摘要
目的:优选铁皮石斛总多糖的胰蛋白酶联合Sevage法脱蛋白工艺,为该有效部位的深度开发和综合利用提供参考。方法:以总多糖保存率和蛋白脱除率为评价指标,通过单因素试验比较Sevage法、木瓜蛋白酶法、胰蛋白酶法和胰蛋白酶联合Sevage法;以蛋白脱除率为指标,酶解时间、酶用量、酶解温度和粗多糖溶液与Sevage试剂体积比为考察因素,利用BoxBehnken响应面法优化条件。结果:胰蛋白酶联合Sevage法具有较高的总多糖保存率及蛋白脱除率;最佳工艺条件为酶解时间1.5 h,酶用量0.21 g·m L-1,酶解温度63.25℃,总多糖溶液-Sevage试剂(4.27∶1);蛋白脱除率(87.15±7.93)%,总多糖保存率(81.32±8.54)%。结论:不同方法对铁皮石斛总多糖的脱蛋白效果有明显差异,胰蛋白酶联用Sevage法的效果最佳。
Objective： To optimize deproteinization technology of Dendrobii Officinalis Caulis.Method： With retention rate of total polysaccharides and deproteinized rate as the indexes,single factor tests were adopted to compare Sevage method, papain enzymatic, trypsin method and trypsin combined Sevage method.Effects of enzymolysis time,enzyme dosage,enzymolysis temperature and volume ratio of coarse polysaccharide solution to Sevage reagent on deproteinization technology was optimized by response surface method. Result：Trypsin combined Sevage method had higher retention rate of total polysaccharides and deproteinized rate; optimum deproteinization technology was as follows： enzymolysis time of 1. 5 h, enzyme dosage of 0. 21 g·m L^-1,enzymolysis temperature at 63. 25 ℃, coarse polysaccharide solution-Sevage reagent（ 4. 27 ∶ 1）. Under these conditions,deproteinized rate and retention rate of total polysaccharides were（ 87. 15 ± 7. 93） % and（ 81. 32 ±8. 54） %. Conclusion： Effect on total polysaccharides from Dendrobii Officinalis Caulis has significant difference with different deproteinized method,Trypsin combined Sevage method has optimum deproteinized effect.
出处
《中国实验方剂学杂志》
CAS
CSCD
北大核心
2016年第17期19-22,共4页
Chinese Journal of Experimental Traditional Medical Formulae
基金
湖南省教育厅重点项目(14A109)
湖南省科技厅发改委项目(湘发改高技[2013]1199号)
湖南省科技厅科研条件创新专项(2012TT2012)
湖南中医药大学校级课题(99820001134)
关键词
铁皮石斛
总多糖
脱蛋白
Sevage试剂
保留率
Dendrobii Officinalis Caulis
total polysaccharides
deproteinization
Sevage reagent
retention rate
