ACE-inhibitory Activity Research Articles

High accuracy prediction of dipeptide angiotensin-converting enzyme (ACE) inhibitory activity by improving the credibility of the 3D-quantitative structure-activity relationship (3D-QSAR) model database and investigating inhibition mechanism

The 3D-QSAR model is one of the most effective techniques for predicting the peptide bioactivity based on their structure properties. The accuracy of model prediction is highly dependent on the bio-activity of the peptides in its dataset. However, the same peptide used for constructing the database was reported to have obviously different activities in different published articles, which may significantly decrease the prediction accuracy. Based on this, we chose ACE inhibitory (ACE-I) dipeptides that can be absorbed directly into the human small intestine as research object. Ten dipeptides were randomly selected from the literatures for molecular docking with ACE and synthesized to test their activity. Ultimately, a linear equation was developed between bio-activities and docking scores, which was used to revalidate the reliable ACE-I activities of all reported dipeptides for constructing a credible dataset of 3D-QSAR model. The prediction accuracy was significantly enhanced and the model we established had a high Q2 = 0.840 and R2 = 0.998. Furthermore, dipeptide CW was predicted and tested in vitro to achieve the highest ACE-I activity (IC50 = 0.16 μM), and the inhibition mechanism was also investigated. This study introduced a new method for predicting bio-activity with high precision using the 3D-QSAR model.

Release of an encrypted, highly potent ACE-inhibitory peptide by enzymatic hydrolysis of moth bean (Vigna aconitifolia) protein.

Dietary approaches for the regulation of blood pressure are the need of the hour. Hence, identifying the foods possessing such activity is gaining importance. With this aim, moth bean (Vigna aconitifolia), an underutilized pulse, was explored for the presence of antihypertensive activity in terms of angiotensin converting enzyme (ACE)-inhibition bioactivity. Defatted moth bean protein concentrate was hydrolyzed by using different proteases including Alcalase, papain, and trypsin, to identify the enzyme producing highly potent ACE inhibitory peptides. The hydrolysate showing the highest ACE inhibitory activity was further fractionated using an ultrafiltration membrane (10, 3 and 1 kDa) based on ACE inhibitory activity. The active fraction was further subjected to the ion-exchange chromatography followed by RP-HPLC and LC-MS/MS analysis for the enrichment and identification of ACE inhibitory peptides. Finally, based on the bioinformatic analysis, few peptides were synthesized and evaluated for ACE inhibitory activity, followed by docking study and molecular dynamic simulation of a peptide with the highest ACE inhibitory activity. Out of the three proteases, Alcalase-derived hydrolysate showed the highest (~59%) ACE inhibition activity. Molecular weight-based fractionation revealed that <1 kDa fraction possessed the highest ACE inhibitory activity. Activity guided separation of 1 kDa fraction using ion-exchange chromatography, RP-HPLC and LC-MS/MS showed the presence of about 45 peptides. Based on the bioinformatic analysis, 15 peptides were synthesized and evaluated for ACE inhibitory activity. Among these, a novel octapeptide FPPPKVIQ showed the highest ACE inhibitory activity (93.4%) with an IC50 of 0.24 μM. This peptide retained about 59% activity post gastrointestinal digestion simulation. A Dixon plot as well as docking studies revealed the uncompetitive inhibitory nature of this peptide with a Ki value of 0.81 μM. Molecular dynamic simulation studies till 100 ns ensured the stability of the ACE-peptide complex. Thus, present study identified a novel potent ACE inhibitory peptide from moth bean that can be incorporated in a functional dietary formulation for regulation of hypertension.

Interaction mechanism of egg derived peptides RVPSL and QIGLF with dipalmitoyl phosphatidylcholine membrane: microcalorimetric and molecular dynamics simulation studies.

