Targeted Affinity Purification and Mechanism of Action of Angiotensin-Converting Enzyme (ACE) Inhibitory Peptides from Sea Cucumber Gonads.

Protein hydrolysates from sea cucumber (Apostichopus japonicus) gonads are rich in active materials with remarkable angiotensin-converting enzyme (ACE) inhibitory activity. Alcalase was used to hydrolyze sea cucumber gonads, and the hydrolysate was separated by the ultrafiltration membrane to produce a low-molecular-weight peptide component (less than 3 kDa) with good ACE inhibitory activity. The peptide component (less than 3 kDa) was isolated and purified using a combination method of ACE gel affinity chromatography and reverse high-performance liquid chromatography. The purified fractions were identified by liquid chromatography-tandem mass spectrometry (LC-MS/MS), and the resulting products were filtered using structure-based virtual screening (SBVS) to obtain 20 peptides. Of those, three noncompetitive inhibitory peptides (DDQIHIF with an IC50 value of 333.5 µmol·L-1, HDWWKER with an IC50 value of 583.6 µmol·L-1, and THDWWKER with an IC50 value of 1291.8 µmol·L-1) were further investigated based on their favorable pharmacochemical properties and ACE inhibitory activity. Molecular docking studies indicated that the three peptides were entirely enclosed within the ACE protein cavity, improving the overall stability of the complex through interaction forces with the ACE active site. The total free binding energies (ΔGtotal) for DDQIHIF, HDWWKER, and THDWWKER were -21.9 Kcal·mol-1, -71.6 Kcal·mol-1, and -69.1 Kcal·mol-1, respectively. Furthermore, a short-term assay of antihypertensive activity in spontaneously hypertensive rats (SHRs) revealed that HDWWKER could significantly decrease the systolic blood pressure (SBP) of SHRs after intravenous administration. The results showed that based on the better antihypertensive activity of the peptide in SHRs, the feasibility of targeted affinity purification and computer-aided drug discovery (CADD) for the efficient screening and preparation of ACE inhibitory peptide was verified, which provided a new idea of modern drug development method for clinical use.

The Effect of Ultrasound on the Rehydration Characteristics of Semi-Dried Salted Apostichopus japonicus.

To effectively shorten the rehydration time of Apostichopus japonicus and reduce the nutrient loss during the rehydration process, an ultrasound-assisted rehydration method was adopted to rehydrate semi-dry salted A. japonicus in this study. The effects of different ultrasonic powers, temperatures, and times on the rehydration characteristics, textural characteristics, and sensory quality of the semi-dry salted A. japonicus were studied. Box-Behnken response surface analysis was used to study the influence of the interactions among the three factors on the rehydration ratio of the semi-dry salted A. japonicus, and a quadratic multinomic regression model was established to predict the optimal rehydration ratio. The results showed that ultrasound could change the structure of semi-dry salted A. japonicus and form a spatial network structure, thereby improving its water absorption capacity and reducing rehydration time. The optimal rehydration effect could be obtained when the ultrasonic power was 400 W, the ultrasonic temperature was 50 °C, and the ultrasonic time was 83 min. Ultrasonic power, ultrasonic time, and ultrasonic temperature influenced the rehydration ratio of the semi-dry salted A. japonicus. Under the optimal rehydration conditions in this study, the rehydration ratio of semi-dry salted A. japonicus obtained by the test was 2.103, which was consistent with the value predicted by the Box-Behnken response surface method.