The quality changes, dynamic changes in microbial composition, and diversity changes in large yellow croaker (Larimichthys crocea) during 4 °C refrigeration were studied using 16S rDNA high-throughput sequencing technology, and the total viable count (TVC), total volatile basic nitrogen (TVB-N), and thiobarbituric acid-reactive substances (TBARS) were determined. The results revealed a consistent increase in TVC, TVB-N, and TBARS levels over time. On the 9th day, TVC reached 7.43 lg/(CFU/g), while on the 15th day, TVB-N exceeded the upper limit for acceptable quality, reaching 42.56 mg/100 g. Based on the 16S rDNA sequencing results, we categorized the storage period into three phases: early storage (0th and 3rd days), middle storage (6th day), and late storage (9th, 12th, and 15th days). As the storage time increased, both the species richness and diversity exhibited a declining trend. The dominant genus identified among the spoilage bacteria in refrigerated large yellow croaker was Pseudomonas, accounting for a high relative abundance of 82.33%. A comparison was carried out of the spoilage-causing ability of three strains of Pseudomonas screened and isolated from the fish at the end of storage, and they were ranked as follows, from strongest to weakest: P. fluorescen, P. lundensis, and P. psychrophila. This study will provide a theoretical basis for extending the shelf life of large yellow croaker.

Squid ink melanin can be efficiently extracted from the byproduct ink sac generated during squid processing. As a natural food colorant, it possesses inherent antioxidant properties and the capability to adsorb heavy metals. This study aims to investigate the solubility of water-soluble squid ink melanin (WSSM) obtained from the ink sac, as well as its stability under various conditions including temperature, pH, salt, sugar, potassium sorbate, metal ions, sodium benzoate, sodium sulfite (reducing agent), and hydrogen peroxide (oxidizing agent). Moreover, it explores the scavenging effects of WSSM on free radicals and cadmium ions. The findings suggest that WSSM’s stability is insignificantly affected by high temperature, sucrose, and salt. However, acidity, sodium benzoate, potassium sorbate, sodium sulfite (Na₂SO₃), and hydrogen peroxide (H₂O₂) significantly influence its stability. Most metal ions do not impact the stability of WSSM, except for Fe²⁺, Fe³⁺, Al³⁺, and Cu²⁺, which result in the precipitation of WSSM. Additionally, WSSM exhibits remarkable antioxidant activity with IC₅₀ values of 0.91, 0.56, and 0.52 mg/mL for scavenging superoxide anion radicals (O²⁻·), hydroxyl radicals (·OH), and DPPH radicals, respectively. It also demonstrates the ability to adsorb the heavy metal Cd²⁺, with the adsorption rate gradually increasing with a higher temperature and larger amounts of WSSM added. Infrared spectroscopy analysis reveals the weakening of characteristic peaks (-COOH and -OH) during the process of Cd²⁺ adsorption by WSSM, while SEM confirms surface roughening and structural damage after Cd²⁺ adsorption. This study provides valuable insights for the utilization of squid melanin products as natural antioxidants and heavy metal adsorbents in the food industry.

Candidate peptides with novel angiotensin-I-converting enzyme (ACE) inhibitor activity were obtained from hydrolysates of Gracilariopsis lemaneiformis by virtual screening method. Our results showed that G. lemaneiformis peptides (GLP) could significantly lower blood pressure in spontaneously hypertensive rats (SHR). At least 101 peptide sequences of GLP were identified by LC-MS/MS analysis and subjected to virtual screening. A total of 20 peptides with the highest docking score were selected and chemically synthesized in order to verify their ACE-inhibitory activities. Among them, SFYYGK, RLVPVPY, and YIGNNPAKG showed good effects with IC₅₀ values of 6.45 ± 0.22, 9.18 ± 0.42, and 11.23 ± 0.23 µmoL/L, respectively. Molecular docking studies revealed that three peptides interacted with the active center of ACE by hydrogen bonding, hydrophobic interactions, and electrostatic forces. These peptides could form stable complexes with ACE. Furthermore, SFYYGK, RLVPVPY, and YIGNNPAKG significantly reduced systolic blood pressure (SBP) in SHR. YIGNNPAKG exhibited the highest antihypertensive effect, with the largest decrease in SBP (approximately 23 mmHg). In conclusion, SFYYGK, RLVPVPY, and YIGNNPAKG can function as potent therapeutic candidates for hypertension treatment.

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 (ΔG_total) 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 physicochemical properties of semi-dried Takifugu obscurus fillets in cold air drying (CAD), hot air drying (HAD), and cold and hot air combined drying (CHACD) were analyzed based on pH, water state, lipid oxidation, protein degradation, and microstructure, using a texture analyzer, low-field nuclear magnetic resonance, thiobarbituric acid, frozen sections, sodium dodecyl sulfate polyacrylamide gel electrophoresis, and differential scanning calorimetry. Water binding to the samples was enhanced by all three drying methods, and the immobilized water content of CHACD was between that of HAD and CAD. The pH of the semi-dried fillets was improved by CHACD. When compared to HAD and CAD, CHACD improved the springiness and chewiness of the fillets, especially cold air drying for 90 min (CAD-90), with values of 0.97 and 59.79 g, respectively. The muscle fibers were arranged compactly and clearly in CAD-90, having higher muscle toughness. CHACD reduced the drying time and degree of lipid oxidation compared to HAD and CAD. CAD better preserved protein composition, whereas HAD and CHACD promoted actin production; CHACD had a higher protein denaturation temperature (74.08–74.57 °C). CHACD results in better physicochemical properties than HAD or CAD, including shortened drying time, reduced lipid oxidation, enhanced protein stability, and denser tissue structure. These results provide a theoretical basis for selecting the appropriate drying method for T. obscurus in industrial applications.

An affinity chromatography filler of CNBr-activated Sepharose 4B-immobilized ACE was used to purify ACE-inhibitory peptides from Takifugu flavidus protein hydrolysate (<1 kDa). Twenty-four peptides with an average local confidence score (ALC) ≥ 80% from bounded components (eluted by 1 M NaCl) were identified by LC-MS/MS. Among them, a novel peptide, TLRFALHGME, with ACE-inhibitory activity (IC₅₀ = 93.5 µmol·L⁻¹) was selected. Molecular docking revealed that TLRFALHGME may interact with the active site of ACE through H-bond, hydrophobic, and electrostatic interactions. The total binding energy (ΔGbinding) of TLRFALHGME was estimated to be −82.7382 kJ·mol⁻¹ by MD simulations, indicating the favorable binding of peptides with ACE. Furthermore, the binding affinity of TLRFALHGME to ACE was determined by surface plasmon resonance (SPR) with a Kd of 80.9 µmol, indicating that there was a direct molecular interaction between them. TLRFALHGME has great potential for the treatment of hypertension.