Antihypertensive Peptides Research Articles

Identification and evaluation of antioxidant activity of bioactive casein-derived peptides during in vitro digestion and transepithelial transport in Caco-2 cells

Dairy products are important sources of bioactive (antibacterial, antithrombotic, antihypertensive, immunomodulatory, and antioxidative) peptides, that need to be released from the target proteins. In the present study, bovine milk casein protein was hydrolyzed by pancreatin and trypsin (4 and 24 h) to evaluate the hydrolyses efficiency (ninhydrin reaction); to elucidate the sequence of casein-derived peptides (SDS-PAGE, LC-ESI-TOF-MS); and to assess the peptides antioxidant activity (DPPH and ORAC), bioactivity in silico, bioaccessibility after in vitro digestion, and bioavailablility by transepithelial transport in Caco-2 cells. The hydrolysis degree increased with the incubation time, with evident degradation of casein-derived proteins into low molecular weight peptides (<5 kDa) after 4 and 24 h incubation. Pancreatin showed higher hydrolysis degree (91%) than trypsin (21%). An increase in antioxidant activity was observed with increasing incubation time, with higher antioxidant effect observed for trypsin derived peptides (47% reduction of free radicals; 32112 μM Trolox equivalents) than pancreatin derived peptides (15% reduction; 2862 μM Trolox equivalents). A total of 69 caseins derived peptides were identified after trypsin (33 peptides) and pancreatin (36 peptides) hydrolysis for 24 h; with 26 peptides released from β-casein, 21 from α-s1-casein, 15 from α-s2-casein, and 7 from k-casein hydrolysis. Some of these peptides were described as antioxidant, anti-inflammatory, and antihypertensive. After in vitro digestion 123 peptides were found in gastric (56 peptides), duodenal (43 peptides) and colonic (24 peptides) fraction; while five peptides were bioavailable by crossing the intestinal barrier, with increased peptides’ abundance after trypsin hydrolysis treatment.

Valorisation of pork by-products to obtain antioxidant and antihypertensive peptides

The porcine liver could be used for the extraction of zinc-protoporphyrin (ZnPP) as a natural red meat pigment. During the autolysis process, porcine liver homogenates was incubated at pH 4.8 and 45 °C under anaerobic conditions to obtain insoluble ZnPP. After incubation, the homogenates were readjusted at pH 4.8, and at pH 7.5 before being centrifuged at 5500 × g for 20 min at 4 °C and the resulting supernatant were compared with the obtained at pH 4.8 at the beginning of the incubation. The molecular weight distributions of the porcine liver fractions at both pHs were very similar, however, eight essential amino acids were more abundant in fractions obtained at pH 4.8. Regarding the ORAC assay, porcine liver protein fraction at pH 4.8 showed the highest antioxidant capacity but antihypertensive inhibition was similar for both pHs. Peptides with strong bioactivity potential from aldehyde dehydrogenase, lactoylglutathione lyase, SEC14-like protein 3 and others were identified. The findings have demonstrated the potential of the porcine liver to extract natural pigments and bioactive peptides.

Restraint of VP1 Protein of Foot and Mouth Disease Virus using Specific Antiviral Peptides: an in Silico Investigation

Foot and mouth diseases are among the important threats in the animal husbandry industry which lead to huge economic losses. In this regard, the current project aimed to inhibit the VP1 protein of foot and mouth disease viruses using specific peptides. For this purpose, a wide range of potential antiviral peptides were collected from the database. Physicochemical properties, hydrophobicity/hydrophilicity, and solubility properties of potential antiviral peptides were investigated using reliable servers. Afterward, the tertiary structures of the selected peptides along with the VP1 protein were modeled by the I-TASSER server. Moreover, interactions between VP1 protein and selected antiviral peptides were investigated using the ClusPro 2.0 server. Finally, the outputs of molecular docking were assessed by LigPlot+ and visualized by PyMol software. The results revealed that Dermaseptin-3, Ginkbilobin, Circulin-F, Maximin1, Cycloviolin-A, Cycloviolin-D, Circulin-C, Cycloviolin-C, and Antihypertensive protein BDS-1 peptides with a hydrophobicity value of &gt; 30 were soluble with positive instability index and positive net charge. Moreover, the results of the molecular docking process demonstrated that Dermaseptin-3 and Ginkbilobin peptides could strongly inhibit the VP1 protein using 10 hydrogen bonds. Therefore, these two peptides, which had the most hydrogen bonds, were introduced as the best anti-foot and mouth disease virus peptides to apply.

