滇黄精多糖提取工艺优化及体外抗氧化和降血糖活性分析

Optimization of polysaccharide extraction process from Polygonatum kingianum Collett & Hemsl. and analysis of in vitro antioxidant and hypoglycemic activities

  • 摘要: 【目的】优化滇黄精多糖提取工艺,分析其体外抗氧化及降血糖活性,以期为滇黄精产品规模化开发提供参考依据。【方法】使用复合酶法,将木瓜蛋白酶复合纤维素酶按一定比例加入至滇黄精提取液中,采用单因素—响应面试验提取滇黄精多糖,优化滇黄精多糖在料液比、pH、复合酶添加量及酶解温度下的最佳提取条件,并评价最佳提取条件下得到滇黄精多糖的体外抗氧化活性和降血糖活性。【结果】滇黄精多糖的最佳提取工艺:料液比1∶23(g/mL)、pH 4.6、复合酶添加量14.5 g/L、酶解温度52 ℃,在此条件下获得滇黄精多糖提取率为22.27%,与理论值(22.31%)无显著差异(P>0.05);各因素对多糖提取率的影响大小为酶解温度>pH>料液比>复合酶添加量,4个因素对滇黄精多糖提取率均有极显著影响(P<0.01)。优化工艺提取得到的滇黄精多糖对2,2'-联氮-双-3-乙基苯并噻唑啉-6-磺酸(ABTS)阳离子自由基、1,1-二苯基-2-三硝基苯肼(DPPH)自由基、羟基自由基和超氧阴离子自由基表现出较强的清除作用,半抑制浓度(IC50)分别为0.26、0.54、1.07和0.59 mg/mL,有明显的铁离子还原能力,IC50为0.62 mg/mL;以阿卡波糖作阳性对照,当质量浓度为1.2 mg/mL时,滇黄精多糖对α-葡萄糖苷酶和α-淀粉酶的抑制率分别为35.65%和49.83%,表现出一定的降血糖能力。【结论】经单因素试验和响应面试验优化的提取工艺可提高滇黄精多糖提取率,得到的多糖具有较强的体外抗氧化及降血糖活性。

     

    Abstract: 【Objective】To optimize the extraction process of Polygonatum kingianum Collett & Hemsl. polysaccharide and analyze its in vitro antioxidant and hypoglycemic activities,with a view to providing reference basis for the large-scale development of Polygonatum kingianum Collett & Hemsl. products.【Method】Papain complex cellulase were added to the extract of Polygonatum kingianum Collett & Hemsl. in a certain proportion to extract Polygonatum kingianum Collett & Hemsl. polysaccharides in a single factor-response surface experiment,and optimize the optimal extraction conditions of Polygonatum kingianum Collett & Hemsl. polysaccharides in the solid-liquid ratio,pH,complex enzyme addition and the enzyme digestion temperature. At the same time,the in vitro antioxidant activity and hypoglycemic activity of Polygonatum kingianum Collett & Hemsl. polysaccharides obtained under these optimal extraction conditions were further evaluated.【Result】The optimal extraction process for the polysaccharide of Polygonatum kingianum Collett & Hemsl. was 1∶23(g/mL),pH 4.6,complex enzyme addition 14.5 g/L and enzyme digestion temperature 52 ℃,the extraction rate of polysaccharides of Polygonatum kingianum Collett & Hemsl. obtained under this condition was 22.27%,which was not significantly different from the theoretical value(22.31%)(P>0.05),and the influence of each factor on the polysaccharide yield was as follows:enzyme digestion temperature > pH > solid-liquid ratio > complex enzyme addition. The 4 factors had extremely significant effect on the polysaccharide extraction rate of Polygonatum kingianum Collett & Hemsl. (P<0.01). The Polygonatum kingianum Collett & Hemsl. polysaccharides obtained by this method showed vital scavenging effects on 2,2'-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid(ABTS)cation radicals,1,1-diphenyl-2-picrylhydrazyl(DPPH)radicals,hydroxyl radicals,and superoxide anion radicals,with semi-inhibitory concentration(IC50) values of 0.26,0.54,1.07,and 0.59 mg/mL,and showed obvious reducing iron ions capacity,with an IC50 of 0.62 mg/mL. Using acarbose as a positive control,when the mass concentration was 1.2 mg/mL,the inhibition rates of α -glucosidase and α-amylase by Polygonatum kingianum Collett & Hemsl. polysaccharides were 35.65% and 49.83% respectively, which showed certain hypoglycemic ability.【Conclusion】The process optimized by single factor and response surface experiments can improve the extraction rate of Polygonatum kingianum Collett & Hemsl. polysaccharides,and the polysaccharides obtained have strong in vitro antioxidant and hypoglycemic activities.

     

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