Search Results (80)

Moringa oleifera seeds are rich in protein, yet their potential as plant-based protein in food remains underutilized. This study evaluated the extraction efficiency, composition, and techno-functional properties of moringa seed protein isolate (MSPI) using enzyme-assisted (EAE), ultrasonic-assisted (UAE), and microwave-assisted extraction (MAE) methods, compared to conventional alkaline extraction (CE). EAE was performed with viscozyme (2%, pH 8, 50 °C, 2 h) and papain (1%, pH 7, 50 °C, 1 h), UAE at 40% amplitude (20 kHz, 20 min), and MAE at 800 W (50 °C, 90 s). All methods significantly improved extraction yield (14.60–30.08%), protein content (80.47–86.61%), solubility (40.78–60.09% at pH 10), and techno-functional properties over CE. However, MAE slightly reduced solubility. Phytates (0.83–0.49 g/100 g) and trypsin inhibitor activity significantly decreased (4.48–1.92 U/mg). In vitro protein digestibility improved (p < 0.05) across all samples (88.11–93.81%), with hydrolysis patterns supporting the enhanced digestibility. Structural modifications were indicated by altered surface hydrophobicity and thermal properties. SDS-PAGE showed consistent major protein bands at 17, 25, and 48–63 kDa, with EAE showing reduced intensity at ~63 kDa. While UAE and MAE achieved high protein yield and purity, EAE offered the best balance of functionality and digestibility, making it the most promising method for producing high-quality MSPI. These findings are relevant for guiding the selection of extraction methods for MSPI recovery for food applications. Full article

A soybean meal is a key protein source in human foods and animal feed, yet its digestibility is constrained by endogenous trypsin inhibitors (TIs). Thermal processing is the mainstream tool for TI inactivation, but high-intensity heat treatments increase energy consumption and can potentially denature proteins, diminishing nutritional quality. Reducing the thermal input while maintaining nutritional quality is, therefore, a critical challenge. One promising strategy is the use of soybean cultivars bred for low-TI expression, which may allow for milder processing. However, the performance of these low-TI cultivars under reduced heat conditions remains unstudied. This study treated soybean samples under four different temperatures (60, 80, 100, and 121 °C) for 10 min and investigated the impact of heat treatment on TI concentration, in vitro protein digestibility, and nutritional properties of meals from a conventional high-TI variety (Glenn) and a novel low-TI variety (VT Barrack). Results showed that heat treatment at 100 °C significantly improved protein digestibility and lower TI concentrations in both varieties. A negative correlation was observed between protein digestibility and TI concentration in both soybean varieties. At 100 °C, the low-TI variety achieved 81.4% protein digestibility with only 0.6 mg/g TIs, whereas the high-TI variety required 121 °C to achieve comparable protein digestibility and a TI reduction. These findings highlight that low-TI soybeans can lower the necessary thermal treatment to 100 °C to minimize TIs while simultaneously preserving protein quality and cutting energy demand, offering a practical, cost-effective approach to producing higher-quality soybean meals. Full article

Balantioides coli is the only ciliate currently described as an intestinal parasite of humans, although it can also infect other animals, particularly pigs. Its in vitro cultivation remains challenging, and no axenic culture system is currently available. Cultures are initiated by adding small amounts of feces containing cysts or trophozoites to the culture medium. Implantation success is lower when starting from cysts, and the mechanisms and early events of excystation remain poorly understood. In this study, we describe the sequence of events involved in excystation and identify factors potentially important for culture establishment. Cysts were obtained from orangutan feces and genetically confirmed as B. coli. Only viable cysts, determined by trypan blue or methylene blue exclusion, were used. After artificial digestion with pepsin and trypsin, cysts were incubated at 28 °C for up to 72 h in DMEM supplemented with L-glutamine, yeast extract, fetal bovine serum, and starch granules. Excystation began with a fissure in the cyst wall, allowing for bacterial entry. This appeared to stimulate the trophozoites, the increased motility of which progressively weakened and ruptured the wall, allowing for their emergence. Wall rupture and bacterial entry were critical for activation., whereas starch type had no apparent influence. Excystation occurred within the first hours; otherwise, cysts degenerated. Full article

