酶法制备牡蛎ACE抑制肽的条件优化  

Enzymatic Preparation of Angiotensin I-Converting Enzyme Inhibitory Peptides Derived from Oyster

作　　者：苑园园[1] 于宏伟[1] 田益玲[1] 苏旭东[1] 马雯[1] 崔同[1] 张伟[1] 檀建新[1] 

机构地区：[1]河北农业大学食品科技学院生物工程系河北省农产品加工工程技术中心,河北保定071000

出　　处：《中国食品学报》2013年第3期115-121,共7页Journal of Chinese Institute Of Food Science and Technology

基　　金：国家海洋局贝类高值化利用技术中试研究与示范项目(200805046);国家自然科学基金资助项目(31071494)

摘　　要：采用检测血管紧张素转换酶(ACE)活性的高效液相色谱法,对自制猪肺ACE酶催化反应条件进行优化,结果表明ACE与底物HGG的最佳反应条件是:底物HGG和ACE的最适反应浓度分别为5mmol/L和12.5mg/mL,反应时间30min。以水解度(DH)为指标,测定时间因素对不同蛋白酶酶解牡蛎蛋白的影响,结果表明:4h时水解基本完全,中性蛋白酶水解度最高,达到89.88%;木瓜蛋白酶水解度最低,为34.51%。探讨了牡蛎蛋白的水解度与ACE抑制活性的关系,结果表明:水解15min后,酶解产物ACE抑制率达较高水平,随时间延长而缓慢增加或减少;其中胃蛋白酶水解牡蛎蛋白酶解物对ACE的抑制率最高,1200U/g底物的添加量、水解4h的条件下,其抑制率可达96.99%,且酶解物可在12h内保持稳定而不被水解。The Angiotensin I-Converting Enzyme （ACE） inhibitory activity was determined by the rapid method of high performance liquid chromatography （HPLC）. Above all, the ACE reaction conditions catalyzed by the ACE isolated from pig lung were optimized. The results indicated that the optimal reaction time of ACE with its substrate HGG was determined at thirty minutes by measuring the amount of hippuric acid liberated from HGG in the ACE reaction. The optimal concentration of the substrate HGG and ACE were 5 mmol/L and 12.5 mg/mL, respectively. In order to study the preparation of antihypertensive peptide derived from oyster, seven enzymes including flavourzyme, papain, pepsin, alcalase, acidic protease, trypsin and neutrase were applied to hydrolyze oyster. The effect of time on the products hydrolyzed by each of proteases was also described on the basis of the degree of hydrolysis （DH）. The results showed that the degree of hydrolysis approximately reached the peak at four hours. Among these seven proteases, neutral protease had the strongest hydrolysis activity and its degree of hydrolysis approach to 89.88%, Contrarily, the degree ,of hydrolysis of papain protease was lowest at 34.51%. The relationship between the inhibitory rate of ACE and the degree of hydrolysis of oyster degraded by the different proteases was discussed. The results showed that the ACE inhibitory rate of the dif- ferent hydrolysates could rise to a high level at the first fifteen minutes, while it revealed a slow increase or even decrease with time prolonging. The hydrolysates by treatment with pepsin possessed the highest ACE inhibitory ratio（96.99%） under the conditions of 1 200 U enzymatic concentration per gram substrate for four hours. Its corresponding products proved to be stable in twelve hours probably due to its low pH range. In addition, the relationship between the inhibition rate of ACE and the degree of hydrolysis was not significant linear correlation.

关 键 词：血管紧张素转换酶 牡蛎 蛋白酶 水解度 抑制率 抑制肽 

分 类 号：TQ464.7[化学工程—制药化工]