Effects of four kinds of proteases on α-glucosidase inhibitory activity of polysaccharide from Inonotus obliquus
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摘要: 为了探究桦褐孔菌高温水提粗多糖（High temperature water-extracted polysaccharides,HIOP）中结合蛋白质组成键型与其α-葡萄糖苷酶抑制活性的关系,用4种不同的蛋白酶对HIOP进行酶解,测定蛋白酶水解后对其分子量组成及α-葡萄糖苷酶抑制活性的影响。结果显示,与原HIOP在10μg/m L时的α-葡萄糖苷酶抑制率83.72%相比,经中性蛋白酶、碱性蛋白酶、胃蛋白酶和胰蛋白酶酶解处理后的HIOP,α-葡萄糖苷酶抑制率最低分别为53%、65%、6.5%和7.1%,其中胃蛋白酶和胰蛋白酶酶解处理显著降低了HIOP的α-葡萄糖苷酶抑制活性,表明HIOP中结合蛋白在HIOP的α-葡萄糖苷酶抑制活性中有一定作用。结合四种蛋白酶的作用效果与作用键型推测,HIOP中结合蛋白的活性中心可能含有芳香族氨基酸、酸性氨基酸、赖氨酸或精氨酸,破坏此类肽键,α-葡萄糖苷酶抑制活性明显降低,而四种蛋白酶酶解均未使HIOP分子量发生较大改变,说明四种蛋白酶酶解仅影响了HIOP与α-葡萄糖苷酶活性中心结合的部位。
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关键词:
- 桦褐孔菌粗多糖 /
- 蛋白酶 /
- α-葡萄糖苷酶抑制活性
Abstract: The relationship between α-glucosidase inhibitory activity and the composition bond type of binding protein of high temperature water-extracted polysaccharides ( HIOP) that extracted from Inonotus obliquus was studied.HIOP was enzymatically digested with four different proteases, and the effect of protease hydrolysis on its molecular weight composition andα-glucosidase inhibitory activity was determined. The inhibitory rate of α-glucosidase by original HIOP was 83.72% at10 μg/m L, while the lowest α-glucosidase inhibition rate of HIOP that hydrolyzed by neutral protease, alkaline protease, pepsin, and trypsin were 53%, 65%, 6.5% and 7.1%, respectively. The binding protein in HIOP had a role in theα-glucosidase inhibitory activity of HIOP.In addition, in consideration of the effects of these four kinds of enzymes, it could be speculated that the active sites of the binding proteins in HIOP contain aromatic amino acids, acidic amino acids, lysine or arginine.If we destroy such peptide bonds, the α-glucosidase inhibitory activity of HIOP could be significantly reduced.But, the four proteases did not change the molecular weight of HIOP, indicating that the four proteases only affected the binding site of HIOP to α-glucosidase activity center. -
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