Impact of Alcalase Hydrolysis and Simulated Gastrointestinal Digestion on the Release of Bioactive Peptides from Erythrina edulis (Chachafruto) Proteins.

Amidst increasing awareness of diet-health relationships, plant-derived bioactive peptides are recognized for their dual nutritional and health benefits. This study investigates bioactive peptides released after Alcalase hydrolysis of protein from chachafruto (Erythrina edulis), a nutrient-rich South American leguminous plant, focusing on their behavior during simulated gastrointestinal digestion. Evaluating their ability to scavenge radicals, mitigate oxidative stress, and influence immune response biomarkers, this study underscores the importance of understanding peptide interactions in digestion. The greatest contribution to the antioxidant activity was exerted by the low molecular weight peptides with ORAC values for the <3 kDa fraction of HES, GD-HES, and GID-HES of 0.74 ± 0.03, 0.72 ± 0.004, and 0.56 ± 0.01 (µmol TE/mg protein, respectively). GD-HES and GID-HES exhibited immunomodulatory effects, promoting the release of NO up to 18.52 and 8.58 µM, respectively. The findings of this study highlighted the potential of chachafruto bioactive peptides in functional foods and nutraceuticals, supporting human health through dietary interventions.

Bioaccessibility and bioavailability of essential and potentially toxic trace elements in potato cultivars: A comprehensive nutritional evaluation.

Among the most consumed foods in the world is potato, which occupies the first place as a non-grain commodity, demonstrating the importance of its assessment concerning the population's food safety. In this study, the nutrients Ca, Mg, K, P, Cu, Mn, Fe, and Zn and the potentially toxic trace elements Cd, Cr, and Pb were evaluated considering their total contents, bioaccessible and bioavailable fractions in different potato cultivars, in an unpublished approach in the literature. The in vitro standard gastrointestinal digestion method (INFOGEST) and a model of the intestinal epithelial barrier using the Caco-2 cell line were applied for investigate the presence of metals in potato. For the macroelements, the bioaccessibility (% w/w) varied in the ranges: K (57-72 %), P (59-76 %), Mg (83-103 %), and Ca (30-123 %), whereas for the microelements were: Cu (27-74 %) and Mn (4.22-12.02, 60-119 %). The potentially of trace toxic elements, Cd and Pb, were found in 75 % of the samples, however, all the concentration values were below the maximum levels allowed of 0.10 µg/g. Chromium was determined only in potato peels and has no maximum established level. The bioaccessible and bioavailable fractions of Cd, Cr, and Pb were below the limits of quantification of the spectrometric methods (LOQ - µg/L: 0.063 Cd, 0.65 Cr, and 0.44 Pb). The potato samples were considered safe for consumption regarding the presence of potentially toxic trace elements, with a remarkable nutritional contribution.

Polyphenolic compounds of Phyllanthus amarus Schum & Thonn. (1827) and diabetes-related activity of an aqueous extract as affected by in vitro gastrointestinal digestion.

ETHNOPHARMACOLOGICAL RELEVANCE: Extracts of the aerial part of Phyllanthus amarus have been extensively used in several countries to cure diabetes. No data is available on the impact of gastrointestinal digestion of such crude extracts on their antidiabetic activity. AIM OF THE STUDY: The aim of this study was to identify active fractions and compounds of fresh aerial parts of P. amarus extracted by an infusion method that are responsible for antidiabetic effects occurring at the level of glucose homeostasis. MATERIALS AND METHODS: An aqueous extract was obtained by an infusion method and its polyphenolic composition was analysed by reverse phase UPLC-DAD-MS. The influence of in vitro gastrointestinal digestion was evaluated both on the chemical composition and on the antidiabetic effect of P. amarus infusion extract using glucose-6-phosphatase enzyme inhibition and stimulation of glucose uptake. RESULTS: Analysis of the chemical composition of the crude extract revealed the presence of polysaccharides and various families of polyphenols such as phenolic acids, tannins, flavonoids and lignans. After simulated digestion, the total content of polyphenols decreased by about 95%. Caffeoylglucaric acid derivates and lignans exhibited strong stimulation of glucose uptake similar to metformin with an increase of 35.62 ± 6.14% and 34.74 ± 5.33% respectively. Moreover, corilagin, geraniin, the enriched polysaccharides fraction and the bioaccessible fraction showed strong anti-hyperglycemic activity with about 39-62% of glucose-6-phosphatase inhibition. CONCLUSION: Caffeoylglucaric acid isomers, tannin acalyphidin M1 and lignan demethyleneniranthin were reported for the first time in the species. After in vitro gastroinstestinal digestion, the composition of the extract changed. The dialyzed fraction showed strong glucose-6-phosphatase inhibition.

