Identification, Bioaccessibility, and Antioxidant Properties of Phenolic Compounds in Carob Syrup.

Carob syrup is a brown, thick syrup produced from carob pulp that can be directly consumed or used as a sweetener, which also finds applications in folk medicinal practices. In this work, the quali-quantitative phenolic profile of five different carob syrups was elucidated before and after in vitro gastro-intestinal digestion. Moreover, the anti-oxidant properties of undigested and digested carob syrups were investigated. A total of 75 phenolic compounds were identified in undigested carob syrups. The most important phenolic compound in all the samples was gallic acid, the concentration of which ranged between 54.28 and 117.73 mg/100 g. Additional compounds belonging to the classes of hydroxybenzoic acids (in particular glycosylated gallic acid derivatives), hydroxycinnamic acids, and flavonoids (especially flavonols) were also identified. During in vitro gastric digestion, gallic acid mono- and di-hexosides were diglycosylated, releasing gallic acid, which was further degraded in ellagic acid through oxidative polymerization in the intestinal phase of the digestion. Ellagic acid was the major compound detected after in vitro gastro-intestinal digestion of carob syrups. With few exceptions, the anti-oxidant properties of carob syrup were preserved even after digestion. Carob syrup can be considered an important source of phenolic compounds with demonstrated positive effects on human health.

Antioxidant and Angiotensin I-Converting Enzyme (ACE) Inhibitory Peptides Obtained from Alcalase Protein Hydrolysate Fractions of Hemp (Cannabis sativa L.) Bran.

Proteins from hemp bran (HPB), a byproduct of the hemp seed food-processing chain, were chemically extracted, hydrolyzed by Alcalase, and separated by membrane ultrafiltration into four fractions (MW <1, 1-3, 3-5, and >5 kDa). The antioxidant and antihypertensive properties of the initial extract and the fractions were evaluated by in vitro assays for their ability to scavenge radical species, bind with metal ions, reduce ferric ions, and inhibit angiotensin-converting enzyme (ACE) activity. Bioactive peptides were identified by high-resolution mass spectrometry and sequence comparison with BIOPEP and BioPep DB databases. The hydrolysate was strongly antioxidant and ACE-inhibiting; the most bioactive peptides were further concentrated by ultrafiltration. Of the 239 peptides identified, 47 (12 antioxidant and 35 ACE-inhibitory) exhibited structural features correlated with the specific bioactivity. These results highlight the promise of hydrolysate and size-based HPB fractions as natural functional ingredients for the food or pharmaceutical industry.

Colonic In Vitro Model Assessment of the Prebiotic Potential of Bread Fortified with Polyphenols Rich Olive Fiber.

The use of olive pomace could represent an innovative and low-cost strategy to formulate healthier and value-added foods, and bakery products are good candidates for enrichment. In this work, we explored the prebiotic potential of bread enriched with Polyphenol Rich Fiber (PRF), a defatted olive pomace byproduct previously studied in the European Project H2020 EcoProlive. To this aim, after in vitro digestion, the PRF-enriched bread, its standard control, and fructo-oligosaccharides (FOS) underwent distal colonic fermentation using the in vitro colon model MICODE (multi-unit colon gut model). Sampling was done prior, over and after 24 h of fermentation, then metabolomic analysis by Solid Phase Micro Extraction Gas Chromatography Mass Spectrometry (SPME GCMS), 16S-rDNA genomic sequencing of colonic microbiota by MiSeq, and absolute quantification of main bacterial species by qPCR were performed. The results indicated that PRF-enriched bread generated positive effects on the host gut model: (i) surge in eubiosis; (ii) increased abundance of beneficial bacterial groups, such as Bifidobacteriaceae and Lactobacillales; (iii) production of certain bioactive metabolites, such as low organic fatty acids; (iv) reduction in detrimental compounds, such as skatole. Our study not only evidenced the prebiotic role of PRF-enriched bread, thereby paving the road for further use of olive by-products, but also highlighted the potential of the in vitro gut model MICODE in the critical evaluation of functionality of food prototypes as modulators of the gut microbiota.

