Decoding the Duality of Antinutrients: Assessing the Impact of Protein Extraction Methods on Plant-Based Protein Sources.

This review aims to provide an updated overview of the effects of protein extraction/recovery on antinutritional factors (ANFs) in plant protein ingredients, such as protein-rich fractions, protein concentrates, and isolates. ANFs mainly include lectins, trypsin inhibitors, phytic acid, phenolic compounds, oxalates, saponins, tannins, and cyanogenic glycosides. The current technologies used to recover proteins (e.g., wet extraction, dry fractionation) and novel technologies (e.g., membrane processing) are included in this review. The mechanisms involved during protein extraction/recovery that may enhance or decrease the ANF content in plant protein ingredients are discussed. However, studies on the effects of protein extraction/recovery on specific ANFs are still scarce, especially for novel technologies such as ultrasound- and microwave-assisted extraction and membrane processing. Although the negative effects of ANFs on protein digestibility and the overall absorption of plant proteins and other nutrients are a health concern, it is also important to highlight the potential positive effects of ANFs. This is particularly relevant given the rise of novel protein ingredients in the market and the potential presence or absence of these factors and their effects on consumers' health.

A comparative study of the digestion behavior and functionality of protein from chia (Salvia hispanica L.) ingredients and protein fractions.

Protein derived from chia (Salvia hispanica L.), characterized by a balanced amino acid composition, represents a potentially healthier and environmentally friendly alternative poised for innovation within the plant-based food sector. It was hypothesized that the growing location of chia seeds and processing techniques used might influence protein digestion patterns, which in turn could affect the biological functions of the digestion products. To examine this hypothesis, we assessed the gastrointestinal fate of degummed-defatted flour (DDF), protein concentrate (PC), and isolated albumin (Alb) and globulin (Glo) fractions. Furthermore, we compared the antioxidant and anti-inflammatory activities of the resulting digesta by means of in vitro and cellular assays. Post-gastrointestinal digestion, the PC exhibited elevated levels of soluble protein (7.6 and 6.3 % for Mexican and British PC, respectively) and peptides (24.8 and 27.9 %, respectively) of larger molecular sizes compared to DDF, Alb, and Glo. This can be attributed to differences in the extraction/fractionation processes. Leucine was found to be the most prevalent amino acids in all chia digesta. Such variations in the digestive outcomes of chia protein components significantly influenced the bioactivity of the intestinal digestates. During gastrointestinal transit, British Glo exhibited the best reactive oxygen species (ROS) inhibition activity in oxidative-stressed RAW264.7 macrophages, while Mexican digesta outperformed British samples in terms of ROS inhibition within the oxidative-stressed Caco-2 cells. Additionally, both Mexican and British Alb showed effectively anti-inflammatory potential, with keratinocyte chemoattractant (KC) inhibition rate of 82 and 91 %, respectively. Additionally, Mexican PC and Alb generally demonstrated an enhanced capacity to mitigate oxidative stress and inflammatory conditions in vitro. These findings highlight the substantial potential of chia seeds as functional food ingredients, resonating with the shifting preferences of health-conscious consumers.

Protein Hydrolysates and Bioactive Peptides as Mediators of Blood Glucose-A Systematic Review and Meta-Analysis of Acute and Long-Term Studies.

Type 2 diabetes mellitus (T2DM) is a major public health concern associated with high mortality and reduced life expectancy. Since diabetes is closely linked with lifestyle, not surprisingly, nutritional intervention and increased physical activity could play a vital role in attenuating the problems related to diabetes. Protein hydrolysates (PHs) and their bioactive peptides (BP) have been shown to exert a wide range of biological effects, including antioxidative, antihypertensive, and in particular, hypoglycaemic activities. To better understand the efficacy of such interventions, a systematic review and meta-analysis of randomised controlled trials (RCTs) were performed concerning the influence of protein hydrolysates on glycaemic biomarkers in subjects with and without hyperglycaemia. Five different databases were used to search for RCTs. In total, 37 RCTs were included in the systematic review and 29 RCTs in the meta-analysis. The meta-analysis revealed a significant reduction in postprandial blood glucose response (PPGR) in normoglycaemic (-0.22 mmol/L; 95% CI -0.43, -0.01; p ≤ 0.05) and in hyperglycaemic adults (-0.88 mmol/L; 95% CI -1.37, -0.39; p ≤ 0.001) compared with the respective control groups. A meta-regression analysis revealed a dose-dependent response for PPGR following PH consumption in normoglycaemic adults, specifically for doses ≤ 30 g. The postprandial blood insulin responses (PPIR) were significantly higher after the ingestion of PHs in both the group with and the group without hyperglycaemia, respectively (23.05 mIU/L; 95% CI 7.53, 38.57; p ≤ 0.01 and 12.57 mIU/L; 95% CI 2.72, 22.41; p ≤ 0.01), compared with controls. In terms of long-term responses, there was a small but significant reduction in both fasting blood glucose (FBG) and fasting glycated haemoglobin (HbA1c) in response to PH compared with the control group (p < 0.05). The PHs significantly improved the parameters of glycaemia in adults and, hence, it may contribute to the management and regulation of the future risk of developing T2DM.

