ACE-inhibitory Activity Research Articles

Prediction and Validation of Proline-containing Tripeptides with Angiotensin I-converting Enzyme Inhibitory Activity Using Machine Learning Models

Background: Numerous inhibitory peptides against angiotensin I-converting enzyme, a target for hypertension treatment, have been found in previous studies. Recently, machine learning screening has been employed to predict unidentified inhibitory peptides using a database of known inhibitory peptides and descriptor data from docking simulations. Objective: The aim of this study is to focus on angiotensin I-converting enzyme inhibitory tripeptides containing proline, to predict novel inhibitory peptides using the machine learning algorithm PyCaret based on their IC50 and descriptors from docking simulations, and to validate the screening method by machine learning by comparing the results with in vitro inhibitory activity studies. Methods: IC50 of known inhibitory peptides were collected from an online database, and descriptor data were summarized by docking simulations. Candidate inhibitory peptides were predicted from these data using the PyCaret. Candidate tripeptides were synthesized by solid-phase synthesis and their inhibitory activity was measured in vitro. Results: Seven novel tripeptides were found from the peptides predicted to have high inhibitory activity by machine learning, and these peptides were synthesized and evaluated for inhibitory activity in vitro. As a result, the proline-containing tripeptide MPA showed high inhibitory activity, with an IC50 value of 8.6 µM. Conclusion: In this study, we identified a proline-containing tripeptide with high ACE inhibitory activity among the candidates predicted by machine learning. This finding indicates that the method of predicting by machine learning is promising for future inhibitory peptide screening efforts.

Microencapsulation of Hibiscus sabdariffa extract in taro starch: Determination of optimal conditions of bioactive compound retention using response surface methodology

Hibiscus sabdariffa (HS) extract contains a large amount of bioactive compounds with antioxidant, anticarcinogenic, and antihypertensive effects. The extraction of total phenols, flavonoids, and anthocyanins is more efficient at boiling temperature (~91 °C) than that performed at room temperature (~27 °C). In this study, a central composite rotatable design (CCRD) was used to optimize the inlet temperature conditions of a spray dryer and the concentration of taro starch solids to obtain microcapsules with the highest retention of bioactive compounds in hibiscus extracts. Optimized microcapsules (OM) were obtained at an inlet temperature of 118 °C and solid concentration of 26.5 %. Low moisture content (4.54 %) and water activity (0.2) and high content of total phenols (3374.91 mg GAE/100 g), flavonoids (372.81 mg QE/100 g), monomeric anthocyanins (36.74 mg C-3-GE/100 g), and angiotensin-converting enzyme inhibitory activity (80.01 %), were determined in OM using response surface methodology. OM showed a range of spherical agglomerate sizes (10––32 μm). These results indicate that CCRD is a good tool for establishing the optimal conditions for solid concentration and drying temperature to maximize the protection of bioactive compounds using taro starch as the wall material.

Potential blood pressure regulatory effect of low molecular weight α-chymotrypsin extract and its peptides from Stichopus japonicus: Peptide-ACE interaction study via in silico molecular docking

Hypertension is a significant risk factor for cardiovascular disease and heart damage. Stichopus japonicus (S. japonicus) is a red sea cucumber with antioxidant and anti-hypertensive activities. However, its anti-hypertensive mechanism remains unclear. In this study, we evaluated the inhibitory effects of angiotensin-converting enzyme (ACE) of α-chymotrypsin-assisted hydrolysate of S. japonicus (α-chy) and its ultrafiltrate fractions (>10 kDa [α-chy-I], 5–10 kDa [α-chy-II], and <5 kDa [α-chy-III]). Low molecular α-chy-III exerted excellent ACE inhibitory and anti-hypertensive activities in spontaneously hypertensive rats (SHR). Moreover, α-chy-III significantly reduced the systolic and diastolic blood pressure by modulating the serum angiotensin and ACE levels. Histological analysis revealed that α-chy-III markedly protected the aorta, heart, and kidney tissues of SHR from hypertension-induced tissue damage.Taken together, our findings highlight the potential of α-chy-III as a functional food ingredient for the treatment of hypertension and its comorbidities.