Egg-derived peptides are becoming increasingly popular due to their biological activity and non-toxic effects. The egg-derived peptides Arg-Val-Pro-Ser-Leu (RVPSL) and Gln-Ile-Gly-Leu-Phe (QIGLF) display strong angiotensin-converting enzyme inhibitory activity and they can be taken up by intestinal epithelial cells. The interaction of the egg-derived peptides RVPSL and QIGLF with the membrane remains unclear. The position and structure of the peptides in the membrane were calculated. The maximum density values of RVPSL and QIGLF were 2.27 and 1.22 nm from the center of the 1,2-dipalmitoyl-sn-glycero-3-phosphatidylcholine (DPPC) membrane, respectively, indicating that peptides penetrated the membrane-water interface and were embedded in the membrane. The interaction of RVPSL and QIGLF with the DPPC membrane did not affect the average area per lipid or the lipid sequence parameters. The thermodynamic parameters ΔH, ΔG, and ΔS of the interaction between the peptide RVPSL with the DPPC membrane were 17.91 kJ mol-1 , -17.63 kJ mol-1 , 187.5 J mol-1 ·k-1 , respectively. The thermodynamic parameters ΔH, ΔG, and ΔS of the interaction between peptide QIGLF with DPPC membrane were 17.10 kJ mol-1 , -17.12 kJ mol-1 , 114.8 J mol-1 ·k-1 , respectively. The results indicated that the binding of peptides RVPSL and QIGLF to DPPC is an endothermic, spontaneous, and entropy-driven reaction. The results of the study are relevant to the problem of the low bioavailability of bioactive peptides (BP). © 2023 Society of Chemical Industry.

In vitro study on antidiabetic and antihypertensive activities of ethanolic extract of propolis of Indonesian stingless bee Tetragonula sapiens

The stingless bee, Tetragonula sapiens, is one of the species producing propolis in Indonesia. To date, there is still limited information about the standard characteristics and biological activity of propolis. This study investigated the standardized extract of Indonesian propolis in terms of its potency for controlling diabetes and high blood pressure in vitro, in accordance with its standard parameters. Three types of extracts; i.e., ethanolic extract of propolis (EEP), spray-dry propolis without wax (SDPNW), and spray-dry propolis with wax (SDPW); were prepared from T. sapiens stingless bees collected in South Sulawesi, Indonesia. The ethanolic extractable, loss on drying, ash, and water content parameters of EEP, SDPNW, and SDPW met the natural extract requirement. There is no significant value for their residual solvent levels (<0.54%), heavy metals (the absence of As, Cd, Pb, and Hg contamination), or the absence of microbial contamination. ACE inhibition activity on captopril, EEP, SDPNW, and SDPW had IC50 values of 5.97, 115.06, 112.93, and 116.70 ppm, respectively. While the IC50 values of acarbose, SDPNW, EEP, and SDPW for antidiabetic activity of α-glucosidase inhibition were 53.34, 56.48, 60.09, and 68.28 ppm, respectively. The TFC and TPC content of SDPNW were higher than EEP and SDPW, with values of 1.59% and 1.93%, respectively. It can be said that Indonesian propolis extract meets the standards for an extract and has the potential to reduce diabetic and hypertensive diseases. Further preclinical and clinical studies are needed to develop standardized propolis extract as an anti-diabetic and antihypertensive agent.

Oenococcus oeni allows the increase of antihypertensive and antioxidant activities in apple cider

This study aimed to investigate the impact of the malolactic fermentation (MLF) carried out by Oenococcus oeni on antihypertensive and antioxidant activities in cider. The MLF was induced using three strains of O. oeni. The modification in phenolic compounds (PCs) and nitrogen organic compounds, antioxidant, and antihypertensive activities were determined after MLF. Among the 17 PCs analyzed caffeic acid was the most abundant compound and phloretin, (−)-epicatechin, and myricetin were detected only in malolactic ciders, however, (−)-epigallocatechin was not detected after MLF. The evaluation of nitrogen organic compounds revealed a drop in total protein concentration (from 17.58 to 14.00 mg N/L) concomitantly with a significant release of peptide nitrogen (from 0.31 to a maximum value of 0.80 mg N/L) after MLF. In addition, an extracellular proteolytic activity was evidenced in all MLF supernatants. The FRAP activity increased reaching a maximum of 120.9 μmol FeSO4/mL and the ABTS radical-scavenging activity increased until 6.8 mmol ascorbic acid/L. Moreover, the angiotensin I-converting enzyme inhibitory activity reached a maximum value of 39.8%. The MLF conducted by O. oeni in ciders enables the increase of interesting biological activities and this finding could constitute a valuable tool to add value to final product.