Iridoids and polyphenols from chilean Gaultheria spp. berries decrease the glucose uptake in Caco-2 cells after simulated gastrointestinal digestion.

Berries are rich food sources of potentially health-beneficial (poly)phenols. However, they may undergo chemical modifications during gastrointestinal digestion. The effect of simulated gastrointestinal digestion on the content and composition of secondary metabolites from Gaultheria phillyreifolia and G. poeppigii berries was studied. The influence of the digested extracts on the in vitro metabolism and absorption of carbohydrates was evaluated. After simulated digestion, 31 compounds were detected by UHPLC-DAD-MS. The total content of anthocyanins decreased by 98-100%, flavonols by 44-56%, phenylpropanoids by 49-75% and iridoids by 33-45%, the latter showing the highest stability during digestion. Digested extracts inhibited α-glucosidase (IC50 2.8-24.9 µg/mL) and decreased the glucose uptake in Caco-2 cells by 17-28%. Moreover, a decrease in the mRNA expression of glucose transporters SGLT1 (38-92%), GLUT2 (45-96%), GLUT5 (28-89%) and the enzyme sucrase-isomaltase (82-97%) was observed. These results show the effect of simulated gastrointestinal digestion on the content and composition of Gaultheria berries.

Assessment of the bacterial diversity of agave sap concentrate, resistance to in vitro gastrointestinal conditions and short-chain fatty acids production.

Low bacterial diversity in the gut has been associated with the development of several diseases. Agave sap concentrate (ASC) is obtained from the thermal treatment of the fresh sap called "aguamiel", an artisanal Mexican food. In this study, we assessed the microbial diversity from three different ASC producing regions in Mexico using high-throughput sequencing of the 16S rRNA gene and evaluated their resistance to an in vitro gastrointestinal process as well as their ability to produce short-chain fatty acids (SCFA). Seven phyla and 120 genera were detected in ASC samples; Firmicutes had the highest relative read abundance at the phylum level, whereas Bacillus was the most abundant genus. Bacterial diversity at phylum and genus levels was highly dependent on the region where ASC was produced. The microbiota from a selected sample was resistant to low pH conditions, bile salts and intestinal enzymes. Moreover, bacteria were able to survive and grow in the colonic environment. SCFA production was comparable with that observed for a well-known probiotic, Lactobacillus plantarum 299v, that was used as control. These findings demonstrate that ASC contains a bacterial ecosystem with potential probiotic benefits.

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.

In vitro gastrointestinal digestion and cytotoxic effect of ovalbumin-conjugated linoleic acid nanocomplexes.

The aim of this work was to examine the behavior of conjugated linoleic acid (CLA) delivery systems based on ovalbumin (OVA) and their derived nanoparticles (OVAn1 and OVAn2), under static in vitro gastrointestinal digestion model. In addition, potential cytotoxic effect of these inclusion complexes on a human colon cancer cell line (HT-29) was evaluated. OVA was resistant to gastric and intestinal digestion, while OVA nanoparticles were very susceptible to digestive enzymes hydrolysis. Particle size distribution (PDS) and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) for OVA evidenced the presence of a protein fragment of similar size after simulated digestive process. Conversely, for nanoparticles, partial and total hydrolysis in gastric and intestinal phases, respectively, was evidenced. After in vitro gastrointestinal digestion, released CLA (RCLA) was assayed. In case of OVA, as CLA carrier, RCLA was 37%, while for OVA nanoparticles, lower RCLA values (~10-20%) were obtained. From cytotoxic assays, it was observed that CLA molecule was responsible for cell death, whereas OVA or their derived nanoparticles were not cytotoxic on HT-29 cells. On the other hand, flow cytometry analysis revealed that main death mechanism for CLA, and their inclusion complexes was apoptosis. OVA-CLA and OVAn1-CLA inclusion complexes displayed the highest potential cytotoxic activity and apoptotic index. Information derived from this work could be relevant for the design of CLA delivery systems as promising nanosupplements for production of new functional and excipient foods for both prevention and control of colon cancer.

