Department of Pharmacy and Master Program , Tajen University , Pingtung , Taiwan.
Hospitality and Tourism Research Center , National Kaohsiung University of Hospitality and Tourism , Kaohsiung City 81271 , Taiwan.
J Agric Food Chem. 2019 Jul 17;67(28):7810-7820. doi: 10.1021/acs.jafc.9b01922. Epub 2019 Jul 2.
Antihypertensive peptides were screened from thermolysin hydrolysate of seeds (Jue Ming Zi) using two independent bioassay-guided fractionations, reversed-phase high-performance liquid chromatography (RP-HPLC), and strong cation-exchange (SCX) liquid chromatography coupled with angiotensin I-converting enzyme (ACE) inhibitory assay. The identical peptide in the most active RP-HPLC and SCX fractions was simultaneously de novo sequenced as FHAPWK with high-resolution mass spectrometry. FHAPWK (IC = 16.83 ± 0.90 μM) was further identified as a competitive inhibitor and a true inhibitor on ACE by a Lineweaver-Burk plot and preincubation experiment, respectively. The molecular docking simulation indicated that FHAPWK could interact with several key residues of the ACE active site, which is consistent with the result of the inhibitory kinetics study. Moreover, its antihypertensive effect was demonstrated using the animal model of spontaneously hypertensive rats. It is concluded that FHAPWK is the first reported antihypertensive peptide derived from thermolysin hydrolysate of seeds.
从决明子的胰蛋白酶水解物中筛选出具有降压作用的肽，采用两种独立的生物活性导向分离方法，反相高效液相色谱（RP-HPLC）和强阳离子交换（SCX）液相色谱与血管紧张素 I 转化酶（ACE）抑制测定相结合。在最活跃的 RP-HPLC 和 SCX 馏分中具有相同活性的肽，同时用高分辨率质谱法从头测序为 FHAPWK。FHAPWK（IC = 16.83 ± 0.90 μM）通过 Lineweaver-Burk 图和预孵育实验分别进一步鉴定为 ACE 的竞争性抑制剂和真正抑制剂。分子对接模拟表明，FHAPWK 可以与 ACE 活性部位的几个关键残基相互作用，这与抑制动力学研究的结果一致。此外，还用自发性高血压大鼠动物模型证明了其降压作用。因此，可以得出结论，FHAPWK 是第一个从决明子的胰蛋白酶水解物中分离得到的具有降压作用的肽。
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