Angiotensin-converting enzyme 2 (ACE2) is a cell-surface receptor that helps the body regulate blood pressure and endocrine secretions. Transmembrane serine protease 2 (TMPRSS2) is a cell surface protein expressed mainly by endothelial cells of the respiratory and digestive tract, which participates in the cleavage of protein peptide bonds with serine as the active site. These two proteins have been studied to be highly associated with infection with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Soybean trypsin inhibitor (SBTI) has special bioactivities such as anticarcinogenic and anti-inflammatory functions, which can be widely used in functional foods or drugs. Our study involved in vitro and in vivo experiments to elucidate the effect of SBTI on SARS-CoV-2 host invasion. First, it was confirmed that being under 250 μg/mL of SBTI was not toxic to HepG2, HEK293T, and Calu-3 cells. The animal study administered SBTI to mice once daily for 14 days. In the lungs, liver, and kidneys, the histopathologic findings of the SBTI group were not different from those of the control group, but the expression of ACE2, TMPRSS2, and CD147 was reduced. Thus, our findings suggest that the inhibition of ACE2, TMPRSS,2 and CD147 proteins by SBTI shows promise in potentially inhibiting SARS-CoV-2 infection. Full article

The objective of this work was to evaluate the effects of three commercial diets for ornamental fish (BIOMAA Spiruflake^® (T1), Tetra TetraMin Tropical Flakes^® (T2), LOMAS tropical fish flakes^® (T3)) compared to a balanced feed for trout (Silver Cup TM (T4)) on the growth, survival, digestive enzyme activity, and intestinal and liver morphology of guppy (Poecilia reticulata) fry after 31 days of feeding. The highest growth was in T2 (0.101 ± 0.004 g) and T4 (0.084 ± 0.008 g) (p < 0.05). Fish fed with T1, T2, and T4 presented the greatest total length. T4 (98 ± 2.828%) and T1 (96 ± 5.656%) had the highest survival. Fish fed with T2 and T4 showed the best growth index values (AWG, SGR, FCR). T4 showed greater alkaline protease, trypsin, and chymotrypsin activity and better in vitro digestibility. The fish fed the T4 diet had the lowest intestinal fold height and the smallest hepatocyte diameter. A cheaper aquaculture diet (T4) significantly improved guppy juvenile nutrition compared to ornamental feeds. Further research should focus on developing species-specific ornamental diets for enhanced fish welfare. Full article

Ingestion is one of the main exposure routes of humans and animals to microplastics (MPs). During digestion, MPs can interact with both gastrointestinal enzymes and food proteins. This study investigated the adsorption of trypsin onto polypropylene (PP) and polyethylene terephthalate (PET) MPs, the influence of MPs on trypsin structure and activity, and the in vitro trypsin digestibility of bovine meat extract (BME) sarcoplasmic proteins and BME α-Gal-carrying allergens (α-GalA) in the presence of PP and PET MPs. Trypsin, BME and α-GalA proteins interact with MPs, resulting in the formation of a soft (SC) and hard (HC) corona. This interaction is dynamic, leading to the adsorption and desorption of protein through time. Trypsin adsorption onto MPs results in slight structural changes in the SC and bulk solution, while a trypsin fraction residing in the HC loses most of its specific activity. The presence of MPs slightly slows down the digestibility of proteins with a mass of 38 kDa, while it does not affect the digestion of α-GalA. According to our results, it is unlikely that realistic concentrations of MPs in the intestine would have significant effects on meat extract proteins’ and allergens’ digestibility by trypsin. We confirmed that during trypsin digestion, the corona on PP and PET MP is composed of BME sarcoplasmic proteins and allergenic α-Gal-carrying proteins. Full article