Effect of drying methods on the gastrointestinal fate and bioactivity of phytochemicals from cocoa pod husk: In vitro and in silico approaches.

Cocoa pod husk (CPH) contains many nutraceutical phytochemicals whose gastrointestinal fate and bioactivity can be affected by drying methods. Microwave (MW), forced-air drying (AF), and AF plus extrusion (AF-E) dried CPH samples were chemically characterized, and their phenolic and theobromine (THB) contents were evaluated under oral-gastric-intestinal (in vitro) and colonic fermentation (ex vivo). Absorption, distribution, metabolism, excretion, and toxicity (ADEMT) properties of CPH's small molecules were evaluated in silico. The chemical composition of CPH [mostly carbohydrates/insoluble dietary fiber], polyphenol [total polyphenols > condensed tannin (CT) > monomeric flavonoids] differed minimally among samples, except for THB content (AF/AF-E > MW) and antioxidant capacity (MW > AF/AF-E). Time- trend gastrointestinal (X3 behavior) and colonic bioaccessibility were AF/AF-E > MW, but phenolic acids, procyanidins, and THB fluctuated in a sample-specific fashion. In silico modeling showed that bioactives of CPH easily crossed the intestinal epithelium illustrating their bioaccessibility and, permeability. These bioactives can act as receptor ligands in a structure-dependent manner, suggesting their use as a functional ingredient.

Effect of the nixtamalization process on the protein bioaccessibility of white and red sorghum flours during in vitro gastrointestinal digestion.

Protein bioaccessibility is a major concern in sorghum (Sorghum bicolor L. Moench) due to potential interactions with tannins affecting its nutritional value. Technological treatments such as boiling or alkaline cooking have been proposed to address this problem by reducing tannin-protein interactions. This research aimed to evaluate the impact of nixtamalization in the protein bioaccessibility from two sorghum varieties (red and white sorghum) during in vitro gastrointestinal digestion. Nixtamalization increased protein bioaccessibility in the non-digestible fraction (NDF) (5.26 and 26.31% for red and white sorghum, respectively). However, cooking showed a higher permeation speed of protein from red sorghum flours at the end of the intestinal incubation (9.42%). The SDS-PAGE profile of the digested fraction (DF) at 90 min of intestinal incubation indicated that, for red sorghum, cooking allows the formation of α and γ-kafirins while nixtamalization increase α-kafirin release. Principal Components Analysis (PCA) showed the association between nixtamalization and dissociation of δα kafirin complexes and increased protein content in the digestible fraction. In silico interactions indicated the highest biding energies for (+)-catechin and kafirin fractions (ß-kafirin: -7.0 kcal/mol; γ-kafirin: -5.8 kcal/mol, and δ-kafirin: -6.8 kcal/mol), suggesting a minor influence of depolymerized proanthocyanidin fractions with sorghum proteins as a result of the nixtamalization process. In conclusion, nixtamalization increased the bioaccessibility of sorghum proteins, depolymerizing condensed tannins, and breaking protein-tannin complexes. Such technological process improves the nutrimental value of sorghum, supporting its inclusion in the human diet.

Impact of cooking and nixtamalization on the bioaccessibility and antioxidant capacity of phenolic compounds from two sorghum varieties.

Sorghum (Sorghum bicolor L. Moench) has been sparsely used as human food due to certain anti-nutritional factors such as tannins that reduce its digestibility, although the grain is an important source of bioactive compounds such as phenolic compounds (PCs). This study aimed to assess the impact of cooking and alkaline cooking (nixtamalization) on the bioaccessibility and antioxidant capacity of PCs of two sorghum varieties (white/red). Nixtamalization was the most effective procedure for the reduction of tannins (74.3%). Gallic acid proved to be the most bioaccessible PC (6359 µg/g). The total phenolics and condensed tannins correlated with the antioxidant capacity (ABTS/DPPH; R2: 0.30-0.43, p < 0.05). These results confirm the potential of thermal procedures to significantly modify the bioaccessibility of sorghum compounds, enhancing their concentrations and reducing anti-nutritional factors (tannins) while improving their antioxidant capacity.

