酪蛋白水解物的类蛋白反应修饰及其产物ace抑制活性特征 - 食品科学.pdf

60 2011, Vol. 32, No. 19 食品科学 ※基础研究 ACE 孙 辉，赵新淮 * (乳品科学教育部重点实验室，东北农业大学，黑龙江 哈尔滨 150030) 采用碱性蛋白酶水解酪蛋白，制备水解度为 10.9%、IC50 值为 52.6 μg/mL 的酪蛋白水解物，并利用响应 面法优化碱性蛋白酶催化的类蛋白反应修饰条件。修饰反应时间固定为 6h 时，适宜的条件为酶添加量3.1kU/g pro、 底物质量浓度 50g/100mL 、反应温度25 ℃。制备9 个修饰程度不同的修饰产物，结果显示：修饰产物 ACE 抑制活 性均提高，并且活性最高的修饰产物的 IC50 降低至 14.9 μg/mL 。该修饰产物离心分级后，上清液部分和沉淀部分 的ACE 抑制活性分别低于和高于修饰产物，表明沉淀部分是提高 ACE 抑制活性的主要原因；Tricine-SDS电 泳分析表明，修饰产物及沉淀部分有较大分子质量的肽分子生成；该修饰产物和上清液部分、沉淀部分的进一步 酶水解处理则显示，酶水解会导致它们的 ACE 抑制活性降低，但是仍然高于最初的酪蛋白水解物。 酪蛋白水解物；碱性蛋白酶；类蛋白反应；ACE 抑制活性 Modification of Casein Hydrolysate by Alcalase-catalyzed Plastein Reaction and ACE-inhibitory Activity of Modified Products SUN Hui，ZHAO Xin-huai* (Key Laboratory of Dairy Science, Ministry of Education, Northeast Agricultural University, Harbin 150030, China) Abstract ：Casein hydrolysate with a hydrolysis degree of 10.9% and an ACE-inhibitory IC50 of 52.6 μg/mL was prepared by hydrolyzing casein with alcalase and then modified by alcalase-catalyzed plastein reaction. When the reaction time was fixed at 6 h, the optimal reaction conditions were achieved by response surface methodology to be enzyme dosage of 3.1 kU/g protein, reaction temperature of 25 ℃ and substrate concentration of 50 g/100 mL. Nine modified casein hydrolysates with different modification degrees were prepared. The results indicated that each modified casein hydrolysate had enhanced ACE-inhibitory activity. The IC50 of the modified hydrolysate with the highest activity was 14.9 μg/mL. This modified hydrolysate was centrifuged into two fractions, a supernatant and a precipitation fraction.