Legumes have been identified as a key element of food innovation and excellent candidates for ensuring sustainability in food systems. However, certain legumes, such as faba beans and legume by-products, such as pea pods, are currently mainly being used in animal feed rather than exploited and valued in human nutrition. In this study, the nutritional properties, anti-nutritional factors, and in vitro protein digestibility of pea pod flour and raw and thermally treated (80, 120, 150, and 180 °C during 30 min) faba bean flours were investigated. For pea pod flours, the results showed a very interesting protein content (12.13%) and insoluble fibers (37.45%), as well as appreciable amounts of minerals, mainly calcium, potassium, magnesium, manganese, and iron. For faba bean flours, thermal treatment did not significantly affect the crude protein, ash, starch, and fat contents of the processed beans. Meanwhile, compared with raw faba bean flours, thermal treatment significantly decreased insoluble dietary fibers, anti-nutritional factors such as phytic acid, tannins, trypsin inhibitors, and alpha-galactosides and progressively improved the in vitro protein digestibility by 7,7%. In conclusion, faba bean and pea pod flours show significant potential as novel ingredients in the food industry. Their combination will enable the development of protein, fiber, and mineral-rich food products. Full article

This study investigates the effects of fermentation on sainfoin seed flour using Saccharomyces boulardii for total dietary fiber (TDF) content, anti-nutritional profiles (including phytates, tannins, saponins, and trypsin inhibitors), and bioactive compounds. It also focused on assessing the in vitro availability of phenolic compounds, antioxidant potential, and anti-nutrient compounds after gastrointestinal digestion. Four treatment groups were designed: a non-fermented control group, and flour samples fermented with S. boulardii CNCM I-745 for 24, 48, and 72 h. All fermentations were carried out at 30 °C. The effects of fermentation and the analysis results were statistically evaluated at the significance level of p < 0.05, and significant differences were detected. Fermentation significantly increased soluble dietary fiber (from 3.32% to 4.43%) and reduced anti-nutritional factors, including phytates (by 18%), tannin (by 19%), and trypsin inhibitor activity (TIA) (by 79%). However, saponin content increased by 21% after 72 h of fermentation. Tannin levels of non-fermented and fermented sainfoin flour decreased dramatically after in vitro digestion. Moreover, it was concluded that the bioaccessibility of phytic acid significantly increased through fermentation, while that of tannins declined. Antimicrobial activity against Escherichia coli ATCC 25922 improved after fermentation, while the antioxidant capacity was enhanced post-digestion. In addition, the highest phenolic content (612 mg GAE/100 g) and antioxidant capacity (1745 mg TE/100 g by CUPRAC assay and 1127 mg TE/100 g by DPPH assay) were determined in fermented sainfoin seed flour at 72 h after gastrointestinal digestion. Full article

Squid pen (SP) was found to contain 64.41% protein and 26.03% chitin. The amino acid composition revealed that Met was the most abundant amino acid in SP, with a concentration of 13.67 g/100 g. To enhance the stability and bioavailability of SP hydrolysates, microcapsules were developed using ultrasonic emulsification techniques with SP trypsin hydrolysates (SPTH) and SP β-chitosan (SPC). The optimal preparation conditions involved using a 2% concentration of SPC, a 4 mg/mL concentration of SPTH, a core-to-wall ratio (v/v) of 1:3 for SPTH/SPC, and subjecting them to ultrasonic treatment for 20 min. These microcapsules had a loading capacity of 58.95% for SPTH under these conditions. The successful encapsulation of SPTH in the SPC complex to form SPC-SPTH microcapsules was confirmed by FTIR, XRD, DSC, and SEM, exhibiting good thermal stability, small particle size, and high encapsulation efficiency. In vitro digestion studies demonstrated a release of 15.61% in simulated gastric fluid and 69.32% in intestinal fluid, achieving targeted release in the intestines. The digested products exhibited superior antioxidant activity compared to free SPTH digests, suggesting that microencapsulation effectively preserves SPTH bioactivity. This study enhances the bioavailability of SPTH and offers a promising delivery system for natural compounds with low bioavailability and stability. Full article