Impact of in vitro gastrointestinal digestion on the bioaccessibility and antioxidant capacity of bioactive compounds from Passion fruit (Passiflora edulis) leaves and juice extracts.

The fruits consumption is highly associated with a lessening in the risk of the chronic noncommunicable diseases. Despite their content of bioactive compounds, physiological conditions might affect their bioaccessibility and biological potential. Hence, the purpose of this research was to evaluate the bioaccessibility and in vitro antioxidant capacity of bioactive compounds (polyphenols, ascorbic acid, and mono/oligosaccharides) from passion fruit (Passiflora edulis) juice and leaves extracts (PJE and PLE, respectively) during an in vitro gastrointestinal digestion. PLE exhibited the highest overall bioaccessibility of phenolics and mono/oligosaccharides. Ascorbic acid and kaempferol were the most bioaccessible (up to 12- and 13-fold, respectively) compounds and showed the highest net permeabilities (0.32-34.65 × 10-5 ). Chlorogenic acid, quercetin, and xylose showed the highest correlation with the antioxidant capacity (ABTS/DPPH methods). These results indicated the ability of digestion to improve bioaccessibility and antioxidant capacity of bioactives compounds from P. edulis extracts. PRACTICAL APPLICATIONS: Several international organizations, World Health Organization (WHO) among them, are actively encouraging an increase in the fruit and vegetable intake worldwide in response to the global rise in noncommunicable diseases, among other actions. Fruits and processed foods are increasingly used in the European food industry. Additionally, exotic and tropical fruits cannot be produced in the European Union, which opens up a variety of opportunities for our Colombian tropical fruits such as mango or exotic fruits such as passion fruit. Evaluating parameters such as bioaccessibility and intestinal permeability of these bioactives derived from both P. edulis juice and leaves extracts contributes with new evidence regarding their stability along the gastrointestinal tract and their consumption as a source of health-promoting compounds. The results of this research are an important achievement on the way to make the transition from basic to applied research. Informing the health benefits of this tropical fruit can stimulate its demand.

Antithrombotic Activity of Brewers' Spent Grain Peptides and their Effects on Blood Coagulation Pathways.

Antithrombotic activity of brewers' spent grain peptides before and after simulated gastrointestinal digestion and their effects on blood coagulation pathways were evaluated. Two hydrolysates were produced using sequential enzymatic systems: alkaline protease + Flavourzyme (AF) and neutral protease + Flavourzyme (PF). Simulation of gastrointestinal digestion of AF and PF hydrolysates was made using porcine pepsin and pancreatin enzymes, obtaining the corresponding digested samples: AFD and PFD, respectively. Peptides were fractionated by ultrafiltration using a 1 kDa cut-off membrane. Hydrolysates had peptides with medium and low molecular weights (2100 and 500 Da, respectively), and Glu, Asp, Leu, Ala, and Phe were the most abundant amino acids. Gastrointestinal digested hydrolysates presented high proportion of small peptides (~500 Da), and higher amount of Val, Tyr, and Phe than hydrolysates. Mass spectrum (HDMS Q-TOF) of AFD-ultrafiltered fraction <1 kDa exhibited peptides from 500 to 1000 Da, which are not present in AF. PFD showed the generation of new peptides from 430 to 1070 Da. All samples showed thrombin inhibitory activity. However, no effect was observed on prothrombin time. Peptides <1 kDa from hydrolysates and digested samples delayed thrombin and thromboplastin time respect to the control (~63%). Also the samples showed thrombin inhibitory activity at common pathway level. Thus, brewers' spent grain peptides exerted their antithrombotic activity by inhibiting the intrinsic and common pathways of blood coagulation. This is the first report to demonstrate that brewers' spent grain peptides are able to delay clotting time after simulated gastrointestinal digestion.

Bioaccessibility during In Vitro Digestion and Antiproliferative Effect of Bioactive Compounds from Andean Berry ( Vaccinium meridionale Swartz) Juice.