Mechanism of angiotensin-converting enzyme inhibitors in the treatment of dilated cardiomyopathy based on a protein interaction network and molecular docking.

Dilated cardiomyopathy (DCM) is a severe manifestation or intermediate stage of cardiovascular disease progression with a significantly poor prognosis. Based on a protein interaction network and molecular docking, the present study determined the genes and mechanism of action of angiotensin-converting enzyme inhibitors (ACEIs) in the treatment of DCM, providing a direction for future studies on ACEI drugs for DCM. This is a retrospective study. DCM samples and healthy controls were downloaded from the GSE42955 dataset, and the targets of the potential active ingredients were obtained from PubChem. Hub genes in ACEIs were analyzed by constructing network models and a protein-protein interaction (PPI) network using the STRING database and Cytoscape software. Molecular docking was performed using Autodock vina software. Twelve DCM samples and five control samples were finally included. A total of 62 intersected genes were obtained by intersecting the differentially expressed genes with six ACEI target genes. PPI analysis identified 15 intersecting hub genes from these 62 genes. Enrichment analysis showed that the hub genes were associated with T helper type 17 (Th17) cell differentiation as well as the nuclear factor kappa-B (NF-kappa B), interleukin 17 (IL-17), mitogen-activated protein kinase (MAPK), tumor necrosis factor (TNF), phosphatidylinositol 3 kinase (PI3K)/protein kinase B (AKT) (PI3K-Akt), and Toll-like receptor signaling pathways. Molecular docking indicated that the compound Benazepril to produce favorable interactions with TNF proteins with a relatively higher score (-8.3). This study primarily revealed that the preventive and curative effects of ACEI treatment on DCM could be realized through multiple targets and pathways, and its mechanism of action is related to genes such as TNF, vascular endothelial growth factor A (VEGFA), interleukin 6 (IL6), C-C motif chemokine ligand 2 (CCL2), Cyclin D1 (CCND1), and AKT serine/threonine kinase 1 (AKT1), with immune- and inflammation-related signaling pathways involvement.

ACE inhibitory activity and salt-reduction properties of umami peptides from chicken soup

Angiotensin-I-converting enzyme (ACE) inhibitory activity and salt-reduction properties of umami peptides identified in chicken soup were investigated. The ACE inhibition rate of TPLVDR (91.22%) and AEINKILGN (81.26%) was significantly higher than other umami peptides, and their semi-inhibitory concentration was 0.017 mM and 0.034 mM, respectively. After in vitro digestion, the inhibitory activity of AEINKILGN and TPLVDR decreased, but the original sequences were still detected. The docking results showed that AEINKILGN and TPLVDR mainly interacted with Zn2+ and key sites (His353, Lys511and Glu411) in the active pockets of ACE through hydrogen bonds, which was crucial to the ACE inhibitory activity. Based on response surface methodology and sensory analysis, saltiness and palatability models were established to investigate the salt-reduction effect. The optimal level of AEINKILGN was about 1.16 mg/mL in 0.44% salt solution. And the TPLVDR was applicable to the low salt solution (0.1–0.2%) at a concentration from 0.23 to 0.29 mg/mL.

ACE-Inhibitory Peptides Identified from Quinoa Bran Glutelin-2 Hydrolysates: In Silico Screening and Characterization, Inhibition Mechanisms of ACE, Coordination with Zinc Ions, and Stability.

To obtain Angiotensin-I-Converting Enzyme (ACE) inhibition peptides with Zn-chelating capacity, quinoa bran glutelin-2 hydrolysates (QBGH) by Flavourzyme and Papain were subjected to Sephadex G-15 gel chromatography, reverse phase-high liquid performance chromatography and UPLC-ESI-MS/MS analysis. Four oligopeptides including GGGSGH, EAGAE, AGGGAGGG and AVPKPS were identified. Of these, only the hexapeptide AVPKPS had both ACE-inhibitory activity (IC50: 123.13μmol/L) and Zn-chelating ability (17.36mg/g). Molecular docking showed AVPKPS could bind with active residues Glu384 and Ala354 (both belong to the central S1 pocket of ACE including) through short hydrogen bond and hydrophobic interactions, respectively. Inhibition kinetics verified that AVPKPS was a competitive inhibitor of ACE. Moreover, AVPKPS can affect the zinc tetrahedral coordination in ACE through binding with residues His387 and His383. Fourier-transform infrared spectroscopy analysis demonstrated that the amino and carboxyl groups of AVPKPS were the main chelating sites for zinc ions. Under the gastrointestinal digestion, the ACE inhibition capacity of AVPKPS was relatively stable, and the zinc solubility of AVPKPS-zinc complexes was more stable than zinc sulfate (p < 0.05). These results suggest that quinoa peptides have potential applications as ingredients for antihypertension or zinc fortification.