Current state of insect proteins: extraction technologies, bioactive peptides and allergenicity of edible insect proteins.

This review aims to provide an updated overview of edible insect proteins and the bioactivity of insect-derived peptides. The essential amino acid content of edible insects is compared with well-known protein sources to demonstrate that edible insects have the potential to cover the protein quality requirements for different groups of the population. Then the current methodologies for insect protein extraction are summarized including a comparison of the protein extraction yield and the final protein content of the resulting products for each method. Furthermore, in order to improve our understanding of insect proteins, their functional properties (such as solubility, foaming capacity, emulsifying, gelation, water holding capacity and oil holding capacity) are discussed. Bioactive peptides can be released according to various enzymatic hydrolysis protocols. In this context, the bioactive properties of insect peptides (antihypertensive, antidiabetic, antioxidant and anti-inflammatory properties) have been discussed. However, the allergens present in insect proteins are still a major concern and an unsolved issue for insect-based product consumption; thus, an analysis of cross reactivity and the different methods available to reduce allergenicity are proposed. Diverse studies of insect protein hydrolysates/peptides have been ultimately promoting the utilization of insect proteins for future perspectives and the emerging processing technologies to enhance the wider utilization of insect proteins for different purposes.

In silico analysis of the interaction of de novo peptides derived from Salvia hispanica with anticancer targets.

Cancer is one of the leading causes of death worldwide. Conventional cancer therapies are not selective to cancer cells resulting in serious side effects on patients. Thus, the need for complementary treatments that improve the patient's response to cancer therapy is highly important. To predict and evaluate the physicochemical characteristics and potential anticancer activity of the peptides identified from S. hispanica protein fraction <1 kDa through the use of in silico tools. Peptides derived from Salvia hispanica's protein fraction <1 kDa were identified and analyzed for the prediction of their physicochemical properties. The characterized peptide sequences were then submitted to a multi-criteria decision analysis to identify the peptides that possess the characteristics to potentially exert anticancer activity. Through molecular docking analysis, the potential anticancer activity of the Potentially Anticancer Peptide (PAP)-1, PAP-2, PAP-3, PAP-4, and PAP-5 was estimated by their binding interactions with cancer and apoptosis-related molecules. All five evaluated PAPs exhibited strong binding interactions (< -100 kcal/mol). However, PAP-3 showed the lowest binding free energies with several of the targets. Thus, PAP-3 shows potential to be used as a nutraceutical or ingredient for functional foods that adjuvate in cancer treatment. Conclusions: Through the molecular docking studies, the binding of the PAPs to target molecules of interest for cancer treatment was successfully simulated, from which PAP-3 exhibited the lowest binding free energies. Further in vitro and in vivo studies are required to validate the predictions obtained by the in silico analysis.Communicated by Ramaswamy H. Sarma.

Identification of antidiabetic peptides derived from in silico hydrolysis of three ancient grains: Amaranth, Quinoa and Chia.

The antidiabetic potential of bioactive peptides derived from simulated gastrointestinal digestion (SGID) of proteins present in amaranth quinoa and chia was evaluated using their bioactivity profile and theoretical interaction with DPP-IV and α-glucosidases. In silico SGID generated 52 different fragments with in vitro antidiabetic activity where fragments PW, PF, PPG, PM, SW, IW, SF, PP, PPL, PG, PY, VW and PL scored highly in bioactivity probability, with molecular weights ranging from 172.2 to 325.44 Da; positive bulkiness index and hydrophobicity (except PP and PY) and no toxic properties. Fragments IW and PW presented the lowest free energy values for enzymes DPP-IV, maltase-glucoamylase, pancreatic α-amylase and sucrase-isomaltase (-8.2, -7.5, -7.7 and -7.5 kcal/mol; and -7.8, -7.4, -8.2, -7.4 kcal/mol respectively) We can conclude that proteins from amaranth, quinoa and chia may be a good source of antidiabetic BP and may exert antidiabetic activity through the release of BP after digestion.