Antihypertensive peptides from hydrolyzed proteins of Pleurotus spp.: production, in vitro digestion and identification

This study aimed to investigate in vitro antihypertensive property of protein hydrolysates from Pleurotus spp. mushrooms obtained through enzymatic hydrolysis. FlavourzymeTM 500L, AlcalaseTM 2.4L and NeutraseTM 0.8L were used alone and in combination using an experimental design of mixtures. Antihypertensive activity was determined by angiotensin-converting enzyme (ACE) inhibition and the simulated in vitro digestion was performed according to the INFOGEST protocol. Results showed that most of the protein hydrolysates obtained displayed higher ACE inhibitory activity than non-hydrolyzed protein, ranging from 15.76% and 50.87% inhibition. The highest ACE inhibition and the lowest TCA soluble protein content (52.09%) were detected for the protein hydrolysates obtained by using the binary mixture of FlavourzymeTM 500L and AlcalaseTM 2.4L in equal proportions. Hydrolysis kinetics showed no significant difference in ACE inhibitory activity between 20 and 120minutes of enzymatic reaction. The fraction with molecular weight between 3 and 5kDa obtained after ultrafiltration showed the most contribution for ACE inhibitory activity (47% inhibition). After simulated digestion, the hydrolysates maintained a significant ACE inhibition capacity, indicating the resistance of the peptides to the action of gastrointestinal enzymes. Through proteomic analysis, 26 peptides were identified and the sequences LPILP, IPLLP, PLLPQ and VIQYDPPQ were considered potentially antihypertensive.

Enzymatic hydrolysis of duck blood protein produces stable bioactive peptides: Pilot-scale production, identification, and stability during gastrointestinal and plasma digestion

This study addresses the valorization of duck blood, an underutilized protein-rich by-product from the poultry industry, into bioactive protein hydrolysates with antioxidant and ACE inhibitory properties. Raw and heat-treated duck blood were compared as substrates for enzymatic hydrolysis using Neutrase and Papain. Gel electrophoresis revealed that heat treatment reduced fibrinogen content, while FTIR analysis showed that heat treatment modified the protein structure, increasing β-sheet content from 21.13 % to 34.96 %. Heat-treated duck blood hydrolyzed by Neutrase exhibited superior hydrolysis (9.53 %) and protein recovery (60.28 %) compared to raw blood. Pilot-scale production (1000 L) enhanced hydrolysate yield and maintained bioactive properties. LC-MS/MS analysis identified five novel bioactive peptides derived from the hydrolysate's 4-h simulated gastrointestinal (GI) digestion, with WMHVR demonstrating the highest antioxidant and ACE inhibitory activities. Molecular docking simulations revealed that WMHVR competitively inhibits ACE by binding to the S1, S2, and S′ pockets through van der Waals and hydrogen bonding interactions. The GI-hydrolysate and identified peptides maintained bioactivity during simulated GI and blood plasma digestion, with ACE inhibition increasing over time. This research transforms food industry waste into functional protein hydrolysates, offering applications in nutraceuticals and functional foods while promoting sustainable practices through waste protein valorization.

DeepBP: Ensemble deep learning strategy for bioactive peptide prediction

BackgroundBioactive peptides are important bioactive molecules composed of short-chain amino acids that play various crucial roles in the body, such as regulating physiological processes and promoting immune responses and antibacterial effects. Due to their significance, bioactive peptides have broad application potential in drug development, food science, and biotechnology. Among them, understanding their biological mechanisms will contribute to new ideas for drug discovery and disease treatment.ResultsThis study employs generative adversarial capsule networks (CapsuleGAN), gated recurrent units (GRU), and convolutional neural networks (CNN) as base classifiers to achieve ensemble learning through voting methods, which not only obtains high-precision prediction results on the angiotensin-converting enzyme (ACE) inhibitory peptides dataset and the anticancer peptides (ACP) dataset but also demonstrates effective model performance. For this method, we first utilized the protein language model—evolutionary scale modeling (ESM-2)—to extract relevant features for the ACE inhibitory peptides and ACP datasets. Following feature extraction, we trained three deep learning models—CapsuleGAN, GRU, and CNN—while continuously adjusting the model parameters throughout the training process. Finally, during the voting stage, different weights were assigned to the models based on their prediction accuracy, allowing full utilization of the model's performance. Experimental results show that on the ACE inhibitory peptide dataset, the balanced accuracy is 0.926, the Matthews correlation coefficient (MCC) is 0.831, and the area under the curve is 0.966; on the ACP dataset, the accuracy (ACC) is 0.779, and the MCC is 0.558. The experimental results on both datasets are superior to existing methods, demonstrating the effectiveness of the experimental approach.ConclusionIn this study, CapsuleGAN, GRU, and CNN were successfully employed as base classifiers to implement ensemble learning, which not only achieved good results in the prediction of two datasets but also surpassed existing methods. The ability to predict peptides with strong ACE inhibitory activity and ACPs more accurately and quickly is significant, and this work provides valuable insights for predicting other functional peptides. The source code and dataset for this experiment are publicly available at https://github.com/Zhou-Jianren/bioactive-peptides.