Gluten free sourdough bread enriched with cricket flour for protein fortification: Antioxidant improvement and Volatilome characterization.

Insects represent a novel source of edible high nutritional value proteins which are gaining increasing interest as an alternative to traditional animal foods. In this work, cricket flour was used to produce gluten-free sourdough breads, suitable for celiac people and "source of proteins". The doughs were fermented by different methods and pH and microbial growth, volatile compounds, protein profile, and antioxidant activity, before and after baking, were analyzed and compared to standard gluten-free doughs. The results showed that cricket-enriched doughs and the standard had similar fermentation processes. Cricket enrichment conferred to the breads a typical flavoring profile, characterized by a unique bouquet of volatile compounds, made by nonanoic acid, 2,4-nonadienal (E,E), 1-hexanol, 1-heptanol, and 3-octen-2-one, expressed in different amounts depending on the type of inoculum. Finally, antioxidant activities were significantly enhanced in cricket breads, indicating that cricket powder provides to bakery gluten-free goods high nutritional value proteins and antioxidant properties.

Functional, nutritional, antioxidant, sensory properties and comparative peptidomic profile of faba bean (Vicia faba, L.) seed protein hydrolysates and fortified apple juice.

Enzymatic hydrolysis of plant-derived proteins can improve their quality by offering opportunities for food applications. In this study, three proteolytic enzymes (pepsin, trypsin, Alcalase®) were used, alone or combined, to produce faba bean protein hydrolysates (PHs). Their functional, nutritional and antioxidant properties were evaluated, and the peptidomic profile was assessed by LC-MS/MS. Hydrolysis improved solubility of faba proteins at acidic and neutral pH, and their antioxidant properties. Peptidomic analysis identified 2031 peptides in the different PHs. Among them, 9 showed 100% homology with previously known antioxidant peptides and several others had antioxidant motifs in their sequences. Sensory data analysis showed that after addition of PHs to apple juice, no significant differences were perceived between control and some of the PHs. This study demonstrates that enzymatic hydrolysis enhances the functional and antioxidant properties of faba bean proteins. Specifically, hydrolysates can be used as functional food ingredients to produce fortified beverages.

Olive oil by-product as functional ingredient in bakery products. Influence of processing and evaluation of biological effects.

Nowadays, the strong demand for adequate nutrition is accompanied by concern about environmental pollution and there is a considerable emphasis on the recovery and recycling of food by-products and wastes. In this study, we focused on the exploitation of olive pomace as functional ingredient in biscuits and bread. Standard and enriched bakery products were made using different flours and fermentation protocols. After characterization, they were in vitro digested and used for supplementation of intestinal cells (Caco-2), which underwent exogenous inflammation. The enrichment caused a significant increase in the phenolic content in all products, particularly in the sourdough fermented ones. Sourdough fermentation also increased tocol concentration. The increased concentration of bioactive molecules did not reflect the anti-inflammatory effect, which was modulated by the baking procedure. Conventionally fermented bread enriched with 4% pomace and sourdough fermented, not-enriched bread had the greatest anti-inflammatory effect, significantly reducing IL-8 secretion in Caco-2 cells. The cell metabolome was modified only after supplementation with sourdough fermented bread enriched with 4% pomace, probably due to the high concentration of tocopherol that acted synergistically with polyphenols. Our data highlight that changes in chemical composition cannot predict changes in functionality. It is conceivable that matrices (including enrichment) and processing differently modulated bioactive bioaccessibility, and consequently functionality.

Changes in carotenoids, phenolic acids and antioxidant capacity in bread wheat doughs fermented with different lactic acid bacteria strains.

