Research Center for Agricultural and Sideline Products Processing, Henan Academy of Agricultural Sciences, Zhengzhou, 450002, P.R. China.
J Food Biochem. 2021 Apr;45(4):e13638. doi: 10.1111/jfbc.13638. Epub 2021 Feb 5.
To prepare and identify ACE-inhibitory peptides originated from sesame seed protein, peptides with strong ACE-inhibitory activities were obtained via the optimization of protease and hydrolysis conditions, and these peptides were purified and identified by membrane separation, gel filtration, and liquid chromatography-mass spectrometry. Results showed that the dual-enzyme comprised alcalase and trypsin with the enzyme activity ratio of 3:7 was suitable to produce ACE-inhibitory peptides. The highest ACE-inhibitory activity of 98.10 ± 0.26% was obtained at the following parameters, pH 8.35, E/S ratio of 6,145 U/g, and hydrolysis time of 4.4 hr. ISGAQPSLR and VVISAPSK ranked the first and second ACE-inhibitory activity among 15 identified ACE-inhibitory peptides. Both peptides influenced ACE via binding with the S1 pocket, S2 pocket, and Zn ion. ISGAQPSLR even impacted the S1' pocket. ISGAQPSLR and VVISAPSK acted as a competitive and noncompetitive inhibitor, respectively. ACE-inhibitory peptides derivated from sesame seed protein have potential applications in functional food. PRACTICAL APPLICATIONS: Although sesame seed protein is proven as the precursor of ACE-inhibitory peptide, preparing ACE-inhibitory peptide from sesame seed protein is still suffering from insufficient information on hydrolysis condition and the peptide sequence. Therefore, the performance of the typical protease on preparing ACE-inhibitory peptide from sesame seed protein has been evaluated, the effect of the amino acid composition of sesame seed protein and cleavage specificity of protease on the generation of ACE-inhibitory peptide has been investigated, hydrolysis conditions have been optimized, the peptide sequence has been identified to illuminate the effect of sesame seed protein fraction on the formation of ACE-inhibitory peptide and discuss the structural characteristics. ACE-inhibitory peptides originating from sesame seed protein could apply in functional food. It is promising for dual-enzyme hydrolysis to utilize in preparation of high-value bioactive peptides.
为了从芝麻蛋白中制备和鉴定具有 ACE 抑制活性的肽，通过优化蛋白酶和水解条件，获得了具有强 ACE 抑制活性的肽，并通过膜分离、凝胶过滤和液相色谱-质谱对这些肽进行了纯化和鉴定。结果表明，复合酶由碱性蛋白酶和胰蛋白酶组成，酶活比为 3:7，适合生产 ACE 抑制肽。在以下参数下，获得了最高的 ACE 抑制活性 98.10±0.26%：pH8.35、E/S 比 6145 U/g 和水解时间 4.4 小时。ISGAQPSLR 和 VVISAPSK 在鉴定的 15 种 ACE 抑制肽中，排名第一和第二。两种肽都通过与 S1 口袋、S2 口袋和 Zn 离子结合来影响 ACE。ISGAQPSLR 甚至影响了 S1'口袋。ISGAQPSLR 和 VVISAPSK 分别作为竞争性和非竞争性抑制剂发挥作用。芝麻蛋白来源的 ACE 抑制肽在功能性食品中有潜在的应用。
虽然芝麻蛋白已被证明是 ACE 抑制肽的前体，但从芝麻蛋白中制备 ACE 抑制肽仍缺乏关于水解条件和肽序列的信息。因此，评估了典型蛋白酶对芝麻蛋白制备 ACE 抑制肽的性能，研究了芝麻蛋白的氨基酸组成和蛋白酶的切割特异性对 ACE 抑制肽生成的影响，优化了水解条件，鉴定了肽序列，以阐明芝麻蛋白组分对 ACE 抑制肽形成的影响，并探讨了结构特征。芝麻蛋白来源的 ACE 抑制肽可应用于功能性食品。双酶水解有望用于制备高附加值的生物活性肽。
