Instituto de agroquimica y Tecnologia de Alimentos (CSIC), P.O. Box 73, 46100 Burjassot, Valencia, Spain.
J Agric Food Chem. 2010 Mar 10;58(5):2895-901. doi: 10.1021/jf904204n.
The main purpose of this work was to study the generation of Angiotensin I-converting enzyme inhibitory (ACEI) peptides after gastrointestinal digestion of pork meat by the action of pepsin and pancreatin at simulated gut conditions. The hydrolysate was further subjected to reverse phase chromatography in order to separate the fractions with ACEI activity. Using MALDI-TOF/TOF mass spectrometry, 12 peptides were identified in these fractions. It is worth highlighting the novel peptides ER, KLP, and RPR with IC(50) values of 667 microM, 500 microM, and 382 microM, respectively. Results obtained by MALDI-TOF/TOF mass spectrometry were complemented by a second approach consisting of the analysis of the hydrolysate directly by nanoLC-ESI-MS/MS followed by a study of the obtained sequences and comparison with known ACEI peptide sequences. By using these two approaches, a total of 22 peptides were selected for its synthesis and further in vitro assay of ACEI activity. The strongest ACE inhibition was observed for peptide KAPVA (IC(50) = 46.56 microM) followed by the sequence PTPVP (IC(50) = 256.41 microM). Sequence similarity searches revealed that these two peptides derive from muscle titin, constituting the first identified ACEI peptides coming from this protein. This is also the first time that ACEI sequences MYPGIA and VIPEL have been reported. Other identified and synthesized sequences showed less ACEI activity. The obtained results evidence the potential of pork meat proteins as a source of antihypertensive peptides after gastrointestinal digestion.
本工作的主要目的是研究胃蛋白酶和胰蛋白酶在模拟胃肠道条件下作用于猪肉后生成血管紧张素转化酶抑制（ACEI）肽。将所得水解物进一步进行反相色谱分离，以分离具有 ACEI 活性的级分。使用 MALDI-TOF/TOF 质谱，在这些级分中鉴定出 12 种肽。值得强调的是具有 IC50 值分别为 667 μM、500 μM 和 382 μM 的新型肽 ER、KLP 和 RPR。MALDI-TOF/TOF 质谱获得的结果通过另一种方法得到补充，该方法包括直接通过纳升液相色谱-电喷雾-质谱/质谱（nanoLC-ESI-MS/MS）分析水解物，然后研究获得的序列并与已知的 ACEI 肽序列进行比较。使用这两种方法，共选择了 22 种肽进行合成，并进一步进行 ACEI 体外活性测定。观察到最强的 ACE 抑制作用的肽是 KAPVA（IC50=46.56 μM），其次是 PTPVP（IC50=256.41 μM）。序列相似性搜索表明，这两种肽来源于肌肉原肌球蛋白，构成了从该蛋白中首次鉴定出的 ACEI 肽。这也是首次报道 ACEI 序列 MYPGIA 和 VIPEL。鉴定和合成的其他序列显示出较低的 ACEI 活性。所得结果表明，猪肉蛋白质在胃肠道消化后具有作为抗高血压肽的潜在来源。