Machine annealing-guided navigation of antihypertensive food peptide selectivity between human ACE N- and C-domains in structurally interacting diversity space.

Human angiotensin-converting enzyme (ACE) is a well-established druggable target for the treatment of hypertension (HTN), which contains two structurally homologous but functionally distinct N- and C-domains. Selective inhibition of the C-domain primarily contributes to the antihypertensive efficiency and can be exploited as medicinal agents and functional additives for regulating blood pressure with high safety. In this study, we used a machine annealing (MA) strategy to guide the navigation of antihypertensive peptides (AHPs) in structurally interacting diversity space with the two ACE domains based on their crystal/modeled complex structures and an in-house protein-peptide affinity scoring function, aiming to optimize the peptide selectivity for C-domain over N-domain. The strategy generated a panel of theoretically designed AHP hits with a satisfactory C-over-N (C > N) selectivity profile, from which several hits were found to have a good C > N selectivity, which is roughly comparable with or even better than the BPPb, a natural C > N-selective ACE-inhibitory peptide. Structural analysis and comparison of domain-peptide noncovalent interaction patterns revealed that (i) longer peptides (>4 amino aids) generally exhibit stronger selectivity than shorter peptides (<4 amino aids), (ii) peptide sequence can be divided into two, section I (including peptide C-terminal region) and section II (including peptide middle and N-terminal regions); the former contributes to both peptide affinity (primarily) and selectivity (secondarily), while the latter is almost only responsible for peptide selectivity, and (iii) charged/polar amino acids confer to peptide selectivity relative to hydrophobic/nonpolar amino acids (that confer to peptide affinity).

Cover Feature: Rational Design of a Highly Specific Prolyl Endopeptidase To Activate the Antihypertensive Effect of Peptides (ChemBioChem 8/2023)

Bioactive peptides produced by enzymatic hydrolysis of food-derived proteins could be used for various biomedical applications such as antihypertensive drugs. In the depicted study, a highly specific M. xanthus prolyl endopeptidase mutant was rationally designed by constraint/free molecular dynamics simulations and binding energy calculations towards the application in the hydrolysis of food-derived proteins to produce antihypertensive peptides with a proline C-terminus. More information can be found in the Research Article by J. Z. H. Zhang, L. Zhang, et al.

In-depth peptidomic profile and molecular simulation studies on ACE-inhibitory peptides derived from probiotic fermented milk of different farm animals

Investigations into ACE inhibitory properties of probiotic fermented bovine, camel, goat, and sheep milk were performed and studied for two weeks of refrigerated storage. Results from the degree of proteolysis suggested higher susceptibility of goat milk proteins, followed by sheep and camel milk proteins, to the probiotic-mediated proteolysis. ACE-inhibitory properties displayed continuous decline in ACE-IC50 values for two weeks of refrigerated storage. Overall, goat milk fermented with Pediococcus pentosaceus caused maximum ACE inhibition (IC50: 262.7 µg/mL protein equivalent), followed by camel milk (IC50: 290.9 µg/mL protein equivalent). Studies related to peptide identification and in silico analysis using HPEPDOCK score revealed presence of 11, 13, 9 and 9 peptides in fermented bovine, goat, sheep, and camel milk, respectively, with potent antihypertensive potential. The results obtained suggest that the goat and camel milk proteins demonstrated higher potential for generating antihypertensive peptides via fermentation when compared to bovine and sheep milk.

