水-酶联合法制备灵芝多糖及其抗氧化活性研究

Preparation of Ganoderma lucidum polysaccharides by water-enzyme combination method and their antioxidant activity

中文关键词:  灵芝  多糖  水-酶联合法  抗氧化活性

英文关键词:Ganoderma lucidum  polysaccharide  water-enzyme combination method  antioxidant activity

中文摘要:

      目的 结合热水浸提法和酶提法的优点, 采用水-酶联合法制备灵芝多糖(Ganoderma lucidum polysaccharide-water-enzyme, GLP-W-E)并研究其结构特征和抗氧化活性。方法 以提取得率为指标, 采用响应面实验优化热水浸提法和水-酶联合法工艺条件, 并以此工艺条件制备热水浸提多糖(Ganoderma lucidum polysaccharide, GLP)和GLP-W-E, 比较了两种多糖的红外光谱、单糖组成及抗氧化活性。结果 热水浸提法最佳条件为: 液料比30:1 (V:m)、温度97℃、时间2.5 h; 酶法最佳条件为: 酶底比1600:1 (U/g)、温度47℃、时间1.8 h。最优条件下, 水-酶联合法提取的GLP-W-E得率为1.66%, 比GLP(两次水提的提取得率为1.45%)提高了14.48%。傅里叶变换红外光谱法(Fourier transform infrared spectroscopy, FT-IR)表明, GLP和GLP-W-E均为吡喃环酸性多糖。高效阴离子交换色谱-脉冲安培检测法(high performance anion exchange chromatography-pulsed amperometric detection, HPAEC-PAD)结果表明, GLP和GLP-W-E单糖组成相似(均有Fuc、GlcN、Gal、Glc、Xyl、Man、GluA 7种单糖), 但其摩尔百分比有明显不同。二者的抗氧化活性也有明显不同, GLP-W-E与GLP相比有明显提高, 尤其是GLP-W-E的2,2’-联氮-二(3-乙基-苯并噻唑啉-6-磺酸)二铵盐[2,2’-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) ammonium salt, ABTS]阳离子自由基清除率比GLP提高了52.09%。结论 水-酶联合法制备的灵芝多糖提取得率更高、活性更强。

英文摘要:

      Objective To prepare Ganoderma lucidum polysactical-water-enzyme by water-enzyme combined with the advantages of hot water extraction and enzyme extraction, and study its structure and antioxidant activity. Methods With the extraction rate as the index, Ganoderma lucidum polysaccharide (GLP) and GLP-W-E were prepared by optimizing the process conditions of hot water extraction method and water-enzyme combination method using response surface test, and the infrared spectra, monosaccharide composition and antioxidant activity of the 2 kinds of crude polysaccharides were compared. Results The optimal conditions of water extraction were as follows: Liquid-solid ratio 30:1 (V:m), temperature 97℃, time 2.5 h; the optimum conditions for enzymatic method were: Enzyme base ratio 1600:1 (U/g), temperature 47℃, time 1.8 h. Under the optimal conditions, the yield of GLP-W-E extracted by water-enzyme was 1.66%, which was 14.48% higher than that of GLP extracted by twice water extracts (1.45%). Fourier transform infrared spectroscopy (FT-IR) showed that GLP and GLP-W-E were both pyranocyclic polysaccharide. The results of high performance anion exchange chromatography-pulsed amperometric detection (HPAEC-PAD) showed that GLP and GLP-W-E monosaccharides were similar in composition (they were all composed of 7 kinds of monosaccharides: Fuc, GlcN, Gal, Glc, Xyl, Man and GluA), but their molar ratios were significantly different. The antioxidant activity of GLP-W-E was significantly higher than that of GLP, especially the 2,2’-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) ammonium salt (ABTS) cationic free radical clearance rate of GLP-W-E was 52.09% higher than that of GLP. Conclusion The polysaccharides prepared by water-enzyme combination method has higher extraction rate and stronger activity.