灰树花多糖复合酶协同微波辅助提取工艺及抗氧化性研究

Studies on Compound Enzymes Synergistic Microwave-assisted Extraction Process and Antioxidant Activity of Grifola frondosa Polysaccharide

以灰树花干物质为原料,通过响应面Box-Behnken法优化复合酶协同微波辅助提取多糖工艺,采用sevage法去蛋白、001×7大孔树脂脱色,并对DEAE-52纤维素层析柱分离的组分进行抗氧化性研究。响应面试验结果显示,灰树花多糖最佳提取工艺条件为:在微波时间7 min、料水比1∶27(g/mL)、酶配比1∶1(质量比)、酶料比0.02 g/g条件下,灰树花多糖提取量为73.25 mg/g,与预测值的相对偏差为3.17%。采用纯水和0.05 mol/L NaCl对DEAE-52纤维素层析柱进行洗脱得到两种灰树花多糖(Grifola frondosa polysaccharide,GFP)GFP1、GFP2。体外抗氧化试验表明:GFP1、GFP2浓度为6 mg/mL时,还原力达到0.43和0.48;对1,1-二苯基-2-三硝基苯肼(1,1-dipheny1-2-picryl-hydrazyl,DPPH)自由基清除率最高分别为33.54%和50.38%、对羟自由基清除率最高分别为81.59%和90.21%、对超氧阴离子自由基清除率最高分别为79.23%和87.23%,GFP2相较GFP1具有更好的抗氧化能力。

The dry matter of Grifola frondosa was used as raw material,optimization of complex enzymes synergistic microwave-assisted extraction of the polysaccharide by the Box-Behnken method of response surface.The sevage method was used to deproteinize,001×7 macroporous resins were used to decolorize and the components separated by DEAE-52 cellulose chromatography column were analyzed antioxidant.The test results of response surface showed that the optimal extraction conditions for Grifola frondosa polysaccharides were as follows:under the conditions of microwave time of 7 min,solid-water ratio of 1∶27(g/mL),complex enzymes ratio of 1∶1(mass ratio),and enzyme-solid ratio of 0.02 g/g,The extraction of Grifola frondosa polysaccharide was 73.25 mg/g,and the relative error with the expected value was 3.17%.Two kind of polysaccharides GFP1 and GFP2 were eluted by pure water and 0.05 mol/L NaCl on DEAE-52 cellulose chromatography column.The in vitro antioxidant test results showed that the reducing power reached 0.43 and 0.48 when the concentrations of the two polysaccharides were 6 mg/mL.The highest DPPH radical scavenging were 33.54%and 50.38%.The highest hydroxyl radical scavenging were 81.59%and 90.21%.The superoxide anion scavenging rate were 79.23%and 87.23%.The GFP2 had better antioxidant activity than GFP1.