Profiling modifications in physicochemical, chemical and antioxidant properties of wild blackberry (Rubus sp.) during fermentation with EC 1118 yeast.

Mexico is an extensively diverse country with a wide variety of wild species of blackberries (Rubus spp.), which are rich in bioactive compounds, however, these fruits are underutilized. Fermentation is a process that transforms the chemical compounds of fruits and increases nutraceutical properties. This study aimed to determine the physicochemical changes and the bioactive compounds profile that take place during the fermentation of wild blackberries using yeast EC 1118 and to evaluate its relationship with antioxidant activity (AOx). The results indicated that after 96 h of fermentation the content of carbohydrates (56%), total phenolic compounds (37%), and anthocyanins (22%), decreased, respectively. The physicochemical parameters showed statistic differences (p ≤ 0.05) at the endpoint of fermentation. The diversity of fatty acids was increased (55%), compared with unfermented blackberries. The modification of carbohydrates, anthocyanins, catechin, gallic and ellagic acid profiles were also monitored performing chromatographic techniques. The AOx, determined by ORAC and DPPH assays, showed the highest results for ORAC at 96 h increased a 140.2%, while DPPH values enhanced a 36.6% at 48 h of bioprocessing. Strong positive correlations were found between fermentation time and DPPH values (r = 0.8131), between ORAC and gallic acid content (r = 0.8688), and between anthocyanin content and pH (r = 0.9126). The fermentation of wild blackberries with EC 1118 yeast represents an alternative for development and formulation of potential ingredients for functional foods. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at (10.1007/s13197-020-04953-x).

In vitro gastrointestinal digestion impact on stability, bioaccessibility and antioxidant activity of polyphenols from wild and commercial blackberries (Rubus spp.).

Gastrointestinal digestion (GID) is a physiological process that transforms the stability, bioaccessibility and antioxidant activity (AOX) of polyphenols from blackberries (Rubus spp.). This study aimed to investigate the effect of the INFOGEST® GID protocol on the phenolic stability, bioaccessibility and AOX of Mexican wild (WB) and commercial (CB) blackberries. After GID, the total phenolic and anthocyanin contents in blackberries decreased by ≥68% and ≥74%, respectively. More than 40 phenolics were identified during GID; most of them degraded completely during digestion. GID had a negative effect on the AOX of both fruits (>50%), but WB showed the highest antioxidant activities, as assessed by the ORAC, DPPH, reducing power and ß-carotene bleaching methods. In Caco-2 cells, the cell-based antioxidant activity of digested blackberries (p < 0.05) decreased by 48% in WB and by 56% in CB. The capacity to inhibit intracellular ROS decreased by 50% in WB and by up to 86% in CB, after digestion. GID is a complex process that impacts on the bioactive properties of food nutrients, especially phenolics. In vitro and cellular AOX of WB polyphenols withstood the gastrointestinal environment better than CB phenolics. The in vitro assays results suggest that phenolics from underutilized WB have a higher bioaccessibility and antioxidant capacity than the polyphenols from the most frequently consumed CB. However, whether this corresponds to a better bioaccessibility in humans remains to be determined in future work.

Anticancer activity of protein fractions from chia (Salvia hispanica L.).

Salvia hispanica demonstrated to be a source of protein fractions with anticancer activity. The effect of the protein fractions <1, 1-3, and 3-5 kDa, obtained by ultrafiltration of the S. hispanica hydrolysate, was evaluated on the cellular viability of four cancer cell lines (MCF-7, Caco2, PC-3, and HepG2) and on human fibroblasts (hFB) at different concentrations (0.25, 0.5, 0.75, and 1 mg/ml). The protein fractions did not show cytotoxic effects on hFB. The protein fraction <1 kDa at 1 mg/ml showed the highest statistical effect on the cellular viability of all evaluated cancer lines; thus, its amino acid sequence was analyzed. From the multicriteria decision analysis of the peptide sequences obtained by mass spectrometry, the peptide KLKKNL with potential anticancer activity was selected. In conclusion, protein fractions could represent a therapeutic option for cancer treatment. However, further investigations are necessary to establish conclusive arguments. PRACTICAL APPLICATION: The work of this article is based on the background of the increasing potential of peptides for the treatment of chronic diseases. The results of this study present peptides of low molecular weight, obtained from chia seeds, as a potential adjuvant option for cancer treatment.

Impact of in vitro gastrointestinal digestion on peptide profile and bioactivity of cooked and non-cooked oat protein concentrates.