Integrated metabolomics and peptidomics to delineate characteristic metabolites in milk fermented with novel Lactiplantibacillus plantarum L3.

A novel wild-type Lactiplantibacillus plantarum (L. plantarum) L3 with good fermentation characteristics and protein degradation capacity was isolated from raw milk samples. In this study, the metabolites in milk fermented with L. plantarum L3 were investigated by metabolomic and peptidomics analyses. The metabolomics results revealed that the metabolites in milk fermented with L. plantarum L3 were Thr-Pro, Val-Lys, l-creatine, pyridoxine, and muramic acid, which improved the taste and nutritional qualities of the milk. Moreover, the water-soluble peptides derived from L3 fermented milk exhibited high antioxidant properties and angiotensin I-converting enzyme inhibitory (ACEI) activities. Additionally, 152 peptides were found using liquid chromatography-mass spectrometry (LC-MS/MS). Furthermore, endogenous enzymes secreted by L. plantarum L3 cleaved β- and α-casein to release six ACEI peptides (ACEIPs), nineteen antioxidant peptides (AOPs), and five antimicrobial peptides (AMPS). Overall, these findings could be valuable in improving the quality of fermented milk.

Probiotic, technological and health-related characteristics of lactobacilli isolated from breast milk.

Isolation and characterisation of lactobacilli from human milk and determination of their probiotic, technological and in vitro health-promoting properties with a view to their potential use in food fermentation. Seven lactobacilli isolates were obtained from human milk and identified as Lacticaseibacillus paracasei (isolates BM1-BM6) and Lactobacillus gasseri (BM7). The isolates were examined in vitro for their technological, probiotic and health-promoting potential. Overall, all isolates showed important technological properties based on the ability to grow in milk whey, a high to moderate acidification capacity and the absence of undesirable enzymatic activities. L. gasseri (BM7) differed from the L. paracasei isolates by the absence of several glycosidases and the inability to ferment lactose. Isolates L. paracasei BM3 and BM5 produced exopolysaccharides (EPS) from lactose. All isolates showed probiotic potential as they were tolerant to simulated gastrointestinal conditions, had high cell surface hydrophobicity, had not acquired resistance to relevant antibiotics and had no virulence characteristics. All L. paracasei showed high antimicrobial activity against various pathogenic bacteria and fungi, while L. gasseri showed a narrower spectrum of antimicrobial activity. All isolates showed health-promoting potential in vitro, as evidenced by high cholesterol-lowering activity, high ACE inhibitory activity and marked antioxidant activity. All strains showed excellent probiotic and technological properties for use in lactic ferments. Lactobacilli are an important part of the microbial population in human milk. A more detailed characterisation of these bacteria can contribute to the understanding of their functional and health importance for infants and to the selection of new probiotic strains for the dairy industry.

Identification and Characterization of Novel ACE Inhibitory and Antioxidant Peptides from Sardina pilchardus Hydrolysate.