Untargeted metabolomics profiling for revealing water-soluble bioactive components and biological activities in edible bird’s nest

Edible bird’s nest (EBN) is a functional food renowned for its numerous health benefits. While its nutritional and therapeutic value is well-documented, the metabolites contributing to the bioactivities of EBN remain poorly understood. This study aimed to identify the metabolites present in EBN subjected to different treatments, including double-boiled EBN (EBNdb), EBN hydrolysate (EBNhydro), EBN fermented with Lactobacillus helveticus (EBNLH), Latilactobacillus curvatus (EBNLC), and Latilactobacillus sakei (EBNLS) using liquid chromatography-mass spectrometry (LC-MS) and correlate the identified bioactive metabolites with the bioactivities of EBN. It was found that the fermented EBNs exhibited the highest number of metabolites, with 76 tentatively identified, followed by EBNhydro (45) and EBNdb (37). Citric acid (1.97–4.48 g/kg) was present in all treated EBN samples, while L(+)-lactic acid (3.03–8.07 g/kg) and adipic acid (2.33–3.18 g/kg) were only found in fermented EBNs. Among the treated EBN samples, EBNLC demonstrated the significantly highest (p ≤ 0.05) antioxidative (22.34 ± 0.41 % 1,1-diphenyl-2-picrylhydrazil radical scavenging activity), antihypertensive (5.46 ± 0.26 % angiotensin-converting enzyme inhibitory activity), and antihyperglycemic activities (6.48 ± 0.34 % α-amylase inhibitory activity). A total of 18 metabolites, including citric acid, 4-sphingenin, N-acetylcitrulline, 4-aminophenol, L(+)-lactic acid, 2-oxoadipate, sildenafil, formylglycinamidin-RP, 11β,17α,21-α-5β-pregnane-3,20-dione, 2-ketobutyric acid, homoserine, benzaldehyde, 1-pyrroline4-hydroxy-2-carboxylate, nortriptyline, 1-methylguanine, 3-hydroxy-trimethyllysine, 3-phenylpropionate, and reserphine were predicted as bioactive metabolites using the partial least squares discriminant analysis (PLS-DA). This study provides valuable insights into the metabolites present in EBN and serves as fundamental data for future investigations into the bioactive compounds responsible for its specific health benefits, potentially leading to the development of enhanced EBN-based functional foods.

Feta cheese: A pool of Non-Starter Lactic Acid Bacteria (NSLAB) with anti-hypertensive potential

In the present work, we examined the angiotensin-I-converting enzyme-inhibitory (ACE-I) activity of the water soluble extracts of four long-ripened Feta cheese samples. High activity (>87 %) was determined for all of them. Additionally, we fingerprinted the non-starter lactic acid bacteria (NSLAB) isolated from the cheese samples. All NSLAB isolates were screened for their proteolytic activity and the most proteolytic ones were further evaluated for their ACE-I activity as well as ability to produce γ-aminobutyric acid (GABA). The NSLAB isolates comprised 130 lactobacilli and 24 enterococci, with the dominant species being Levilactobacillus brevis, Lactiplantibacillus plantarum, Lacticaseibacillus paracasei and Enterococcus faecium. Among them, 40 selected isolates exhibited high proteolytic activity, with 13 lactobacilli and two enterococci exhibiting strong ACE-I activity (>50 %), and six lactobacilli producing GABA. Thus, the hypertension risk associated with over-consumption of Feta cheese because of its high sodium content may be compensated by the anti-hypertensive traits of NSLAB microbiota members, which are active during the long cheese ripening time.

Antioxidant, Antihypertensive, and Anti-Inflammatory Activities of Long-Term Ripened Cheddar Cheese Water-Soluble Extract.