Berry consumption is associated with colorectal-cancer chemoprevention, but digestive conditions can affect this property. The bioaccessibility and apparent permeability coefficients of bioactive compounds from Andean Berry Juice (ABJ) after in vitro gastrointestinal digestion and colonic fermentation were analyzed. The antiproliferative effect of the fermented nondigestible fraction was evaluated against SW480 colon-adenocarcinoma cells. Gallic acid displayed the highest bioaccessibility in the mouth, stomach, small intestine, and colon. However, chlorogenic acid exhibited the highest apparent permeability coefficients (up to 1.98 × 10-4 cm/s). The colonic-fermentation fraction showed an increase of ≥50% antiproliferative activity against SW480 cells (19.32%, v/v), equivalent to those of gallic acid (13.04 µg/g), chlorogenic acid (7.07 µg/g), caffeic acid (0.40 µg/g), ellagic acid (7.32 µg/g), rutin (6.50 µg/g), raffinose (0.14 mg/g), stachyose (0.70 mg/g), and xylose (9.41 mg/g). Bioactive compounds from ABJ are bioaccessible through the gastrointestinal tract and colon fermentation, resulting in antiproliferative activity.

Bioaccessibility and antioxidant activity of free phenolic compounds and oligosaccharides from corn (Zea mays L.) and common bean (Phaseolus vulgaris L.) chips during in vitro gastrointestinal digestion and simulated colonic fermentation.

Corn (Zea mays L.) and common beans (Phaseolus vulgaris L.) are alternative suitable ingredients for snacks, because of their content of bioactive compounds such as phenolic compounds (PC) and oligosaccharides (OS). However, there is no information about the transformation of these compounds associated with food matrix during gastrointestinal digestion. Therefore, the objective of this work was to simulate the whole digestion process (mouth to colon) to estimate bioaccessibility and small intestine permeability of free PC and OS, and the antioxidant capacity of free PC. Digested nixtamalized corn-cooked common bean chips exhibited significant different quantities of free PC and OS, and higher antioxidant activity compared to methanolic extract. The free PC showed high values of apparent permeability coefficients (0.023-0.729×10-3), related with their absorption in the small intestine. Both free PC and OS were retained in the non-digestible fraction of chips (10.24-64.4%) and were able to reach the colon. Our results suggest the digestion potential to increase chip bioactive compounds and antioxidant activity. Additional studies are required to evaluate their in vivo effects.

Effects of in vitro digestion and in vitro colonic fermentation on stability and functional properties of yerba mate (Ilex paraguariensis A. St. Hil.) beverages.

Yerba mate (Ilex paraguariensis) is a plant that grows naturally in South America. From its leaves and thin stems different kinds of beverages are prepared (chimarrão, tererê and tea mate), all of them rich in bioactive substances. The aim of this study was to evaluate the influence of in vitro gastrointestinal digestion and colonic fermentation on the stability of the polyphenols and on the antioxidant, antimicrobial and antitumoral activities of the yerba mate beverages. The phenolic chromatographic profile revealed that both the in vitro digestion and the colonic fermentation caused a pronounced decrease in 3,5-O-dicaffeoylquinic acid and 5-O-caffeoylquinic acid in the preparations. However, 3-O-caffeoylquinic acid, 4-O-caffeoylquinic acid and salvianolic acid I were only barely affected in all preparations. Despite the decrease in the phytochemicals content, yerba mate beverages maintain their functional properties such as antioxidant, antibacterial and antitumoral activities.

Spent coffee grounds, an innovative source of colonic fermentable compounds, inhibit inflammatory mediators in vitro.

Spent coffee grounds (SCG), rich in dietary fiber can be fermented by colon microbiota producing short-chain fatty acids (SCFAs) with the ability to prevent inflammation. We investigated SCG anti-inflammatory effects by evaluating its composition, phenolic compounds, and fermentability by the human gut flora, SCFAs production, nitric oxide and cytokine expression of the human gut fermented-unabsorbed-SCG (hgf-NDSCG) fraction in LPS-stimulated RAW 264.7 macrophages. SCG had higher total fiber content compared with coffee beans. Roasting level/intensity reduced total phenolic contents of SCG that influenced its colonic fermentation. Medium roasted hgf-NDSCG produced elevated SCFAs (61:22:17, acetate, propionate and butyrate) after prolonged (24h) fermentation, suppressed NO production (55%) in macrophages primarily by modulating IL-10, CCL-17, CXCL9, IL-1ß, and IL-5 cytokines. SCG exerts anti-inflammatory activity, mediated by SCFAs production from its dietary fiber, by reducing the release of inflammatory mediators, providing the basis for SCG use in the control/regulation of inflammatory disorders. The results support the use of SGC in the food industry as dietary fiber source with health benefits.