Food processing can alter protein biochemical properties, impacting immunoreactivity and allergenicity. A key feature of food allergens is their resistance to enzymatic digestion, particularly by pepsin and trypsin. This study compares the digestomes of raw and heat- and/or pressure-treated peanuts, hazelnuts, pistachios and cashews using the INFOGEST harmonized digestion protocol and analyzing their IgE-binding capacity through in vitro methods. Protein patterns from controls and digestomes were resolved by SDS-PAGE and tested with sera from allergic patients, confirmed by competitive ELISA for hazelnuts and peanuts. The results indicate that processing methods differently affect the gastrointestinal (GI) digestion of these allergens. Simulated GI digestion caused a significant destruction of protein structures, reducing but not eliminating IgE reactivity for all four nuts. Boiling for 60 min did not change the SDS-PAGE profiles, but it did stimulate enzymatic activity, decreasing IgE binding capacity. In contrast, applying heat and pressure led to a nearly complete inhibition of allergenic potential during simulated digestion. These findings suggest that employing intense food processing techniques and investigating the gastrointestinal effects of highly allergenic nuts could be crucial steps toward developing new hypoallergenic formulations. Full article

The fall armyworm, Spodoptera frugiperda (Lepidoptera: Noctuidae), is a polyphagous insect of various agricultural crops. The methods used for its control have led to the selection of resistant insect populations, which justifies the search for new alternatives for the management of this insect. Accordingly, in the present study, trypsin inhibitors present in the leaf extract of Ricinus communis (Euphorbiaceae) were investigated for their activity against S. frugiperda. Chemometric optimization methods were developed for the extraction, purification, identification, and structural characterization of the inhibitors. In addition, the effect of R. communis extract on S. frugiperda development was evaluated. Inhibitor extraction was chemometrically optimized, yielding an extract with an antitryptic activity of 94,837.14 mUIT g⁻¹. The R. communis extract was purified and found to contain two compounds (adenosine and ricinine) exhibiting trypsin inhibitor activity. However, after purification, only ricinine inhibited S. frugiperda trypsin in vitro (103.21 mUIT mg⁻¹). The extract was added to the diet of S. frugiperda larvae, resulting in reduced digestion, increased protein in the feces (control = 12,571 μg protein/mg feces; 1818.2 g mL⁻¹ = 16,867 μg protein/mg feces), and insect mortality. At the highest concentration, the treatment led to an LT50 of 15.9 days and a cumulative survival rate of 18.5%. Based on the results of this study, it is possible that ricinine binds to the catalytic site of trypsin, causing the mortality of S. frugiperda larvae. Full article

The purpose of this study was to investigate the effect of micronization on the characteristics of black cumin pressing waste material. The basic composition, amino acid, and fatty acid content of the raw material—specifically, black cumin pressing waste material—were determined. The samples were micronized in a planetary ball mill for periods ranging from 0 to 20 min. The particle sizes of micronized samples of black cumin pressing waste material were then examined using a laser analyzer, the Mastersizer 3000. The structures of the produced micronized powders was examined by X-ray diffraction. Additionally, the FTIR (Fourier-transform infrared) spectra of the micronized samples were recorded. The measurement of phenolic and antiradical properties was conducted both before and after in vitro digestion, and the evaluation of protein digestibility and trypsin inhibition was also conducted. The test results, including material properties, suggest that micronization for 10 min dramatically reduced particle diameters (d50) from 374.7 to 88.7 µm, whereas after 20 min, d50 decreased to only 64.5 µm. The results obtained using FTIR spectroscopy revealed alterations, especially in terms of intensity and, to a lesser extent, the shapes of the bands, indicating a significant impact on the molecular properties of the tested samples. X-ray diffraction profiles revealed that the internal structures of all powders are amorphous, and micronization methods have no effect on the internal structures of powders derived from black cumin pressing waste. Biochemical analyses revealed the viability of utilizing micronized powders from black cumin pressing waste materials as beneficial food additives, since micronization increased total phenolic extraction and antiradical activity. Full article