During cheese ripening, the proteins in the cheese are broken down, and various functional peptides are produced. This research aimed to investigate the changes in peptides and their physiological activities during the long-term maturation of Cheddar cheese. Young (YC), medium (MC), and 3-year-aged extra-sharp (EC) Cheddar cheeses were extracted with water, and bioactive peptides were identified using ultra performance liquid chromatography-high resolution mass spectrometer. Peptides reported to have antioxidant, angiotensin-converting enzyme (ACE)-inhibitory, and anti-inflammatory effects were identified and evaluated in the extracts. MC and EC showed stronger antioxidant activity than YC. The 2,2'-azino-bis-(3-ethylbenzothiazoline-6-sulfonic) acid inhibition rates of MC and EC were similar, but EC exhibited a higher 2,2-diphenyl-1-picrylhydrazyl inhibition rate. The antihypertensive effect was found to increase owing to the appearance of peptides with ACE-inhibitory activity in MC and EC. Quantitative real time polymerase chain reaction and immunoblot were run to evaluate the anti-inflammatory effects. YC did not show anti-inflammatory activity, but MC and EC were shown to effectively inhibit inflammatory mRNA expression. The immunoblot results showed that EC did not inhibit IκBα phosphorylation, but had an inhibitory effect at the mRNA expression level. Overall, the peptides contained in aged Cheddar cheese were shown to have excellent antioxidant, anti-inflammatory, and antihypertensive activities, and long-term ripening appeared to have a positive effect on these activities. This is presumed to have affected not only the already identified peptides but also unknown peptides; therefore, it is expected that the discovery of bioactive peptides will be possible through additional research.

Predicting variable-length ACE inhibitory peptides based on graph convolutional network

Traditional molecular descriptors have contributed to the prediction of angiotensin I-converting enzyme (ACE) inhibitory peptides, but they often fall short in capturing the complex structure of the molecule. To address these limitations, this study introduces molecular graphs as an advanced method for peptide characterization. Peptides containing 2–10 amino acids were represented using molecular graphs, and a graph convolutional network (GCN) model was constructed to predict variable-length peptides. This model was compared with machine learning (ML) models based on molecular descriptors, including Random Forest (RF), Support Vector Machine (SVM), and k-Nearest Neighbor (kNN), under the same benchmark. Notably, the GCN model outperformed the other models with an accuracy of 0.78, effectively identifying ACE inhibitory potential. Furthermore, the GCN model also demonstrated superior performance, exceeding existing methods with an accuracy rate of over 98 % on an independent test set. To validate our predictions, we synthesized peptides VAPE and AQQKEP with high predicted probabilities, and their IC50 values of 2.25 ± 0.11 and 3.75 ± 0.17 μM, respectively, indicating potent ACE inhibitory activity. The developed GCN model presents a powerful tool for the rapid screening and identification of ACE inhibitory peptides, offering promising opportunities for developing antihypertensive components in functional foods.

Effects of fig and pineapple powder on metabolite compounds and health-promoting properties in fermented goat meat sausage.

The objective of this research was to identify the metabolite compounds and health-promoting properties of goat meat fermented sausages containing different amounts of fig and pineapple powder (0.1%, 0.25%, and 0.5%), which are presented by F1, F2, F3 and P1, P2, P3, separately. Fermented sausages were manufactured from the lean meat of female goats. The samples extracted from the goat meat fermented sausages were evaluated for their metabolite compounds, antioxidants, and angiotensin-converting enzyme inhibitory activity. The results showed that the diphenyl picrylhydrazyl radical scavenging activity, azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) radical scavenging activity, and angiotensin-converting enzyme inhibitory activity were the highest in F3 (p<0.05), whereas the hydroxyl radical (°OH) scavenging capacity was the highest in P3 (p<0.05). In addition, F3 and F2 showed the highest superoxide anion radical scavenging activity (p<0.05), whereas F3 and P3 showed the highest copper chelating activity (p<0.05). Based on the metabolite compounds, hydrophobic amino acids, aromatic amino acids, and bitter amino acids were abundant in F3. Both F3 and P3 contained high concentrations of umami-producing compounds. The incorporation of 0.5% fig powder into goat meat fermented sausage showed improved biological activities based on metabolite compounds.

Identification of Structure-Linked Activity on Bioactive Peptides from Sea Cucumber (Stichopus japonicus): A Compressive In Silico/In Vitro Study.

