粒毛盘菌多糖脱蛋白方法及其条件优化

分别采用Sevage法、三氯乙酸-正丁醇(TCA-NBA)法、酶-Sevage法以及酶-TCA-NBA法对粒毛盘菌YM281胞外多糖进行脱蛋白。结果表明:用酶-TCA-NBA法对粒毛盘菌YM281胞外多糖脱蛋白的效果最佳,蛋白脱除率为51.57%,且此时多糖损失率(3.46%)为最低;进一步采用均匀设计优化该方法的脱蛋白条件,所得最优脱蛋白条件为:粒毛盘菌YM281多糖溶液20mL、木瓜蛋白酶溶液11mL、温度42.2℃、pH 4、酶解时间3.5h;除去变性蛋白后,再利用TCA-NBA法脱蛋白2次即可;采用所得的最优方法和条件脱蛋白,其蛋白脱除率高达72.3%,多糖损失率仅为2.93%。

Deproteinization method and optimal condition of crude polysaccharide from Lachnum sp.

Sevage method,TCA-NBA(trichloroacetic acid-N-butyl alcohol) method,enzyme-Sevage method and enzyme-TCA-NBA method were used to remove protein from extracellular polysaccharide of Lachnum YM281,respectively.The results showed that the method of enzyme-TCA-NBA was the most effective,by which the protein-removal rate and the polysaccharide-loss rate were 51.57% and 3.46%,respectively.Then the uniform design was employed to optimize the conditions of this method.As a result,the optimal conditions of the enzyme-TCA-NBA method were as follows:polysaccharide solution of Lachnum YM281 of 20 mL,papain solution of 11 mL,pH 4,zymolysis time of 3.5 h at 42.2 ℃;after removing the denatured protein,TCA-NBA method was used remove protein twice.Under this optimal method and conditions,the protein-removal rate reached up to 72.3% and the polysaccharide-loss rate was only 2.93%.