Lima beans (Phaseolus lunatus) and adzuki beans (Vigna angularis) are some of the most nutritious underutilized pulses that are significant in being used as basic ingredients for the preparation of various food products. The present study aimed to determine the impact of soaking and germination on nutritional and bioactive components, in vitro protein digestibility, reducing power, metal chelating capacity, antioxidant activity, and anti-nutritional components of lima and adzuki beans. The findings showed that during the germination treatment, the in vitro protein digestibility of lima and adzuki beans increased by 14.75 and 10.98%, respectively. There was an increase in the antioxidant activity of lima beans by 33.48% and adzuki beans by 71.14% after 72 h of germination, respectively. The reducing power assay of lima and adzuki beans indicated an increase of 49.52 and 36.42%, respectively, during germination. Similarly, the flavonoid and metal chelating activity increased in lima and adzuki beans after 72 h of germination. In contrast, the anti-nutrients, such as phytic acid, tannin content, and trypsin inhibitor activity, decreased significantly p < 0.05 after 72 h of germination. These results are encouraging and allow for utilizing the flour obtained from the germinated beans in functional bakery products, which can contribute to eradicating protein deficiency among some population groups. At the same time, promoting soaking and germination of the beans as a way to enhance the nutritional quality and reduce anti-nutrients can contribute to the interest in these underutilized pulses. They could be seen as an additional tool to improve food security. Full article

Oat protein is unstable in intestinal fluid digestion, and it is easily degraded by trypsin and chymotrypsin, producing low molecular weight peptides. Endopeptidase hydrolysis can improve the bioavailability of active peptides and avoid further digestion in the gastrointestinal tract. Antimicrobial peptides (AMPs) can effectively improve host immunity, but most related studies focus on physiology and ecology, and there are few reports on their molecular level. Therefore, in this article, oat peptides were prepared via the simulated digestion method in vitro, and the main metabolites and action factors affecting colitis were screened by using the multi-omics methods in a high-throughput mode to analyze the effect and mechanism of colitis. Firstly, oat antimicrobial peptides were prepared from cationic resin combined with HPLC, and the anti-inflammatory effects of antimicrobial peptides were analyzed in vitro through the use of human colon epithelial (HCoEpiC) anti-inflammatory cells. In vivo experiments using rats have verified that AMPs can effectively prevent colitis caused by dextran sodium sulfate (DSS), reduce intestinal inflammatory cell infiltration and glandular disappearance in the colon, and reduce the apoptosis rate of colon cells. Secondly, metabolomics and transcriptomics were combined to analyze the mechanism of preventing enteritis, and it was found that oat antimicrobial peptides can promote DAG diglycerol production and inhibit the activation of T helper cells (TH), resulting in the down-regulation of key factors in the main downstream pathways of TH1, TH2 and TH17, and inhibit the production of inflammatory cells. At the same time, AMP can activate the wnt pathway, improve the expression of key genes of wnt and frizzled, promote the generation of intestinal stem cells, facilitate the differentiation and repair of intestinal epithelial cells, and prevent the generation of enteritis. Finally, the underlying genetic regulatory network of the important pathway was constructed from the effect of AMP on rat colitis. Full article

This research aimed to determine the biofunctional properties of wheat flour (WF) protein fractions and modifications to the antioxidant, anti-α-amylase and anti-angiotensin-I converting enzyme (ACE) activities induced by the action of digestive endopeptidases in vitro. A molecular characterization of the most abundant protein fractions, i.e., albumins, glutelins-1, glutelins-2 and prolamins, showed that low- and high-MW polypeptides rich in cysteine, glutamic acid and leucine were present in albumins and glutelins, whereas low-MW subunits with a high proportion of polar amino acids prevailed in prolamins. Prolamins exhibited the second-highest water holding capacity (54%) after WF (84%), while albumins provided superior foam stability (76%). Prolamins, glutenins-1 and globulins demonstrated the highest antioxidant activity (up to 95%, 68% and 59%, respectively) both before and after hydrolysis with pepsin (P-H) or trypsin–chymotrypsin (TC-H). Prolamins, globulins and WF strongly inhibited α-amylase (>90%) before and after TC-H, and before P-H (55–71%). Moreover, P-H significantly increased α-amylase inhibition by albumins from 53 to 74%. The fractions with strong ACE inhibitory activity (70–89%) included prolamins and globulins after TC-H or P-H, as well as globulins before TC-H and WF before P-H. This novel evidence indicates that WF protein fractions and their peptide-enriched P and TC hydrolysates are excellent sources of multifunctional bioactives with antioxidant, antihyperglycemic and antihypertensive potential. Full article