A sea cucumber (Stichopus japonicus) is an invertebrate rich in high-quality protein peptides that inhabits the coastal seas around East Asian countries. Such bioactive peptides can be utilized in targeted disease therapies and practical applications in the nutraceutical industry. Bioactive peptides were isolated from Stichopus japonicus through ultrafiltration and Sephadex G-10 size exclusion chromatography. The low-molecular-weight fraction (ACSH-III) showed the highest hydroxyl radical scavenging and angiotensin-converting enzyme (ACE) inhibitory activities. Subsequent purification of ACSH-III resulted in four fractions, of which ACSH-III-F3 and ACSH-III-F4 exhibited significant bioactivity. Peptides identified in these fractions, including Phenylalanine-Proline-Threonine-Tyrosine (FPTY) and Tyrosine-Proline-Serine-Tyrosine-Proline-Serine (YPSYPS), were characterized using high-performance liquid chromatography (HPLC) and quadrupole time-of-flight mass spectrometry (QTOF-MS). FPTY demonstrated the most potent antioxidant and antihypertensive activities among these peptides, with IC50 values of 0.11 ± 0.01 mg/mL for hydroxyl radicals and 0.03 ± 0.01 mg/mL for ACE inhibition. Docking simulations revealed strong binding affinities of these peptides to the active site of the ACE, with FPTY displaying interactions similar to those of the synthetic inhibitor lisinopril. These findings suggest that the identified peptides, particularly FPTY, have potential applications as natural antioxidants and functional foods.

Influence of the Blends of Sheep's and Goat's Milk on the Functional Properties and Volatile Compounds of Lighvan Cheese During Ripening: A Comparative Study.

In this study, the influence of different ratios of sheep's to goat's milk (100:0, 75:25, 50:50, 25:75, and 0:100) on the content of functional metabolites and volatiles in Lighvan cheese throughout ripening was investigated. The samples were ripened under brine for 90 days and assessed in terms of variations in chemical composition, proteolysis, angiotensin-converting enzyme, α-amylase, and α-glucosidase inhibitory activities, antioxidant power, total free amino acids, gamma amino butyric acid, free fatty acids, and volatile compounds. Results showed that by increasing the proportion of goat's milk, the samples exhibited higher antioxidant effect and inhibition of the angiotensin-converting enzyme during ripening. After 90 days, an increase in α-glucosidase and a slight decrease in α-amylase inhibitory activities were observed in Lighvan cheese samples that were produced with 50%, 75% and 100% of goat's milk. In contrast, higher contents of sheep's milk led to a significant increase in the concentration of esters, terpenes and alcohols and a slight increase in gamma-aminobutyric acid. The highest concentration of saturated and unsaturated fatty acids was observed in the cheese prepared with 100% raw sheep's milk after 90 days of ripening. The results revealed the possibility of making a functional white-brined cheese by monitoring the type and ratio of the used milk and ripening time.

Research on the Synergistic Inhibition of Angiotensin-Converting Enzyme (ACE) by the Gastrointestinal Digestion Products of the ACE Inhibitory Peptide FPPDVA.

To gain a deeper understanding of the ACE inhibition effect, the inhibitory effect of ACE-inhibiting peptide (ACEIP) FPPDVA's digestive products on ACE was further investigated. Two novel peptides, PD (IC50 = 161.1 ± 1.10 μM) and DV (IC50 = 66.51 ± 0.99 μM) were identified in the digestive products of FPPDVA using LC-MS/MS. The Peptide Mix (FPPDVA, PD, and DV) exhibited a remarkable synergistic effect on ACE inhibition by significantly enhancing it by up to 508% compared to the individual peptides alone. Furthermore, theoretical simulations suggest that the Peptide Mix synergistically inhibits ACE activity by forming more stable complexes with the active site of ACE, facilitated by an increased number of hydrogen bonds. Additionally, Lineweaver-Burk plot analysis and spectroscopic studies further verified the presence of these stable complexes. ITC results show that the combination of Peptides Mix and ACE is a spontaneous exothermic process driven by entropy. The study showed that FPPDVA has a stronger inhibitory effect on ACE after digestion, making it suitable as an antihypertensive peptide in functional foods.

