Zhu Qiaonan, Xue Jiawen, Wang Peng, Wang Xianbo, Zhang Jiaojiao, Fang Xuezhi, He Zhiping, Wu Fenghua
College of Food and Health, Zhejiang Agriculture and Forestry University, Hangzhou 311300, China.
Zhejiang Feixiangyuan Food Co., Ltd., Lishui 323400, China.
Foods. 2023 Jan 21;12(3):501. doi: 10.3390/foods12030501.
The camellia seed cake proteins (CP) used in this study were individually hydrolyzed with neutral protease, alkaline protease, papain, and trypsin. The results showed that the hydrolysate had the highest ACE inhibitory activity at 67.36 ± 0.80% after four hours of neutral protease hydrolysis. Val-Val-Val-Pro-Gln-Asn (VVVPQN) was then obtained through ultrafiltration, Sephadex G-25 gel chromatography separation, LC-MS/MS analysis, and in silico screening. VVVPQN had ACE inhibitory activity with an IC value of 0.13 mg/mL (198.66 μmol/L), and it inhibited ACE in a non-competitive manner. The molecular docking indicated that VVVPQN can combine with ACE to form eight hydrogen bonds. The results of the stability study showed that VVVPQN maintained high ACE-inhibitory activity in weakly acidic and neutral environments and that heat treatment (20-80 °C) and Na, Mg, as well as Fe metal ions had little effect on the activity of VVVPQN. Moreover, it remained relatively stable after in vitro simulated gastrointestinal digestion. These results revealed that VVVPQN identified in camellia seed cake has the potential to be applied in functional food or antihypertensive drugs.
本研究中使用的油茶籽饼蛋白（CP）分别用中性蛋白酶、碱性蛋白酶、木瓜蛋白酶和胰蛋白酶进行水解。结果表明，经中性蛋白酶水解4小时后，水解产物的ACE抑制活性最高，为67.36±0.80%。然后通过超滤、Sephadex G-25凝胶色谱分离、LC-MS/MS分析和计算机筛选获得了Val-Val-Val-Pro-Gln-Asn（VVVPQN）。VVVPQN具有ACE抑制活性，IC值为0.13 mg/mL（198.66 μmol/L），并且以非竞争性方式抑制ACE。分子对接表明，VVVPQN可与ACE结合形成八个氢键。稳定性研究结果表明，VVVPQN在弱酸性和中性环境中保持较高的ACE抑制活性，热处理（20-80°C）以及Na、Mg和Fe金属离子对VVVPQN的活性影响很小。此外，它在体外模拟胃肠道消化后仍相对稳定。这些结果表明，油茶籽饼中鉴定出的VVVPQN有潜力应用于功能性食品或抗高血压药物。
