摘要
本文对发芽糙米粗多糖进行纯化,通过比较活性炭吸附法、过氧化氢氧化法、大孔树脂吸附法三种方法的脱色效果,以及Sevage法、三氯乙酸法、酶与Sevage结合方法三种方法的脱蛋白效果,筛选出发芽糙米粗多糖脱色、脱蛋白的最佳方法。分别比较了DEAE-Sepharose CL-6B、DEAE Fast Flow、DEAE Sepharose 52三种柱层析填料对糙米粗多糖的层析纯化效果,筛选出最优填料。并对纯化后的发芽糙米多糖各组分进行分子量的测定。结果表明:大孔树脂AB-8对发芽糙米粗多糖脱色效果最佳,脱色率为86.57%,多糖损失率为28.96%。酶-Sevage法脱蛋白效果较好,且多糖损失率低,脱蛋白率为74.36%,多糖损失率为14.09%。对发芽糙米多糖进行柱层析的最佳填料为DEAE-Sepharose CL-6B。根据线性回归方程计算其平均分子量,水洗多糖组分的平均分子量为1.47×10~5Da,盐洗多糖组分的平均分子量分别为9.62×10~5、5.59×10~6、3.15×10~5Da。结论:确定了针对发芽糙米粗多糖的去除蛋白质和脱色的方法,并筛选出最佳的柱层析填料,分离出四个组分发芽糙米多糖,为发芽糙米粗多糖的提取、纯化、分离逐渐转变为工业化生产提供了理论基础。
In this paper,the crude polysaccharides from germinated brown rice were purified. The decolorization effects of activated carbon adsorption,hydrogen peroxide oxidation and macroporous resin adsorption were compared.The Sevage method,trichloroacetic acid method,enzyme and Sevage method were compared. These method was to screen out the best method for decolorization and deproteinization of germinated brown rice polysaccharide.The chromatographic purification effects of DEAESepharose CL-6B,DEAE Fast Flow and DEAE Sepharose 52 on crude polysaccharides from brown rice were compared and the optimal fillers were screened.The molecular weight of each component of the purified germinated brown rice polysaccharide was measured.The results showed that macroporous resin AB-8 had the best decolorization effect on germinated brown rice polysaccharide,the decolorization rate was 86.57%,and the polysaccharide loss rate was 28.96%.The enzyme-Sevage method had good deproteinization effect and low polysaccharide loss rate,the deproteinization rate was 74.36%,and the polysaccharide loss rate was 14.09%.The best filler for column chromatography of germinated brown rice polysaccharide was DEAE-Sepharose CL-6B.The weight average molecular weight was calculated according to the linear regression equation.The average molecular weight of the washed polysaccharide component was 1.47 × 10~5 Da,and the average molecular weight of the salt washed polysaccharide component was 9.62 × 10~5,5.59 × 10~6,3.15 × 10~5 Da,respectively.Conclusion: The method for removing protein and decolorization of crude polysaccharide from germinated brown rice was determined,and the best column chromatography packing was selected.Four components of germinated brown rice polysaccharide were isolated to extract,purify and separate the crude polysaccharide from germinated brown rice,which provided a theoretical basis for the transformation of polysaccharides into industrial production.
引文
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