ACE inhibitory effect and saltiness-enhancing properties of chicken-derived umami peptides: Digestive stability, inhibition kinetics, multiple ligand docking and central composite design

Angiotensin-I-converting enzyme (ACE) inhibitory activity and saltiness-enhancing properties of chicken-derived umami peptides were investigated. DGGRYY and NEFGYSNR were screened and the IC50 values were 28.71 μM and 283.24 μM, indicating their potential as novel ACE inhibitors. DGGRYY and NEFGYSNR have good pH and thermal stability. After gastrointestinal digestion, the ACE-inhibitory activity of DGGRYY retained about 53 %, whereas NEFGYSNR retained about 57 %. The inhibition pattern of both peptides was determined to be uncompetitive, consisting with the result of multiple ligand docking that the binding sites were outside the ACE active pocket. Trp59, Tyr62, Asp121, Arg124, and Ser516 were the key binding sites that contributed to the total binding energy. In addition, saltiness and palatability models were established according to sensory analysis and central composite design. In 0.1 % ∼ 0.3 % NaCl solutions, the addition of DGGRYY and NEFGYSNR could enhance the salty intensity and compensate for the palatability loss caused by salt reduction.

The Antioxidant Effects of Trypsin-Hydrolysate Derived from Abalone Viscera and Fishery By-Products, and the Angiotensin-I Converting Enzyme (ACE) Inhibitory Activity of Its Purified Bioactive Peptides.

Abalone is a rich source of nutrition, the viscera of which are discarded as by-product during processing. This study explored the biological activities of peptides derived from abalone viscera (AV). Trypsin-hydrolysate of AV (TAV) was purified into three fractions using a Sephadex G-10 column. Nine bioactive peptides (VAR, NYER, LGPY, VTPGLQY, QFPVGR, LGEW, QLQFPVGR, LDW, and NLGEW) derived from TAV-F2 were sequenced. LGPY, VTPGLQY, LGEW, LDW, and NLGEW exhibited antioxidant properties, with IC50 values of 0.213, 0.297, 0.289, 0.363, and 0.303 mg/mL, respectively. In vitro analysis determined that the peptides VAR, NYER, VTPGLQY, QFPVGR, LGEW, QLQFPVGR, and NLGEW inhibited ACE, with IC50 values of 0.104, 0.107, 0.023, 0.023, 0.165, 0.004, and 0.146 mg/mL, respectively. The binding interactions of ACE-bioactive peptide complexes were investigated using docking analysis with the ZDCOK server. VTPGLQT interacted with HIS513 and TYR523, and QLQFPVGR interacted with HIS353, ALA354, GLU384, HIS513, and TYR523, contributing to the inhibition of ACE activity. They also interacted with amino acids that contribute to stability by binding to zinc ions. QFPVGR may form complexes with ACE surface sites, suggesting indirect inhibition. These results indicate that AV is a potential source of bioactive peptides with dual antioxidant and anti-hypertensive dual effects.

Anti-hypertensive, anti-inflammatory, glycemic and satiety impact of cocoa flavoured soy-based powder drink mix enriched with bioactive hydrolysates from nests of Aerodramus fuciphagus

Abstract Objectives The edible bird’s nest hydrolysates (EBNH) are renowned for their health benefits. The study aimed to determine their contributory functional effect as an ingredient in the cocoa-flavoured soy-based powder drink mix (PDM). The product’s glycemic and satiety impact were also studied. Materials and Methods The product was enriched with a reasonably low amount of spray-dried EBNH. The product’s functional effect with respect to in vitro angiotensin converting enzyme (ACE) inhibitory activity and human red blood cells membrane stabilization (RBCMS) activity were evaluated, using its water extract (90 - 95 oC). The glycemic and satiety impact were evaluated using 12 and 30 subjects, respectively, who consumed 250 mL of the product, containing 25 g of available carbohydrates. Results The enriched PDM’s ACE inhibitory activity and RBCMS activity were significantly higher (p&amp;lt;0.05) than those that were devoid of EBNH. A 240 mL (12.5% w/v) of PDM (containing by weight 3.0% EBNH, 75.44% soybean powder, 20% cocoa powder, 0.5% xanthan gum, 1.0% silicon dioxide and 0.06% sucralose) provides about 3.7 kcal/g energy. Its 250 mL meal (consistency index of 14.17 Pa.sn) is high in protein and dietary fiber. The product’s glycemic index (GI) and glycemic load (GL) are low, 24 and 6.0, respectively, raising a normal level of 2 hours postprandial blood glucose (&amp;lt;6 mmol/L) and satisfies hunger for about 2.5 hours. Conclusion The EBNH, as an ingredient, enhances the PDM’s anti-hypertensive and anti-inflammatory properties. The overall findings suggest that the formulated PDM is a wellness food product capable of promoting good human health, including for dieters and individuals with diabetes.