Bioactivities and production of antihypertensive peptides during fermentation of soy milk by lactic cultures

The objective of this study was to evaluate the effect of pH on fermentative characteristics of selected lactic acid bacteria in soy milk, determining α-galactosidase activity and the utilization pattern of sucrose, raffinose and stachyose during fermentation of soymilk and also to evaluate the angiotensin-converting enzyme (ACE) inhibitory and proteolytic activities. In this study, Lactobacillus helveticus MTCC5463 (V3) reduced maximum pH (pH 4.25) in soy milk (pH 6.5), produced highest viable lactic counts, i.e. 13.32 log CFU/ml in soy milk (pH 8.0) during fermentation. Highest acidity was observed in case of V3 (0.75% lactic acid (LA)) in soy milk (pH 6.5) during fermentation. Overall, lactic acid bacteria performed well in natural soy milk (pH 6.5) as compared to soy milks having pH 7.0 and pH 8.0. V3 also showed highest α-galactosidase activity in comparison to other lactic cultures at pH 6.5. Similarly, Lactobacillus rhamnosus MTCC 5945 (NS4) utilized maximum amount of sucrose and stachyose after 24 h of incubation than other cultures at pH 6.5. NS4 (L. rhamnosus MTCC 5945) culture showed the highest ACE inhibition activity (65.33%) and the highest proteolytic activity (3.264 mg/ml) in soymilk (pH 6.5). The results suggested that the strains (V3 and NS4) could be used for the development of fermented soy milks to reduce complex soy oligosaccharides.

Recent Advances in In Vitro and In Vivo Studies of Antioxidant, ACE-Inhibitory and Anti-Inflammatory Peptides from Legume Protein Hydrolysates.

Consumption of legumes has been shown to enhance health and lower the risk of cardiovascular disease and specific types of cancer. ACE inhibitors, antioxidants, and synthetic anti-inflammatories are widely used today; however, they have several undesirable side effects. Thus, researchers have focused on finding ACE inhibitors, antioxidant, and anti-inflammatory peptides from natural sources, such as legumes. Recently, in vitro and in vivo research has shown the bioactive peptides generated from legume protein hydrolysates, such as antioxidant, anti-hypertensive, anticancer, anti-proliferative, anti-inflammatory, etc., in the context of different disease mitigation. Therefore, this review aims to describe the recent advances in in vitro and in vivo studies of antioxidant, anti-hypertensive and anti-inflammatory peptides isolated from legume-derived protein hydrolysates. The results indicated that antioxidant legumes peptides are characterized by short-chain sequence amino acids and possess anti-hypertensive properties by reducing systolic blood pressure (SBP) in spontaneously hypertensive rats (SHR).

Structural, spectroscopic, molecular docking, ADME, molecular dynamics studies of Val-Trp dipeptide

Hypertension is a significant risk factor for various diseases, especially heart, brain, and kidney diseases. It is known that many peptides have the property of lowering blood pressure and determine as ACE inhibitors. The purpose of this study is to obtain information about the molecular structure of Val-Trp (L-valyl-L-tryptophan), which is one of the antihypertensive peptides, by molecular mechanical, quantum mechanical, and spectroscopic methods (FT-IR and Raman). Also, it is aimed to investigate the interactions of Val-Trp dipeptide with receptors related to hypertension and to determine the pharmacokinetic profile due to the potential of the peptide to be a drug candidate. The peptide structure was optimized by DFT/B3LYP/6-311++G(d,p) basis set, then vibrational wavenumbers, molecular electrostatic potential (MEP), HOMO-LUMO (highest occupied molecular orbital- lowest unoccupied molecular orbital), NBO (natural bond orbital) analyzes were performed. The assignment of fundamental theoretical vibration wavenumbers was carried out with potential energy distribution analysis (PED). After the structural analyzes of the peptide were performed, the interactions of the peptide with Angiotensin-converting enzyme (ACE), Angiotensin II Receptor Type 1 (AT1R) and Renin were investigated by molecular docking study. Then, the molecular dynamic (MD) simulation of the peptide-ACE complex with the best binding affinity in the molecular docking studies was carried out for 50 ns. ADME (absorption, distribution, metabolism, and excretion) analysis of Val-Trp dipeptide was performed. In support of the studies carried out, enlightening information about the feasibility of the antihypertensive drug of Val-Trp dipeptide with the help of the ADME profile was presented to the literature.Communicated by Ramaswamy H. Sarma

Identification of ACE I-Inhibitory Peptides Released by the Hydrolysis of Tub Gurnard (Chelidonichthys lucerna) Skin Proteins and the Impact of Their In Silico Gastrointestinal Digestion.