Amongst the processing technologies able to improve the functional features of cereal-based foods, sourdough fermentation using Lactic Acid Bacteria (LAB) has been recently rediscovered for its beneficial effects. Wheat (Triticum aestivum L.) bread doughs were prepared using LAB strains belonging to different Lactobacillus species and changes in phenolic acid, carotenoid content and antioxidant capacity were evaluated. Two L. plantarum strains out of six were able to significantly increase carotenoid content in the dough, suggesting that a higher mobilization/solubilisation of these antioxidant compounds occurs. Within different fractions (free, soluble-conjugated, insoluble-bound), the relative distribution of ferulic acid and antioxidant activity changes depending on the specific strain. Overall, results indicate that some LAB strains cause in situ changes, significantly increasing the content of functional compounds in doughs during fermentation. This, in turn, could improve the functional features of bakery foods characterised by a high content in carotenoids and other bioactive compounds.

Sourdough Fermentation Favorably Influences Selenium Biotransformation and the Biological Effects of Flatbread.

Although selenium is of great importance for the human body, in several world regions the intake of this essential trace element does not meet the dietary reference values. To achieve optimal intake, fortification of bread by using selenium-enriched flour has been put forward. Less is known on the potential effect of sourdough fermentation, which might be worth exploring as the biological effects of selenium strongly depend on its chemical form and sourdough fermentation is known to cause transformations of nutrients and phytochemicals, including the conversion of inorganic selenium into organic selenocompounds. Here we investigated the bio transformation of selenium by sourdough fermentation in a typical Italian flatbread (piadina) made with standard (control) or selenium-enriched flour. The different piadina were submitted to in vitro digestion, and the biological activity of the resulting hydrolysates was tested by means of cultured human liver cells exposed to an exogenous oxidative stress. The use of selenium-enriched flour and sourdough fermentation increased the total content of bioaccessible selenium in organic form, compared to conventional fermentation, and led to protective effects counteracting oxidative damage in cultured cells. The present study suggests that selenium-rich, sourdough-fermented bakery products show promise for improving human selenium nutrition whenever necessary.

In Vivo Effects of Einkorn Wheat (Triticum monococcum) Bread on the Intestinal Microbiota, Metabolome, and on the Glycemic and Insulinemic Response in the Pig Model.

Einkorn wheat (Triticum monococcum) is characterized by high content of proteins, bioactive compounds, such as polyunsaturated fatty acids, fructans, tocols, carotenoids, alkylresorcinols, and phytosterols, and lower α-, ß-amylase and lipoxygenase activities compared to polyploid wheat. These features make einkorn flour a good candidate to provide healthier foods. In the present study, we investigated the effects of einkorn bread (EB) on the intestinal physiology and metabolism of the pig model by characterizing the glycemic and insulinemic response, and the microbiota and metabolome profiles. Sixteen commercial hybrid pigs were enrolled in the study; four pigs were used to characterize postprandial glycemic and insulinemic responses and twelve pigs underwent a 30-day dietary intervention to assess microbiota and metabolome changes after EB or standard wheat bread (WB) consumption. The postprandial insulin rise after an EB meal was characterized by a lower absolute level, and, as also observed for glucose, by a biphasic shape in contrast to that in response to a WB meal. The consumption of EB led to enrichment in short-chain fatty acid producers (e.g., Blautia, Faecalibacterium, and Oscillospira) in the gut microbiota and to higher metabolic diversity with lower content of succinate, probably related to improved absorption and therefore promoting intestinal gluconeogenesis. The observed changes, at both a compositional and metabolic scale, strongly suggest that EB consumption may support a health-promoting configuration of the intestinal ecosystem.

Olive oil industry by-products. Effects of a polyphenol-rich extract on the metabolome and response to inflammation in cultured intestinal cell.

