Extraction and in vitro simulated gastrointestinal digestion of velvet antler water-soluble protein
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摘要: 以鲜马鹿茸为原料,对比六种蛋白提取方法,依据提取蛋白量及SDS-PAGE电泳图谱确定鹿茸水溶性蛋白有效提取方法;模拟胃肠环境对提取的蛋白进行体外模拟消化研究,测定其体外胃肠消化稳定性及消化过程中游离氨基含量变化。结果表明:组成为100mmol/L Tris、6mol/L盐酸胍、0.02mol/L EDTA-2Na和1%胃蛋白酶抑制剂的提取液提取蛋白量最高,电泳条带多且清晰;蛋白在体外模拟胃、肠环境下消化5min后即迅速被分解,消化60min后即被分解为20ku以下的小分子量蛋白或多肽,消化2.5h后分子量主要分布在7823u及以下范围,在肠和胃肠连续消化过程中游离氨基含量随时间的延长而显著增加,这表明鹿茸水溶性蛋白较易被分解;模拟肠消化2.5h后游离氨基含量增加了一倍,显著大于胃消化中的增加量（p<0.01）,这暗示着鹿茸水溶性蛋白更有利于肠消化。Abstract: The optimal extract method of velvet antler water- soluble protein was determined in this study according to the protein content and SDS-PAGE electrophoresis of six extracts. In vitro simulated gastrointestinal digestion of velvet antler water-soluble protein was investigated in terms of digestibility and free α-amino nitrogen content. Results showed that the extract obtained with the solution composed of 100mmol/L Tris, 6mol/L guanidine hydrochloride, 0.02mol/L EDTA-2Na and 1% Pepstatin A had the largest protein content and clearest electrophoresis bands. The protein was rapidly hydrolyzed in 5min by in vitro simulated gastrointestinal digestion, and then it was broken into small proteins and peptides less than 20 ku in 60 min. The hydrolysate mainly distributed below 7823 u after 2.5h, and the free α-amino nitrogen content was increased significantly during both intestinal and gastrointestinal digestion. The content of free α-amino nitrogen had doubled after simulating intestinal digestion for 2.5h, and was significantly higher than the increment in gastric digestion (p<0.01) . The results indicated that water-soluble protein of velvet antler was easily hydrolyzed and more suitable for intestinal digestion.
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