两种双酶切大豆分离蛋白技术路线的比较-食品科学.pdfVIP

※基础研究 食品科学 2008, Vol. 29, No. 05 65 两种双酶切大豆分离蛋白技术路线的比较 宋春丽，迟玉杰 * (大豆生物学教育部重点实验室，东北农业大学食品学院，黑龙江 哈尔滨 150030) 摘 要：本研究以碱性丝氨酸蛋白酶和蛋白酶Ⅱ作为水解酶对大豆分离蛋白进行两步定向酶切，以制备具有改善 肝昏迷生物活性的高F值寡肽。本研究设定两种定向酶切的技术路线，即在酶解过程中维持碱性丝氨酸蛋白酶最适 pH的时间分别设定为3h(技术路线Ⅰ)和30min(技术路线Ⅱ)，而蛋白酶Ⅱ分别在酸性及中性条件下继续酶解大豆分 离蛋白水解液。结果表明，技术路线Ⅰ、Ⅱ蛋白质回收率分别为86.5%、87.8%；酸溶性氮得率分别为83.2%、 86.4%；灰分含量分别为11.1%、7.0%；技术路线Ⅰ所得肽的相对分子质量分布范围为：分子量大于5000 的肽占 肽总量的20.6%，分子量在3000左右的肽占总量的 3.3%，分子量在500～600之间的肽占肽总量的61.3%；技术路 线Ⅱ所得的产物组分相对分子质量分布范围为：分子量在1000以下的占71.3%，且分子量主要在700～900之间， 分子量在2000 左右的占5.7%，分子量在5000 以上的占 12.8%。研究结果表明，技术路线Ⅱ的酶切效果优于技术 路线Ⅰ的。 关键词：碱性丝氨酸蛋白酶；蛋白酶Ⅱ；大豆分离蛋白；寡肽 Comparison of Two Technologies for Soybean Protein Isolate Two-Enzymatic Hydrolysis SONG Chun-li，CHI Yu-jie* (Key Laboratory of Soybean Biology, Ministry of Education, College of Food Science, Northeast Agricultural University, Harbin 150030, China) Abstract ：The procedure to obtain oligopeptide with soybean protein isolate by sequential treatment with two proteinases was described. The experiment were designed : technique one (T1) kept optimum pH of proteinaseⅠfor 3h by adding NaOH, technique two (T2) kept optimum pH of proteinaseⅠonly at first 30 minutes and hydrolysate are immediately hydrolyzed by proteinaseⅡon acid and neutral condition respectively. The results showed that protein recovery rate are 86.5 %, 87.8 % and acid soluble nitrogen rate are 83.2 %, 86.4 % and ash contents are 11.1% and 7.0% respectively. The molecular weight distribution of peptides obtained by T1 above 5000, about 3000 and between 500 and 600 are 20.6%，3.3% and 61.3% respectively; The molecular weight distribution of peptides obtained by T2 above 5000, about 2000 and between 700 and