Sardina pilchardus is a valuable source of bioactive peptides with potential applications in functional foods. In this study, we investigated the angiotensin-converting enzyme (ACE) inhibitory activity of Sardina pilchardus protein hydrolysate (SPH) produced using dispase and alkaline protease. Our results showed that the low molecular mass fractions (<3 kDa) obtained through ultrafiltration exhibited more effective ACE inhibition, as indicated by screening with ACE inhibitory activity. We further identified the low molecular mass fractions (<3 kDa) using an LC-MS/MS rapid screening strategy. A total of 37 peptides with potential ACE inhibitory activity were identified based on high biological activity scores, non-toxicity, good solubility, and novelty. Molecular docking was used to screen for peptides with ACE inhibitory activity, resulting in the identification of 11 peptides with higher -CDOCKER ENERGY and -CDOCKER INTERACTION ENERGY scores than lisinopril. The sequences FIGR, FILR, FQRL, FRAL, KFL, and KLF were obtained by synthesizing and validating these 11 peptides in vitro, all of which had ACE inhibitory activity, as well as zinc-chelating capacity. All six peptides were found to bind to the three active pockets (S1, S2, and S1') of ACE during molecular docking, indicating that their inhibition patterns were competitive. Further analysis of the structural characteristics of these peptides indicated that all six peptides contain phenylalanine, which suggests that they may possess antioxidant activities. After experimental verification, it was found that all six of these peptides have antioxidant activities, and we also found that the SPH and ultrafiltration fractions of SPH had antioxidant activities. These findings suggest that Sardina pilchardus may be a potential source of natural antioxidants and ACE inhibitors for the development of functional foods, and using LC-MS/MS in combination with an online database and molecular docking represents a promising, effective, and accurate approach for the discovery of novel ACE inhibitory peptides.

Enzymatic Hydrolysis of Tenebrio molitor (Mealworm) Using Nuruk Extract Concentrate and an Evaluation of Its Nutritional, Functional, and Sensory Properties.

Enzymatic protein hydrolysis is a well-established method for improving the quality of dietary proteins, including edible insects. Finding effective enzymes from natural sources is becoming increasingly important. This study used nuruk extract concentrate (NEC), an enzyme-rich fermentation starter, to produce protein hydrolysate from defatted Tenebrio molitor (also called mealworm, MW). The nutritional, functional, and sensorial properties of the hydrolysate were then compared to those obtained using commercial proteases (alcalase and flavourzyme). The protease activities of the crude nuruk extract (CNE), NEC, alcalase, and flavourzyme were 6.78, 12.71, 11.07, and 12.45 units/mL, respectively. The degree of hydrolysis and yield of MW hydrolysis by NEC were 15.10 and 35.92% (w/w), respectively. MW hydrolysate was obtained using NEC and had a significantly higher free amino acid content (90.37 mg/g) than alcalase (53.01 mg/g) and flavourzyme (79.64 mg/g) hydrolysates. Furthermore, the NEC hydrolysis of MW increased the antioxidant and angiotensin-converting enzyme inhibitory activity, with IC50 values of 3.07 and 0.15 mg/mL, respectively. The enzymatic hydrolysis also improved sensory properties, including umaminess, sweetness, and saltiness. Overall, this study found that the NEC hydrolysis of MW outperformed commercial proteases regarding nutritional quality, sensory attributes, and biological activity. Therefore, nuruk could potentially replace commercial proteases, lowering the cost of enzymatic protein hydrolysis.

Soilless Cultivated Halophyte Plants: Volatile, Nutritional, Phytochemical, and Biological Differences.

The use of halophyte plants appears as a potential solution for degraded soil, food safety, freshwater scarcity, and coastal area utilization. These plants have been considered an alternative crop soilless agriculture for sustainable use of natural resources. There are few studies carried out with cultivated halophytes using a soilless cultivation system (SCS) that report their nutraceutical value, as well as their benefits on human health. The objective of this study was to evaluate and correlate the nutritional composition, volatile profile, phytochemical content, and biological activities of seven halophyte species cultivated using a SCS (Disphyma crassifolium L., Crithmum maritimum L., Inula crithmoides L., Mesembryanthemum crystallinum L., Mesembryanthemum nodiflorum L., Salicornia ramosissima J. Woods, and Sarcocornia fruticosa (Mill.) A. J. Scott.). Among these species, results showed that S. fruticosa had a higher content in protein (4.44 g/100 g FW), ash (5.70 g/100 g FW), salt (2.80 g/100 g FW), chloride (4.84 g/100 g FW), minerals (Na, K, Fe, Mg, Mn, Zn, Cu), total phenolics (0.33 mg GAE/g FW), and antioxidant activity (8.17 µmol TEAC/g FW). Regarding the phenolic classes, S. fruticosa and M. nodiflorum were predominant in the flavonoids, while M. crystallinum, C. maritimum, and S. ramosissima were in the phenolic acids. Moreover, S. fruticosa, S. ramosissima, M. nodiflorum, M. crystallinum, and I. crithmoides showed ACE-inhibitory activity, an important target control for hypertension. Concerning the volatile profile, C. maritimum, I. crithmoides, and D. crassifolium were abundant in terpenes and esters, while M. nodiflorum, S. fruticosa, and M. crystallinum were richer in alcohols and aldehydes, and S. ramosissima was richer in aldehydes. Considering the environmental and sustainable roles of cultivated halophytes using a SCS, these results indicate that these species could be considered an alternative to conventional table salt, due to their added nutritional and phytochemical composition, with potential contribution for the antioxidant and anti-hypertensive effects.

