山杏仁蛋白源α-葡萄糖苷酶抑制肽的分离、纯化及鉴定

Separation,Purification,and Identification of α-Glucosidase Inhibitory Peptides from Apricot Kernel Proteins

针对木本油料山杏的蛋白质高值化利用问题,利用蛋白酶Alcalase酶解山杏仁蛋白,以体外α-葡萄糖苷酶抑制能力作为评价指标,筛选高活性α-葡萄糖苷酶抑制肽。山杏仁蛋白酶解液通过超滤、G-25葡聚糖凝胶柱、分子筛以及反相高效液相色谱的分离纯化,最终筛选得到2种高活性的α-葡萄糖苷酶抑制肽,其抑制能力分别为6.6μg/mL和7.0μg/mL。利用质谱和氨基酸测序仪对两种α-葡萄糖苷酶抑制肽的分子质量以及序列结构进行了研究,确认2种山杏仁蛋白源α-葡萄糖苷酶抑制肽分别为色氨酸-丙氨酸(WA)和苏氨酸-色氨酸(TW),其α-葡萄糖苷酶抑制能力的IC_(50)值分别为23.97μmol/L和22.93μmol/L。

Apricot kernel proteins was hydrolyzed by Alcalase to solve the problem of high - value utilization of woody oil apricot kernel, using in vitro α - glucosidase inhibiting ability as the evaluation index to screen high - activ- ity α - glucosidase peptide inhibitors. Two kinds of high - activity α - glucosidase peptide inhibitors were isolated and purified from apricot kernel protein hydrolyzates via ultrafiltration, Sephadex G -25, molecular sieve, and reversed phase high - performance liquid chromatography. The inhibition of the AGA inhibitory peptides was 6.6 μg/mL and 7. 0 μg/mL, respectively. The molecular structure and sequences of the AGA inhibitory peptides were identified through mass spectrometry and amino acid sequencing. The AGA inhibitory peptides from the apricot kernel hydroly- zate were identified as WA and TW. The median inhibitory concentration of WA was 23.97 μ mol/L and that of TW was 22.93 μ mol/L.