Tub gurnard is a highly abundant fishery species caught as a discard in the Mediterranean Sea. This work proposes its valorisation through the release of potential antihypertensive peptides and glycosaminoglycans (GAGs) through the controlled hydrolysis of tub gurnard skin proteins. Four proteases (Esperase, Alcalase, Trypsin and Pronase E) were used to obtain potent angiotensin converting enzyme I (ACE)-inhibitory hydrolysates. Peptides and GAGs were separated and evaluated for their antihypertensive potential by fluorometry. The peptide-rich fractions derived from the Esperase and Alcalase hydrolysates showed very low IC50 values (47 and 68 μg/mL, respectively). Only the GAGs from the Trypsin and Esperase hydrolysates were relevant ACE inhibitors (63 and 52% at 1 mg/mL, respectively). The peptide composition of the most potent ACE-inhibitory fractions derived from the Esperase and Alcalase hydrolysates (IC50 values of 33 and 29 μg/mL, respectively) was analysed by RP-LC-ESI-MS/MS. The analysis suggests that the ACE-inhibitory activity is related to the peptide hydrophobicity, as well as to the presence of specific residues at any of the last four C-terminal positions. The in silico gastrointestinal digestion of these fractions yielded small peptides with antihypertensive potential.

Fast Skeletal Muscle Troponin and Tropomyosin as a Dietary Source of Antidiabetic and Antihypertensive Bioactive Peptides: An In Silico Study

Background : The nutraceutical properties of food hydrolysates rely on multiple biochemical interactions involving the modulation of enzymes and cellular receptors. Numerous bioactive peptides released from troponin and tropomyosin digestion have been identified. Their characterization has mostly been performed by hydrolysis catalyzed by proteases unrelated to the human digestive system. Objective: This study aimed to determine the bioactive profile of beef, pork, and chicken meat by analyzing the frequency and pharmacokinetics of biopeptides released from troponin and tropomyosin. Methods: In silico digestion and biopeptide release frequency were studied by three parameters; bioactive fragments release frequency (AE), frequency percentage (W), and mean occurrence (AS), all stated on the BIOPEP-UWM platform. Further on, hydrolysis end-products were screened based on gastrointestinal-absorption probability and pharmacokinetic profiling performed on SwissADME, SwissTargetPrediction, and ADME/Tlab bioinformatics web tools. Statistical analyses were performed using a one-way ANOVA test. Results: Dipeptidyl peptidase-IV (DPP-IV) and angiotensin-converting enzyme (ACE) inhibiting biopeptides exhibited the highest release frequency. Moreover, W and AS parameters showed no significant difference (p&gt;0.05) between the myofibrillar isoforms assessed. Seven biopeptides were classified as highly absorbable and reported optimal drug-likeness compliance. Although biopeptides hold good pharmacokinetic properties, the therapeutic potency of biopeptides showed to be lower than those of DPP-IV and ACE-inhibiting drugs. Conclusions: Troponin and tropomyosin are rich dietary sources of bioactive peptides, mainly DPP-IV and ACE inhibitors. Digestion end-products are mainly dipeptides with optimal pharmacokinetic and drug-like properties, suggesting a potential therapeutic application in hypertensive and hyperglycemic disorders.

Study on the preparation process of quinoa anti-hypertensive peptide and its stability.