Oat (Avena sativa) is one of the most cultivated and consumed cereals worldwide. Recognized among cereals for its high protein content (12%-24%), it makes it an excellent source of bioactive peptides, which could be modified during processes such as heating and gastrointestinal digestion (GID). This work aims to evaluate the impact of heat treatment on the proteolysis of oat proteins and on the evolution of antioxidant peptide released during in vitro static GID, in terms of comparative analysis between cooked oat protein concentrate (COPC) and non-heated oat protein concentrate (OPC) samples. The protein extraction method and cooking procedure used showed no detrimental effects on protein quality. After GID, the proportion of free amino acids/dipeptides (<0.2 â€‹kDa) reached >40% for both samples (OPC and COPC), thus producing peptides with low molecular weight and enhanced bioactivity. Furthermore, during GID, the amino acid profile showed an increase in essential, positively-charged, hydrophobic and aromatic amino acids. At the end of GID, the reducing power of OPC and COPC increased >0.3 and 8-fold, respectively, in comparison to the non-digested samples; while ABTS•+ and DPPH• showed a >20-fold increase. Fe2+ chelating capacity of OPC and COPC was enhanced >4 times; similarly, Cu2+ chelation showed a >19-fold enhancement for OPC and >10 for COPC. ß-carotene bleaching activity was improved 0.8 times in OPC and >9 times in COPC; the oxygen radical antioxidant capacity assay increased 2 times in OPC and >4.7 times in COPC, respectively. This study suggests that OPC after cooking and GID positively influenced the nutritional and bioactive properties of oat peptides. Thus, COPC could be used as a functional food ingredient with health-promoting effects, as hydrothermal treatment is frequently used for this type of cereals.

Starch Digestion Enhances Bioaccessibility of Anti-Inflammatory Polyphenols from Borlotti Beans (Phaseolus vulgaris).

The consumption of beans has been associated with chronic disease prevention which may be attributed to the polyphenols present in the seed coat and endosperm. However, their bioaccessibility is likely to be limited by interactions with bean matrix components, including starch, protein and fibre. The aim of this project was to evaluate the effect of domestic processing and enzymatic digestion on the bioaccessibility of polyphenols from Borlotti beans (Phaseolus vulgaris) and to test their anti-inflammatory properties in a macrophage cell model. In vitro digestion of cooked beans released twenty times more polyphenols (40.4 ± 2.5 mg gallic acid equivalents (GAE)/g) than domestic processing (2.22 ± 0.1 mg GAE/g), with starch digestion contributing to the highest release (30.9 ± 0.75 mg GAE/g). Fluorescence microscopy visualization of isolated bean starch suggests that polyphenols are embedded within the granule structure. LC-MS analysis showed that cooked Borlotti bean contain flavonoids, flavones and hydroxycinnamic acids, and cooked bean extracts exerted moderate anti-inflammatory effects by decreasing mRNA levels of IL1ß and iNOS by 25% and 40%, respectively. In conclusion, the bioaccessibility of bean polyphenols is strongly enhanced by starch digestion. These polyphenols may contribute to the health benefits associated with bean consumption.

Malva species: Insights on its chemical composition towards pharmacological applications.

Malvaceae family is typical from the Mediterranean region, contains 240 genera and more than 4,200 species. They are most commonly used as ornamental plants, although they can also be conceived as a food resource and remedy for various diseases, such as digestive, respiratory, genitourinary, throat infections, and skeletal and skin disorders, as also injuries where they are profoundly applied for skin care and as antiseptic and demulcent. They also possess diuretic, lenitive, spasmolytic, and laxative effects, besides to be used as antidiarrheal. Thus, the present review provides in-depth data on Malva spp. potential applications and phytochemical composition for food and pharmaceutical industries. Habitat and cultivation conditions and the clinical reports related to its biological effects are also emphasized. Malva spp. possess a wide variety of chemical constituents (such as polysaccharides, coumarins, flavonoids, polyphenols, vitamins, terpenes, and tannins) found in different plant organs, especially in leaves and flowers, connected to their biological activity. In general, Malva spp. have rather moderate antimicrobial activity, high antiinflammatory and wound healing activities, strong antioxidant activity, and anticancer properties. Results from in vitro and in vivo experiments encourage more in-depth studies, namely clinical trials, towards to improve knowledge on the use of Malva spp. for the treatment of various health conditions in humans.

Encapsulation of Lactobacillus fermentum K73 by Refractance Window drying.

