铁皮石斛多糖酶法提取工艺研究及其对免疫功能的影响

Optimization of Polysaccharide Extraction Technology from Dendrobium officinale by Enzyme

目的建立铁皮石斛多糖的酶提取工艺并初步就其对免疫功能的影响进行研究。方法以铁皮石斛多糖的转移率为评价指标,通过单因素试验与正交试验,研究酶的种类、酶的添加量、酶解温度、酶解时间、料液比、pH值等因素对铁皮石斛多糖提取效率的影响;设空白对照组、模型对照组、匹多莫德组、铁皮石斛高、中、低剂量组(0.08mg·kg-1、0.04mg·kg-1、0.02mg·kg-1),连续灌胃给药30天。实验结束后进行脏器指数、碳廓清指数、吞噬指数等免疫指标检测。结果 1g铁皮石斛原料添加木瓜蛋白酶50U、中温α-淀粉酶100U,料液比为25∶1(m L∶g),酶解pH值为7、酶解时间为3h、酶解温度为55℃时,为最佳提取条件,铁皮石斛多糖的平均转移率为98.11%,优于已有文献报道;铁皮石斛多糖可以提高小鼠的脾脏指数、胸腺指数和碳粒廓清指数,但对吞噬指数无明显影响。结论建立了一种高效、温和的铁皮石斛多糖提取方法;提取的铁皮石斛多糖能增强免疫功能。

OBJECTIVE To set up enzyme extraction technology applied for extracting polysaccharides from Dendrobium officinale and study its effect on immune function. METHODS （1）Using the transfer rate of polysaceha- rides extracted from Dendrobium officinale as an evaluation index. The effect factors such as enzyme type, enzyme ad- dition amount, enzymolysis temperature, enzymolysis time, ratio of material and liquid and pH value on the extraction efficiency of Dendrobium officinale were studied by single factor test and orthogonal test. （2）50 mice was divided into 5 groups ： blank control group, model group, pidotimod group （0. 03mg· kg - 1 ）, polysaccharides high, medium or low dose groups（0. 08,0. 04,0. 02mg · kg-l）, continuous oral administration for 30 days. And then the immune indexes such as organ index,carbon clearance index and phagocytosis index were detected. RESULTS When 1 g of Den- drobium officinale was added with 50 U papain, 100 U mesophilic oL-amylase, ratio of material and liquid was 25：1 （mL： g）, enzymolysis pH value was 7, enzymatic hydrolysis was 3 h, enzymolysis temperature was 55℃, the average transfer rate of polysaccharides was 98.11%, which was superior to the literature recorded; Dendrobium candidum polysaccharide can enhance the mouse spleen index, thymus index, carbon clearance index, but no significant change in the phagocytic index that phagocytic phagocytosis had no significant effect. CONCLUSION An efficient and gentle method for the extraction of polysaccharides from Dendrobium officinale was established and Dendrobium candidum enhance immune function.