Health-promoting activities of Moringa oleifera Lam. seeds protein hydrolysates and its ultra-filtered peptide fractions

Moringa oleifera (MO) seeds treated as a new resource food rich in protein (37.48%), but, little know the bioactive benefits and peptide composition of protein hydrolysates. In this study, ultrafiltrated peptide fractions of MO seed proteins hydrolysates (MOSPH) were analyzed for their antioxidant, anti-hypertension and sleep-promoting activities. LC-MS/MS was used to identify the peptide composition of the molecular weight (MW) <3 kDa fraction. The results showed that MOSPH and the lower MW peptides fraction, especially the MW <3 kDa fraction, exhibited higher free radical scavenging and reducing capacity, in vitro angiotensin I-converting enzyme (ACE) inhibitory capacity and vasodilatory and anti-hypertensive effects on spontaneously hypertensive rats (SHR) after the development of hypertension, sleep-promoting and anticonvulsant effects in male ICR mice. Furthermore, four potential bioactive peptides (pepsite score > 0.3) from MW <3 kDa fraction of MOSPH were explored via in silico approach, of which a novel dual-function peptide IWHHTFYNELR (IR-11) exhibited higher ACE inhibitory activity (IC50 = 1.057 mg/mL) and DPPH•-clearing activity (IC50 = 0.404 mg/mL). Molecular docking indicates that IR-11 interacted with ACE and DPPH• through hydrogen bonding and hydrophobic forces. Our results suggested that MO seed proteins peptides can be a potent functional agents for developing new functional food.

Hawaijar – An ethnic vegan fermented soybean food of Manipur, India: A comprehensive review

Hawaijar, ethnic vegan fermented soybean food of Manipur, India is culturally and gastronomically important indigenously produced food. It is alkaline, sticky, mucilaginous and slightly pungent and bears similar properties with many fermented soybean foods of Southeast Asia like natto of Japan, douchi of China, thua nao of Thailand, choongkook jang of Korea. The functional microorganism is Bacillus and has numerous health benefits like fibrinolytic enzyme, antioxidant, antidiabetic, and ACE inhibitory activities. It is also very rich in nutrients but unscrupulous production method and sale lead to food safety issues. Huge potential pathogen population upto the level of 107-10 cfu/g Bacillus cereus and Proteus mirabilis were detected. Recent studies revealed presence of enterotoxic and urease gene in microorganisms originated from hawaijar. Improved and regulated food chain will result in hygienic and safe hawaijar. It has scope for functional food and nutraceutical global market and hold potential to provide employment to enhance the overall socioeconomic status of the region. Scientific production of fermented soybean over the traditional methods is summarized in this paper along with food safety and health benefits. Microbiological aspects on fermented soybean along with nutritive values are critically explained inside the paper.

Effect of Deployment and Harvest Date on Growth and High-Value Compounds of Farmed Alaria esculenta.

