大豆粕碱性蛋白酶水解肽及ACE抑制活性的研究（.doc

PAGE 2 PAGE 7 大豆粕碱性蛋白酶水解肽及ACE抑制活性的研究作者简介：温雪琴（1989—），女，（汉），硕士研究生，研究方向：微生物应用及发酵工程。 通信作者：冯欣（1981—），女，（汉），实验师，硕士研究生，微生物应用及发酵工程。 温雪琴1，刘屾1，黄弢1，赵嘉希2，冯欣1*，孙建华1 （1.天津市动植物抗性重点实验室，天津师范大学生命科学学院，天津 300387；2.天津科技大学，天津 300222 ) 摘 要：豆粕是大豆经浸提脱脂后得到的副产品，含有大量的大豆蛋白，其水解物对血管紧张素转移酶（ACE）具有明显的抑制作用和降压效果。结果表明：以食品级豆粕为原料，经碱性蛋白酶水解后得到的ACE抑制肽在pH8.0，温度50 ℃，豆粕底物浓度40 g/L的条件下，酶解3 h得到的产物ACE抑制活性最高，为64.21 ％，对应水解度14.86 ％；进一步对ACE抑制肽进行高效液相色谱（HPLC??以及超高压液相色谱/四极杆-飞行时间串联质谱联用仪(LC-MS)的分析结果显示：活性物质的主要组分为三肽化合物，分子量为359，分子式为C15H30N6O4，由异亮氨酸、精氨酸和丙氨酸构成；其HPLC检测条件为色谱柱：Zorbax300SB-C18，5 μm，4.6×150 mm；检测波长：228 nm，流动相：50 %甲醇-超纯水，各含0.1 %三氟乙酸；流速：0.7 mL/min；柱温：26 ℃；进样量：10 μL。 关键词：豆粕；碱性蛋白酶；ACE抑制肽；高效液相色谱法HPLC； ACE inhibitory Activity on Hydrolytic Peptides from Soybean Meal hydrolyzed by Alkaline Protease WEN Xue-qin1，LIU Shen1，HUANG Tao1，ZHAO Jia-xi2，FENG Xin1*，SUN Jian-hua1 (1.Tianjin Key Laboratory of Animals and Plants Resistance，College of Life Sciences，Tianjin Normal University，Tianjin，300387，PR China；2.Tianjin University of Science and Technology，Tianjin 300222，China) Abstract：Soybean meal, as a byproduct of soybean oil manufacture, contains mainly of protein. Peptides derived from the protein had been proven to be angiotensin-converting enzyme (ACE) inhibitory and antihypertensive effect. Food-grade soybean meal was hydrolyzed by alcalase for the preparation of ACE inhibitory peptides. The optimum enzymolytic conditions for the preparation were at pH8.0, 55 ℃ for 3 h and a substrate concentration of40 g/L. The ACE inhibitory activity and the degree of hydrolysis (DH) were separately high up to 64.21％ and 14.86 %. Subsequent detection of high performance liquid chromatographic (HPLC) and High pressure liquid chromatography / quadrupole - time of flight tandem mass spectrometry (LC-MS) suggested that the active fraction might be tripeptide with molecular weight of 359 and the molecular formula of C15H30N6O4 which constitute of Isoleucine, Arginine and Alanine. Its HPLC conditions were determined as follows: colu