary to the simplified process, the principle of reducing costs, Therefore, based on the results of the orthogonal experiment, extraction temperature 80 ° C, pH value of 7.5, cell disruption ultrasonic 30 min for the best combination of extraction process. 3 different deproteinization process Agrocybe polysaccharide activity Sevag method, enzymatic Sevag method associated with trichloroacetic acid, trichloroacetic acid with Sevag method associated with the and other electrical point of Agrocybe polysaccharide deproteinized research. The to the polysaccharide save rate and protein removal rate indicators, five methods the deproteinized effect, and junior pure polysaccharide off protein activity test, to determine the best deproteinized process. The results show that: the Sevag method of enzyme and Sevag method associated with trichloroacetic acid, trichloroacetic acid with Sevag method associated with the, isoelectric point of the tea mushroom polysaccharides deproteinized. The results show that: the five deproteinized method for deproteinized effect sevag Law and activity retention of polysaccharides are the best. Single hold in terms of the oxidation activity from polysaccharide Lipid too, better deproteinized polysaccharide activity. The relationship of the polysaccharide molecular weight and activity of the extracted Agrocybe polysaccharide through a sephadex column and DEAE-52 anion exchange column, and make efficient analysis of the polysaccharide obtained by the separation, and dextran standards efficient Spectrum draw range of molecular weight distribution of the various components do than analysis, respectively; each component anti-lipid peroxidation test and promote macrophages produce NO in vitro activity tests, screening out the activity component, which to determine the molecular weight distribution of the range of activity the better the tea mushroom polysaccharides. Range of the polysaccharides molecular weight distribution of polysaccharide biological activity can be found on Agrocybe polysaccharide anti-lipid oxidation activity, molecular weight in the order of magnitude of 105 polysaccharide anti-lipid peroxidation activity, the molecular weight or too small, all affect its activity. For Agrocybe polysaccharide stimulate macrophages to produce NO activity, ACP2-3 activity worst is 5.50 × 104-3.16 × 105, molecular weight range, the activity of ACP2-4 Preferably, the molecular weight range is 7.59 × 105-4.90 × 106 . ACP2-3 molecular weight of the smallest one of four components, and ACP2-4 compared to the other components, and a relatively large molecular weight, thus can be introduced activity than the molecular weight of a large molecular weight Agrocybe polysaccharide stimulate macrophages to produce NO little strong, but the the polysaccharides molecular weight and stimulate macrophages to produce NO activity is positively related needs further study. According to the above results, it can be seen, the different molecular weight of the polysaccharide, are very different in its biological activity, therefore, in the industrial processes must pay attention to the choice of the polysaccharide processing, according to the uses of the polysaccharide, the design of appropriate processing technology, in order to ensure better able to extract the activity of the polysaccharide.
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