摘要
菌菇来源可溶性β-葡聚糖具有良好的生物活性,为开发利用蛹虫草中的β-葡聚糖(CMBG),本研究以蛹虫草子实体为原料,采用蛋白酶酶解、大孔树脂吸附、DEAE离子交换柱层析进行纯化,以脱色率及β-葡聚糖保留率进行加权评分,摸索建立最佳纯化工艺条件.对获得的β-葡聚糖进行结构分析,并研究其体外降血糖活性.结果显示:大孔树脂HPD-750具有最佳的吸附效果,其吸附最优条件为:上样流速5mL/min、上样浓度4 mg/mL、上样体积400 mL、树脂质量40 g.利用DEAE-FF纤维素交换柱分离出两个β-葡聚糖组分CMBG-1和CMBG-2,相对分子量分别为13.57×104Da和14.36×104 Da;两者均由Gal、Glc、Man三种单糖组成,其摩尔比分别为1.28∶5.27∶1.01和0.71∶4.43∶0.80.通过刚果红分析得出CMBG-1具有三螺旋结构,而CMBG-2不具备三螺旋结构,二者对α-淀粉酶及α-葡萄糖苷酶抑制率都与样品的浓度呈正相关,其对α-淀粉酶抑制的IC50分别为2.68和4.32 mg/mL,对α-葡萄糖苷酶抑制的IC50分别为2.37和3.01 mg/mL,拥有三螺旋结构的CMBG-1的降血糖活性明显高于不具备三螺旋结构的CMBG-2,在功能食品开发方面具有良好的前景.
Abstract
Soluble β-glucan from mushroom source had good biological activity,in order to develop and utilize β-glucan(CMBG)in Cordyceps chrysanthemi,this study used Cordyceps chrysanthemi substrate as raw material,and was purified by proteolytic enzyme digestion,adsorption of macroporous resin,and chromatography of DEAE ion-exchange columns,and the decolorization rate and β-glucan retention rate were weighted and scored to feel out the establishment of the optimal purification process conditions.The obtained β-glucan was structurally analyzed and its in vitro hypoglycemic activity was investigated.The results showed that the macroporous resin HPD-750 had the best adsorption effect,and the optimal conditions for adsorption were as follows:up-sample flow rate of 5 mL/min,up-sample concentration of 4 mg/mL,up-sample volume of 400 mL,and mass of the resin of 40 g.Two β-glucan fractions,CMBG-1 and CMBG-2,with relative molecular weights of 13.57×104 Da and 14.36× 104 Da,respectively,were isolated using a DEAE-FF cellulose exchange column.Both consisted of three monosaccharides,Gal,Glc,and Man,with molar ratios of 1.28∶5.27∶1.01 and 0.71∶4.43∶0.80,respectively.The Congo red analysis showed that CMBG-1 had a triple-helix structure,while CMBG-2 had no triple-helix structure,and the inhibition rates of both of them on α-amylase and α-glucosidase were positively correlated with the concentrations of the samples,and the IC50 of their inhibition on α-amylase were 2.68 and 4.32 mg/mL,and those on α-glucosidase were 2.37 and 3.01 mg/mL,respectively.The hypoglycemic activity of CMBG-1,which possessed a triple-helix structure,was significantly higher than that of CMBG-2,which did not possess a triple-helix structure,and it had good prospects for the development of functional foods.
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