The profile of plant bioactive compounds in Xinjiang, China unifloral safflower honey and their inhibitory on ACE, and determination of the mechanism of inhibition

Investigating the plant bioactive compounds in unifloral safflower honey from Xinjiang, China, is helpful for verifying its botanical origin and evaluating its functional properties. Therefore, this study quantifying hydroxysafflor yellow A (HSYA), anhydroxysafflor yellow B (ASYB), quercetin, kaempferol, rutin, myricetin, hyperoside, gallic acid, quinic acid, and ferulic acid in safflower honey by high-performance liquid chromatography; HSYA and ASYB accounted for 16.12–28.83% and 13.63–33.09%, respectively, of the quantified plant bioactive compounds and were considered characteristic markers for authentication. These compounds also exhibit angiotensin converting enzyme (ACE) inhibitory activity; the IC50 ranges from 13.33 to 132.45 μmol L−1, and the inhibition constant (Ki) is between 17.13 and 121.34 μmol L−1. HSYA (IC50 = 20.24 μmol L−1, Ki = 19.63 μmol L−1) and ASYB (IC50 = 19.33 μmol L−1, Ki = 17.13 μmol L−1) exhibited the strongest inhibitory effects. Molecular simulation studies have shown that these bioactive compounds bind key residues in the active pocket of ACE through hydrogen bonding and hydrophobic interactions, leading to ACE inhibition. Overall, HSYA and ASYB could be considered bioactive chemical markers for verifying the botanical origin of Xinjiang, China, unifloral safflower honey, and safflower honey may be a potential functional food by inhibiting ACE.

Integrative approach to assessing bioactivity from hempseed protein isolate extracted and dehydrated by different methods: Synergising in silico prediction and in vitro validation

This study demonstrated a comprehensive workflow combining in silico screening and prediction with in vitro validation to investigate the bioactivity of hempseed protein isolate (HPI) extracted and dehydrated using different methods. By adopting an in silico approach, 13 major proteins of HPI were hydrolysed by 20 selected enzymes, leading to the prediction of 20 potential bioactivities. With papain hydrolysis, dipeptidyl peptidase-IV (DPP4) and angiotensin-converting enzyme (ACE) inhibitory activities emerged as having the highest potential. In vitro experiments confirmed these predictions, with DPP4 and ACE inhibitory activities displaying IC50 values of 0.32–0.42 mg/mL and 6.8–9.17 μg/mL, respectively. A strong correlation (r2 = 0.96) was observed between in vitro protein inhibitory results and in silico predicted data. This study demonstrated an effective integrative approach for predicting bioactive peptides in food protein, providing valuable guidance on its processing to create value-added products.

Protein-added kefir: biochemical changes in in vitro digestion stages.

While yogurt is the leading fermented milk product, kefir is at the top of the beverage scale. Milk proteins, on the other hand, show specific functions that positively affect healthy nutrition due to the bioactive components, that they provide the necessary amino acids for growth and development. In our study, kefir, a functional product enriched with whey proteins, casein and skimmed milk powder, which are the natural components of milk, was produced. Added-protein kefir samples were applied the in vitro digestion protocol, static method. In order to observe different protein behaviors, samples were taken pre-digestion, at 120th minute and at 240th minute of digestion protocol. ACE and Antioxidant capacity determination analyzes were carried out. While ACE inhibition values were in the range of 78.63-90.30% pre-digestion, they changed in the range of 86.97-96.38% after gastrointestinal digestion. It was determined that the ACE inhibition values of the control sample remained at the lowest level at all stages of digestion and that the difference between all of samples was significant (P < 0.05). Antioxidant activity value ranging from 0.3615-0.5512 meq Ascorbic acid/μg before digestion was determined as 1.3796-1.9313 meq Ascorbic acid/μg after gastrointestinal digestion (P < 0.05). Kefir samples containing whey protein stand out with their high potential in terms of both antioxidant activity capacity and ACE inhibition activity at all stages of digestion. Considering their therapeutic effects in fermented products, it is thought that whey proteins among milk proteins will be important alternative sources to enrich the protein content in kefir production. © 2024 Society of Chemical Industry.