Alaria esculenta is a brown seaweed farmed in many European countries for its biomass rich in useful bio compounds. This study aimed to identify the optimal growing season to maximise biomass production and quality. The seeded longlines of the brown seaweed were deployed in the southwest of Ireland in October and November 2019 and samples of the biomass were harvested in different dates, between March and June 2020. Biomass gain and composition, phenolic and flavonoid content (TPC and TFC) and biological activities (antioxidant and anti-hypertensive activities) of seaweed extracts prepared with Alcalase were evaluated. The biomass production was significantly higher for the line deployed in October (>20 kg·m-1). In May and June, an increasing amount of epiphytes was observed on the surface of A. esculenta. The protein content of A. esculenta varied between 11.2 and 11.76% and fat content was relatively low (1.8-2.3%). Regarding the fatty acids profile, A. esculenta was rich in polyunsaturated fatty acids (PUFA), especially in eicosapentaenoic acid (EPA). The samples analysed were very rich in Na, K, Mg, Fe, Mn, Cr and Ni. The content of Cd, Pb Hg was relatively low and below the maximum levels allowed. The highest TPC and TFC were obtained in extracts prepared with A. esculenta collected in March and levels of these compounds decreased with time. In general, the highest radical scavenging activities (ABTS and DPPH), as well as chelating activities (Fe2+ and Cu2+) were observed in early spring. Extracts from A. esculenta collected in March and April presented higher ACE inhibitory activity. The extracts from seaweeds harvested in March exhibited higher biological activity. It was concluded that an earlier deployment allows for maximising growth and harvest of biomass earlier when its quality is at the highest levels. The study also confirms the high content of useful bio compounds that can be extracted from A. esculenta and used in the nutraceutical and pharmaceutical industry.

Exploring the potential of katsuobushi grounds as a source of bioactive peptides through fermentation with Aspergillus sydowii

Dried bonito broth, made from katsuobushi flakes, is a commonly used seasoning in Japanese cuisine. While extracting the broth, high-quality proteins are left behind in the katsuobushi grounds, which could be a valuable source of bioactive substances via a hydrolysis process. The xerophilic Aspergillus sydowii showed potential for protein hydrolysis and production of amino acids and soluble peptides through solid-state fermentation. Several peptidases involved in katsuobushi protein hydrolysis were purified from the crude enzyme preparations and identified as aminopeptidase, trypsin-like serine protease, and elastinolytic serine protease. The hydrolysates from the culture contained glutamic acid, leucine, and lysine and markedly increased soluble peptides with molecular weights ranging from 0.2 to 17 kDa. Electronic tongue analysis indicated that the hydrolysates had substantially stronger salty and umami tastes. Moreover, the hydrolysates exhibited radical-scavenging activities, ferric ion reducing antioxidant power, and angiotensin converting enzyme inhibitory activity, as well as promoting growth in lactic acid bacteria. In addition, de novo peptide sequencing was performed for peptides fractionated from the hydrolysates, and eight peptides were identified. The hydrolysates produced by A. sydowii fermentation could have potential applications in the food industry as natural flavor enhancers and functional ingredients with several bioactivities.

Production and Characterization of ACE Inhibitory and Anti-Diabetic Peptides from Buffalo and Camel Milk Fermented with Lactobacillus and Yeast: A Comparative Analysis with In Vitro, In Silico, and Molecular Interaction Study.