The purpose of this work was to model the survival of the microorganism and the kinetics of drying during the encapsulation of Lactobacillus fermentum K73 by Refractance Window drying. A whey culture medium with and without addition of maltodextrin were used as encapsulation matrices. The microorganism with the encapsulation matrices was dried at three water temperatures (333, 343 and 353 K) until reaching balanced moisture. Microorganism survival and thin layer drying kinetics were studied by using mathematical models. Results showed that modified Gompertz model and Midilli model described the survival of the microorganism and the drying kinetics, respectively. The most favorable process conditions found with the mathematical modelling were a drying time of 2460 s, at a temperature of 353 K. At these conditions, a product with 9.1 Log CFU/g and a final humidity of 10% [wet basis] using the culture medium as encapsulation matrix was obtained. The result shows that Refractance Window can be applied to encapsulate the microorganism probiotic with a proper survival of the microorganism.

Edible Insects Processing: Traditional and Innovative Technologies.

Insects are part of the human diet in many parts of the world. Their nutritional value is widely recognized. Currently, most edible insects are harvested from the wild, although semi-domestication and indoor farming have increased insect availability and the sustainability of production. In traditional cultures, insects are processed in a number of ways (steaming, roasting, smoking, frying, stewing, and curing, among others) to improve their sensory and nutritional qualities as well as their shelf-life. In order to increase consumer interest in the West, various technologies have been developed that are aimed primarily at using insects as ingredients in a non-recognizable form, such as powders or flour. These technologies include drying (sun-drying, freeze-drying, oven-drying, fluidized bed drying, and microwave-drying) and new processing methods (ultrasound-assisted extraction, cold atmospheric pressure plasma, and dry fractionation) designed mainly for protein, fat, and/or chitin extraction. Insect-based ingredients are sold for the production of cookies, chocolates, tortilla-style chips, and other snacks. This review focuses on edible insect production, processing technologies, and commercialization using strategies ranging from traditional to novel as a sustainable approach for improving food security worldwide.

Angiotensin-converting enzyme-inhibitory activity in protein hydrolysates from normal and anthracnose disease-damaged Phaseolus vulgaris seeds.

BACKGROUND: Bean seeds are an inexpensive source of protein. Anthracnose disease caused by the fungus Colletotrichum lindemuthianum results in serious losses in common bean (Phaseolus vulgaris L.) crops worldwide, affecting any above-ground plant part, and protein dysfunction, inducing the synthesis of proteins that allow plants to improve their stress tolerance. The aim of this study was to evaluate the use of beans damaged by anthracnose disease as a source of peptides with angiotensin-converting enzyme (ACE-I)-inhibitory activity. RESULTS: Protein concentrates from beans spoiled by anthracnose disease and from regular beans as controls were prepared by alkaline extraction and precipitation at isolelectric pH and hydrolysed using Alcalase 2.4 L. The hydrolysates from spoiled beans had ACE-I-inhibitory activity (IC(50) 0.0191 mg protein mL(-1)) and were very similar to those from control beans in terms of ACE-I inhibition, peptide electrophoretic profile and kinetics of hydrolysis. Thus preparation of hydrolysates using beans affected by anthracnose disease would allow for revalorisation of this otherwise wasted product. CONCLUSION: The present results suggest the use of spoiled bean seeds, e.g. anthracnose-damaged beans, as an alternative for the isolation of ACE-I-inhibitory peptides to be further introduced as active ingredients in functional foods.

Antioxidant and metal chelating activities of peptide fractions from phaseolin and bean protein hydrolysates.

Bean protein isolate and phaseolin were hydrolysed using pepsin and pancreatin, and the resulting hydrolysates were filtered through a 1kDa cut-off membrane and fractionated by size exclusion chromatography. Three fractions corresponding to MW 0.7-1.0kDa, 0.43-0.7kDa and <0.43kDa (A1, A2, and A3 for protein isolate fractions, and B1, B2, and B3 for phaseolin fractions) were assayed for antioxidant and metal chelating activity and they were also subjected to amino acid and SDS-PAGE analysis. Fractions A1 and B1 had the highest copper chelating activity (78% and 82%, respectively), while iron chelating activity was the highest in fractions A1 and B3 (36% and 16%, respectively). Fractions A2 and B3 had the highest antioxidant activity as determined by inhibition of reducing power and ß-carotene bleaching, while the highest ABTS radical scavenging activity was found in A3 and B3. Thus, fractions coming from the isolate and phaseolin had similar activities except for iron chelation, suggesting that phaseolin is the major contributor to the antioxidant and copper chelating activities of the hydrolysed protein isolate.