Over the past years, researchers and food manufacturers have become increasingly interested in olive polyphenols due to the recognition of their biological properties and probable role in the prevention of various diseases such as inflammatory bowel disease. Olive pomace, one of the main by-products of olive oil production, is a potential low-cost, phenol-rich ingredient for the formulation of functional food. In this study, the aqueous extract of olive pomace was characterized and used to supplement human intestinal cell in culture (Caco-2). The effect on the cell metabolome and the anti-inflammatory potential were then evaluated. Modification in the metabolome induced by supplementation clearly evidenced a metabolic shift toward a "glucose saving/accumulation" strategy that could have a role in maintaining anorexigenic hormone secretion and could explain the reported appetite-suppressing effect of the administration of polyphenol-rich food. In both basal and inflamed condition, supplementation significantly reduced the secretion of the main pro-inflammatory cytokine, IL-8. Thus, our data confirm the therapeutic potential of polyphenols, and specifically of olive pomace in intestinal bowel diseases. Although intervention studies are needed to confirm the clinical significance of our findings, the herein reported results pave the road for exploitation of olive pomace in the formulation of new, value-added foods. In addition, the application of a foodomics approach allowed observing a not hypothesized modulation of glucose metabolism.

LC-ESI-QTOF-MS identification of novel antioxidant peptides obtained by enzymatic and microbial hydrolysis of vegetable proteins.

Bioactive antioxidant peptides are more and more attracting the attention of food manufacturers for their potential to transform food in functional food, able to prevent a variety of chronic diseases associated with oxidative stress. In the present study proteins extracted from different vegetable sources (KAMUT® khorasan wheat, emmer, lupine and pea) were hydrolyzed with commercial enzymes and Lactobacillus spp. strains. Hydrolysates were separated by size exclusion chromatography and purified fractions were analyzed for their antioxidant activity. Peptides from the fractions with the highest activity were identified by nanoLC-ESI-QTOF-MS and thirteen peptides were selected for synthesis on the basis of their sequence. Four peptides (VLPPQQQY, TVTSLDLPVLRW, VTSLDLPVLRW, FVPY) were found able to scavenge superoxide anion and hydroxyl radicals, organic nitro-radicals (ABTS, DPPH) and to inhibit lipid peroxidation. The impact of this work is targeted to add hydrolysed vegetable proteins to reformulated functional food or to produce health-promoting ingredients and nutraceuticals.

Bioavailability of Microencapsulated Iron from Fortified Bread Assessed Using Piglet Model.

The aim of this study was to investigate the influence of oral iron supplementation, in the form of fortified breads, on the growth performance, health, iron status parameters, and fecal metabolome of anemic piglets. A study was conducted on 24 hybrid (Large White × Landrace × Duroc) piglets. From day 44, the post-natal 12 piglets were supplemented with 100 g of one of two experimental breads, each fortified with 21 mg of ferrous sulphate, either encapsulated or not. After one week of oral supplementation, hematological parameters (hematocrit value, hemoglobin, and red blood cells) showed statistically significant differences (p ≤ 0.05). Piglets fed with the fortified breads had higher iron concentrations in the heart, liver, and intestinal mucosa compared to anemic piglets fed with control bread. Gene expression of hepcidin, iron exporter ferroportin (IREG1), and divalent metal transporter 1 (DMT1), together with concentrations of plasma ferritin, showed no significant statistical differences between groups. Both fortified breads could be used as sources of bioavailable iron. The seven-day intervention trial showed microencapsulation to have only a mild effect on the effectiveness of iron supplementation in the form of fortified bread.

Exploitation of starch industry liquid by-product to produce bioactive peptides from rice hydrolyzed proteins.

Small peptides show higher antioxidant capacity than native proteins and may be absorbed in the intestine without further digestion. In our study, a protein by-product from rice starch industry was hydrolyzed with commercial proteolytic enzymes (Alcalase, Neutrase, Flavourzyme) and microbial whole cells of Bacillus spp. and the released peptides were tested for antioxidant activity. Among enzymes, Alcalase was the most performing, while microbial proteolytic activity was less efficient. Conversely, the antioxidant activity was higher in the samples obtained by microbial hydrolysis and particularly with Bacillus pumilus AG1. The sequences of low molecular weight antioxidant peptides were determined and analyzed for aminoacidic composition. The results obtained so far suggest that the hydrolytic treatment of this industrial by-product, with selected enzymes and microbial systems, can allow its exploitation for the production of functional additives and supplements rich in antioxidant peptides, to be used in new food formulas for human consumption.