Preparation, Purification and Characterization of Antibacterial and ACE Inhibitory Peptides from Head Protein Hydrolysate of Kuruma Shrimp, Marsupenaeus japonicus

Abstract

Kuruma shrimp (Marsupenaeus japonicus) heads, as the main by-product of the seafood processing industry, are rich in underutilized high-quality protein. After papain hydrolysis at 50 °C for 4 h, the protein hydrolysate of shrimp heads was found to show notable antibacterial and angiotensin I-converting enzyme (ACE) inhibitory activities. After purification using two stages of revered-phase high-performance liquid chromatography (RP-HPLC), the antibacterial peptide VTVP and the ACE inhibitory peptide ARL/I were successfully identified from most active fractions by LC–MS/MS. Peptide VTVP was a desirable hydrophobic peptide, with a MIC value in the range from 1.62 to 8.03 mM against all tested pathogens. Peptide ARL/I exhibited potent ACE inhibitory activity, with an IC50 value of 125.58 µM, and was found to be a competitive inhibitor based on the Lineweaver–Burk plot. Moreover, the result of the molecular docking simulation indicated that the interaction binding between ARL/I and ACE was mainly stabilized by hydrogen bonds, as well as forming a coordinate bond with the Zn2+ site. The purified peptides did not show hemolytic activity toward rabbit erythrocytes. To sum up, the bioactive peptides isolated from shrimp heads could be applicable for food or pharmaceutical areas as promising ingredients.

Identification of angiotensin‐converting enzyme inhibitory peptides from peanut meal (Arachis hypogaea Linn) fermented by Lactobacillus pentosus using MALDI‐TOF–MS and LC–MS/MS

“…The analysis of the detected amino acid content ratios revealed that the content of glutamic acid was the highest at 14.94%, followed by leucine and aspartic acid. The results of a related study showed a similar pattern in wheat gluten ACEIP, with gluten content as high as 41.89% (Zhou et al., 2023).…”

“…Following and refining the experimental method proposed by Zhou, Han et al. (2023), 50 mg/mL of post‐purification lyophilized ACEIP sample solutions were prepared. These samples were separated using Sephadex G‐25 column chromatography (HY‐148140, Merck & Co Inc.) at an elution flow rate of 1.0 mL/min and an injection volume of 3 mL.…”

Angiotensin-I-Converting Enzyme (ACE)-Inhibitory Peptides from the Collagens of Monkfish (Lophius litulon) Swim Bladders: Isolation, Characterization, Molecular Docking Analysis and Activity Evaluation

“…The ACEi activity of MH-1b was 76.81 ± 2.76%, which was significantly higher than those of PSC-MH (53.22 ± 2.63%), MH-I (62.75 ± 1.89%), MH-Ia (63.39 ± 3.06%) and MH-Ic (42.77 ± 2.38) (p < 0.05) (Figure 5B). Gel filtration is an efficient technology for separating bioactive substances with different MWs and is widely used to purify ACEi peptides from different hydrolysates of marine proteins, such as Skipjack tuna roes [34], Ulva prolifera [65], Kuruma shrimp [66], Ruditapes philippinarum [67], pearl oyster [68], squilla [39], hybrid groupers [41], etc. Although MH-1b presented the highest ACEi activity, its MW was not the lowest among the three peptide components.…”

“…RP-HPLC is an extremely efficient technology for isolating ACEi peptides according to their RT, and the RT of separated ACEi peptides can be modulated by adjusting the ratio of polar solvent (such as trifluoroacetic acid (TFA), methanol, ethyl alcohol and acetonitrile) in the mobile phase [30,32,64]. Therefore, ACEi peptides have been separated using RP-HPLC from protein hydrolysates of T. bimaculatus [28], R. philippinarum [67], Skipjack tuna roes [31], M. edulis [30], Kuruma shrimp [66], pearl oyster [68], Pyropia pseudolinearis [69], squilla [39], hybrid groupers [41], Arthrospira platensis [70], etc.…”

The Research Progress of Bioactive Peptides Derived from Traditional Natural Products in China

“…In protein hydrolysate separation, SEC and/or membrane separation are often utilized in the initial stage to obtain the desired molecular weight or chain length, followed by further characterization to achieve the desired outcome. For example, Zhou et al used 50 kDa, 10 kDa, 5 kDa, and 1 kDa UF membranes to separate walnut proteolytes into five fractions, which were desalted by a macroporous adsorbent resin and loaded on a Sephadex G-15 chromatographic column to collect the separation obtained at 280 nm walnut ACE inhibitory peptide [57,58]. Notably, SEC generally offers a faster and more efficient separation technique compared to membrane separation, while maintaining a high level of resolution [59].…”