The investigation aimed at assessing a comparative study on the production and characterization of ACE inhibitory, anti-diabetic, and anti-inflammatory activities, along with the production of ACE inhibitory and anti-diabetic peptides through the fermentation of buffalo and camel milk by Limosilactobacillus fermentum (KGL4) and Saccharomyces cerevisiae (WBS2A). The angiotensin-converting enzyme (ACE) inhibitory and anti-diabetic properties were evaluated at particular time intervals (12, 24, 36, and 48 h) at 37 °C, and we discovered maximum activity at 37 °C after 48 h of incubation. The maximum ACE inhibitory, lipase inhibitory activities, alpha-glucosidase inhibitory, and alpha-amylase inhibitory activities were found in the fermented camel milk (77.96 ± 2.61, 73.85 ± 1.19, 85.37 ± 2.15, and 70.86 ± 1.02), as compared to the fermented buffalo milk (FBM) (75.25 ± 1.72, 61.79 ± 2.14, 80.09 ± 0.51, and 67.29 ± 1.75). Proteolytic activity was measured with different inoculation rates (1.5%, 2.0%, and 2.5%) and incubation times (12, 24, 36, and 48 h) to optimize the growth conditions. Maximum proteolysis was found at a 2.5% inoculation rate and at a 48 h incubation period in both fermented buffalo (9.14 ± 0.06) and camel milk (9.10 ± 0.17). SDS-PAGE and 2D gel electrophoresis were conducted for protein purification. The camel and buffalo milk that had not been fermented revealed protein bands ranging from 10 to 100 kDa and 10 to 75 kDa, respectively, whereas all the fermented samples showed bands ranging from 10 to 75 kDa. There were no visible protein bands in the permeates on SDS-PAGE. When fermented buffalo and camel milk were electrophoresed in 2D gel, 15 and 20 protein spots were detected, respectively. The protein spots in the 2D gel electrophoresis ranged in size from 20 to 75 kDa. To distinguish between different peptide fractions, water-soluble extract (WSE) fractions of ultrafiltration (3 and 10 kDa retentate and permeate) of fermented camel and buffalo milk were employed in RP-HPLC (reversed-phase high-performance liquid chromatography). The impact of fermented buffalo and camel milk on inflammation induced by LPS (lipopolysaccharide) was also investigated in the RAW 264.7 cell line. Novel peptide sequences with ACE inhibitory and anti-diabetic properties were also analyzed on the anti-hypertensive database (AHTDB) and bioactive peptide (BIOPEP) database. We found the sequences SCQAQPTTMTR, EMPFPK, TTMPLW, HPHPHLSFMAIPPK, FFNDKIAK, ALPMHIR, IPAVFK, LDQWLCEK, and AVPYPQR from the fermented buffalo milk and the sequences TDVMPQWW, EKTFLLYSCPHR, SSHPYLEQLY, IDSGLYLGSNYITAIR, and FDEFLSQSCAPGSDPR from the fermented camel milk.

Soypeptide supplementation attenuates weight loss and improves quality of life among brain cancer patients in a pilot study

AbstractInability to meet nutrition needs resulting from multiple diseases‐related, and individual factors contribute significantly to malnutrition and poor disease outcome among cancer patients. Strategies capable of delivering metabolically efficient nutrients with less digestive and metabolic stress without adding bulk to the diet of patients may be suitable for dealing with cancer‐related malnutrition and quality of life (QoL). We sought to test the hypothesis that peptide supplementation could attenuate weight loss and improve QoL among brain cancer patients compared to conventionally treated controls. In a non‐blind pilot study, brain cancer patients received 9 g soybean peptide supplementation twice per day (peptide group, n = 9) and compared with conventional treatment (control group, n = 10) over two chemotherapy cycles of 42 days. Compared to the controls, soypeptide supplementation attenuated weight loss with 5.4% increase in weight. Moreover, QoL, assessed using the European Organization for Research and Treatment of Cancer QLQ‐C30, revealed that physical (8.1 ± 15.6, p &lt; .047) and emotional (7.5 ± 3.5, p &lt; .033) functioning as well as summary scores (7.1 ± 2.4, p &lt; .048) improved, whereas others remain stable. Molecular docking simulation suggested that the peptides could induce body weight through the inhibition of angiotensin‐converting enzyme (ACE) upon digestion. Daily soypeptide supplementation could enhance weight gain and improve QoL among brain cancer patients possibly by inhibiting ACE activity. However, a follow‐up study with a larger sample size in a double‐blind randomized trial is required to validate these preliminary findings.

Screening of protein hydrolysates from malt rootlets and their relationship with technological properties

The malting industry produces a large quantity of by-products, and these have barely been explored in the research into their valorisation. Malt rootlets are rich in protein, and that content can be extracted to generate ingredients with a range of functionalities. Our study confirms that the extraction conditions for the type of enzyme and concentration, as well as the temperature, can affect the production yield and properties from malt rootlets protein hydrolysates. The pH effect was also studied, and it has only shown an effect on technological properties. Alcalase® can generate protein hydrolysates with higher protein content, reaching 33.2%, which in turn implies a higher emulsifying activity (up to 33.9% greater). On the other hand, Protamex® ensures a higher weight yield (up to 49.8%) and higher emulsifying stability, reaching 53.6%. In any case, protein hydrolysates do not improve foaming capacity on malt rootlets mainly due to relative low values of total protein content needed for these properties. All protein hydrolysates showed a high angiotensin-converting enzyme (ACE) inhibitory activity attributed to the similar amount of low molecular fraction (500–200 Da) present in all samples.