Quinoa seeds are a food resource rich in protein, vitamins, minerals, and other functional components such as polyphenols, polysaccharides, and saponins. The seeds have become favored by modern consumers due to being gluten-free and featuring a high protein content. This study focused on the preparation of quinoa peptides by short-time enzymatic-assisted fermentation. Quinoa flour (QF) was mixed with water in a certain ratio before being enzymatically digested with 0.5% amylase and 0.1% lipase for 6 h. Then, 16 bacterial taxa were used for fermentation, respectively. The peptide content in the resulting fermentation broths were determined by the biuret method. The dominant taxon was then identified and the peptide content, amino acid distribution, and molecular weight distribution of the prepared quinoa peptides were analyzed. Further, the temperature, pH, metal ions, organic solvents, ion concentration, and anti-enzyme stability of the quinoa anti-hypertensive peptides of different molecular weights after fermentation with the dominant taxon were investigated. Finally, the inhibitory activity of fermented quinoa peptides on bacteria was studied. The results show that the peptide content of the fermentation broth reached 58.72 ± 1.3% at 40 h of fermentation with Lactobacillus paracasei and the molecular weights of the hydrolyzed quinoa peptides were mainly distributed below 2 kDa by polyacrylamide gel. The Angiotensin Converting Enzyme (ACE) inhibition and peptide retention of the 0-3 kDa quinoa peptides were screened to be high and stable. At the same time, the inhibitory activity of quinoa peptide after fermentation on E. coli was obvious. This study provides a theoretical basis for further research on quinoa peptide and its application in industrial production, and also lays a foundation for the later application of polypeptides in new food and chemical products.

Egg white protein hydrolysate decreased blood pressure via the competing endogenous RNA regulatory networks in female spontaneously hypertensive rats.

Despite numerous studies having reported the effects and mechanisms of antihypertensive peptides including peptides derived from egg white proteins, the role of peptides in a female hypertensive animal model is unknown. On the other hand, the role of epigenetic modulation by peptide treatment has rarely been investigated. This study sought to investigate the effect of egg white protein hydrolysate (EWH) in female spontaneously hypertensive rats (SHRs) as well as to explore the underlying mechanisms from the perspectives of the transcriptome and the profiles of non-coding RNAs. Young (12-14-week-old) female SHRs were orally administered 250 mg per kg body weight (low-dose) or 1000 mg per kg body weight (high-dose) EWH daily for 10 weeks. The blood pressure of the rats was monitored weekly. The mRNA and non-coding RNAs (miRNA, lncRNA, and circRNA) in the aorta were profiled by the high-throughput RNA-seq technique. Differentially expressed (DE) RNAs in the aorta were identified for the construction of the competing endogenous RNA (ceRNA) networks and key molecules were validated by qRT-PCR. The treatment of the high-dose EWH showed a significant effect on reducing blood pressure in female SHRs. Bioinformatic analyses revealed 813, 90, 347 and 869 DE-mRNAs, DE-miRNAs, DE-lncRNAs and DE-circRNAs, respectively. The CNTN5-LncRNA-XR_001835895.1-miR-384-5p was identified as the central network which was validated in the aorta and circulation of female SHRs. The results from this study demonstrated that the treatment with EWH reduced blood pressure via regulating the ceRNA networks in female SHRs, which provided novel insights into the mechanisms of food protein-derived antihypertensive peptides.

Probiotic fermented whey-milk beverages: Effect of different probiotic strains on the physicochemical characteristics, biological activity, and bioactive peptides

The effect of probiotic strains (Lactobacillus acidophilus La-03 (La-03); Lactobacillus acidophilus La-05 (La-05); Bifidobacterium Bb-12 (Bb-12) or Lacticaseibacillus casei-01 (L. casei-01)) on the characteristics of fermented whey-milk beverages during storage (4 °C, 30 days) was evaluated. The products were assessed for biological and antioxidant activities, physicochemical characteristics, and bioactive peptides. Probiotic addition increased α-amylase and α-glucosidase inhibition and antioxidant activities, mainly at 15 days of storage. L. casei-01 showed higher metabolic activity (higher titratable acidity and lower pH values) and the presence of anti-hypertensive peptides, while La-5 and Bb-12 showed higher α-glucosidase inhibition, improvements in the high saturated hypercholesterolemic index, and peptides with ACE-inhibitory, antimicrobial, immunomodulatory, and antioxidant activities. Our findings suggest that probiotic fermented whey-milk beverages may exert antidiabetic and antioxidant properties, being suggested La-5 or Bb-12 as probiotics and 15 days of storage.

