铁皮石斛原球茎多糖提取及脱蛋白工艺研究

研究了铁皮石斛原球茎培养物中多糖提取工艺并进行了不同脱蛋白方法的比较。以提取温度、提取时间和料液比三因素进行正交实验,得到多糖最佳提取工艺为提取温度90℃、提取时间60 min、料液比1∶40(g/m L),多糖提取率78.51%。通过一系列单因素实验和正交优化实验,分析比较Sevage法、盐酸法、三氯乙酸法和木瓜蛋白酶法脱蛋白工艺。结果显示,Sevage法第5次洗脱效果最好,多糖质量分数43.81%,蛋白质质量分数9.67%;盐酸在体积分数25%时效果最好,多糖质量分数52.17%,蛋白质质量分数为4.82%;三氯乙酸质量浓度为0.03 g/m L时,多糖质量分数最高35.50%,蛋白质质量分数7.34%;木瓜蛋白酶法反应时间90 min、温度70℃、酶含量30 U时,多糖质量分数85.01%,蛋白质质量分数为5.89%,效果最为理想。

Extraction and deproteinization technology for polysaccharide from Dendrobium officinale protocorm

We addressed polysaccharide extraction technology from Dendrobium officinale protocorm and compared different deproteinization methods. We conducted orthogonal experiments with extraction temperature and time, and the ratio of solid and liquid as parameters. We found its optimum extraction conditions included extraction temperature of 90 ℃, extraction time of 60 minutes and solid/ liquid ratio of 1∶40 ( g/ mL) , and extraction efficiency could reach 78.51 %. We further compared Sevage, hydrochloricacid ( HCl) , trichloroacetic acid ( TCA ) and papain deproteinization technologies through a series of single element experiments and orthogonal optimization experiments. Experimental results show that: the fifth elution effect of Sevage is optimal,polysaccharide content of 43. 81% and protein content of 9. 67%. Elution effect is optimal, polysaccharide content of 52. 17% and protein content of 4. 82%, when HCL concentration is 25%. When TCA concentration is 0. 03 g/ mL, polysaccharide content is 35. 50 % and protein content is 7. 34 %. For papain method, optimum extraction conditions are enzymolysis time of 90 min, temperature of 70 ℃, and enzyme content of 30 U. The corresponding best effect is polysaccharide content of 85.01% and protein content of 5.89%.