Novel antihypertensive peptides from lupin protein hydrolysate: An in-silico identification and molecular docking studies

Application of non-thermal treatment to proteins prior to enzymatic hydrolysis can facilitate the release of novel bioactive peptides (BPs) with unique biological activities. In this study, lupin protein isolate was pre-treated with ultrasound and hydrolysed using alcalase and flavourzyme to produce alcalase hydrolysate (ACT) and flavourzyme hydrolysate(FCT). These hydrolysates were fractionated into 1, 5, and 10 kDa molecular weight fractions using a membrane ultrafiltration technique. The in vitro angiotensin-converting enzyme (ACE) studies revealed that unfractionated ACT (IC50 = 3.21 mg mL−1) and FCT (IC50 = 3.32 mg mL−1) were more active inhibitors of ACE in comparison to their ultrafiltrated fractions with IC50 values ranging from 6.09 to 7.45 mg mL−1. Molecular docking analysis predicted three unique peptides from ACT (AIPPGIPY, SVPGCT, and QGAGG) and FCT (AIPINNPGKL, SGNQGP, and PPGIP) as potential ACE inhibitors. Thus, unique BPs with ACE inhibitory effects might be generated from ultrasonicated lupin protein.

Antidiabetic, angiotensin‐converting enzyme inhibitory and anti‐inflammatory activities of fermented camel milk and characterisation of novel bioactive peptides from lactic‐fermented camel milk with molecular interaction study

This study was to separate and purify antidiabetic and angiotensin‐converting enzyme (ACE) inhibitory peptides from Lactiplantibacillus plantarum KGL3A fermented camel milk. After 48 h of fermentation at 37°C, ɑ‐amylase inhibition, ɑ‐glucosidase inhibition, lipase inhibition and ACE inhibitory activities were 80.94%, 64.45%, 63.93%, and 77.53%, respectively in fermented camel milk. Optimisation of growth condition for the evaluation of maximum peptide production was evaluated by measuring proteolytic activity (O‐phthalaldehyde, OPA method) with different inoculation rates and incubation times and highest proteolytic activity (9.21 mg/mL) was observed after 48 h of fermentation at 2.5% rate of inoculation. The antidiabetic and ACE inhibitory activity of 3 kDa permeate fraction were higher as compared with other fractions. Purification of antidiabetic and ACE inhibitory peptides from fermented camel milks was performed through sodium dodecyl‐sulphate polyacrylamide gel electrophoresis, and 2‐dimensional polyacrylamide gel electrophoresis and maximum number of protein bands were present in between 25 and 10 kDa. The generated peptide sequences were matched with antihypertensive peptide database (AHTPDB) and BIOPEP databases for confirming the antidiabetic and ACE inhibitory activity. Peptides, that is. TDVMPQWW and MMSLVSLLLVGILFPTIQAK were having highest peptide ranker score among the all sequences. Furthermore, anti‐inflammatory activity (TNF‐α, IL‐6 and IL‐1β) of fermented camel milk was evaluated in macrophage cell line RAW 264.7 (Ralph and William's cell line). Furthermore, peptides were predicted to have improved binding affinity against Human Angiotensin converting enzyme (hACE) through molecular docking.

Core-shell lipid-polymeric nanoparticles for enhanced oral bioavailability and antihypertensive efficacy of KY5 peptide

Hypertension is the leading risk factor for death and disability, and hypertensive patients always need long-term oral antihypertensive drugs. Some bioactive peptides that extracted from animals or plants have shown excellent advantages on antihypertension. However, the oral delivery of these peptides is always failure on account of instability and poor absorption in the gastrointestinal tract. Herein, we developed a core-shell lipid-polymeric nanoparticle for oral delivery of a highly efficient antihypertensive peptide KY5 (KY5-CSs). KY5-CSs had a particle size of 216.7 ± 2.5 nm, with a narrow PDI of 0.07 ± 0.01. The zeta potential was −4.1 ± 0.1 mV. It exhibited good stability in 4 °C and possessed a controlled release behavior in gastrointestinal tract. The cellular uptake study proved that the lipid shell imparted unique capability of permeation across the mucus layer and internalization by Caco-2/HT-29 cells. In addition, KY5-CSs enhanced in situ intestinal absorption in SD rats. The pharmacokinetic studies and antihypertensive efficacy showed a superior oral absorption and antihypertensive effect of KY5-CSs than KY5-NPs. In conclusion, the core-shell lipid-polymeric nanoparticles will provide attractive potential for oral delivery of antihypertensive peptides.

Deep learning drives efficient discovery of novel antihypertensive peptides from soybean protein isolate

As a potential and effective substitute for the drugs of antihypertension, the food-derived antihypertensive peptides have arisen great interest in scholars recently. However, the traditional screening methods for antihypertensive peptides are at considerable expense and laborious, which blocks the exploration of available antihypertensive peptides. In our study, we reported the use of a protein-specific deep learning model called ProtBERT to screen for antihypertensive peptides. Compared to other deep learning models, ProrBERT reached the highest the area under the receiver operating characteristic curve (AUC) value of 0.9785. In addition, we used ProtBERT to screen candidate peptides in soybean protein isolate (SPI), followed by molecular docking and in vitro validation, and eventually found that peptides LVPFGW (IC50 = 20.63 μM), VSFPVL (2.57 μM), and VLPF (5.78 μM) demonstrated the good antihypertensive activity. Deep learning such as ProtBERT will be a useful tool for the rapid screening and identification of antihypertensive peptides.

Integrated in silico-in vitro molecular modeling and design of halogenated phenylalanine-containing antihypertensive peptide inhibitors with halogen bonds to target human angiotensin-I-converting enzyme

Angiotensin-I-converting enzyme (ACE) is a key regulator of blood pressure as its critical role in the renin-angiotensin-aldosterone and kallikrein-kinin systems. Although a variety of natural and food peptides have been discovered to possess ACE-inhibitory activity over the past decades, they cannot be directly used as drugs due to their low efficiency and bad bioavailability. In this study, the intermolecular interaction of ACE with a snake venom inhibitor of bradykinin potentiating peptide b (BPPb) was investigated systematically at structural and energetic levels. The C-terminal region of the peptide can tightly pack against the enzyme active site and is primarily responsible for the enzyme inhibition. A tetrapeptide that covers the C-terminal region of BPPb exhibited a satisfactory ACE-inhibitory profile, which was used as start to generate a number of phenylalanine (Phe)-containing antihypertensive peptide inhibitors. The ortho-, meta- and para-positions of peptide Phe−2 phenyl moiety were systematically substituted with halogen atoms. It is revealed that bromine substitution separately at the meta- and para-positions can considerably improve the peptide inhibitory activity by 7.6-fold and 3.7-fold by forming a bifurcated halogen-bonding system with ACE His513 and Val518 residues as well as a binary halogen-bonding system with ACE Glu384 residue, respectively.

Identification of Gastrointestinal Digestion Stable Antihypertensive Fish Peptides from Atlantic Mackerel (Scomber scombrus).

High blood pressure has been recognized as one of the main risk factors of cardiometabolic syndrome by health organizations. Hypertension is medicated by various classes of synthetic drugs; however, adverse effects have repeatedly been reported. Moreover, natural alternatives such as fish peptides have been effective in the treatment and prevention of hypertension. The aim of our study was to fractionate and identify antihypertensive peptides. Fractions were produced using different techniques of solid-phase extraction (SPE), pressure-driven ultrafiltration (UF), and electro04dialysis with UF membrane. According to our results, the hydrophobic fraction of SPE (IC50 5 μg) was the most potent anti-angiotensin converting enzyme (ACE) product. Findings of the study suggest that the separation technique plays an important role in the isolation efficiency of antihypertensive biopeptides. Importantly, the hydrophobic fraction's activity was retained through a static model of an in vitro gastrointestinal digestion system. In conclusion, polarity regardless of charge and size was the most important factor for anti-ACE activity of an Atlantic mackerel biopeptide. In addition, the presence of leucine at either of the extremities (C- or N-terminal) and/or leucine-rich motifs could well explain the hypotensive effect of our active fraction.