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Enrichment of staple foods with proteins can be a solution to tackle protein-energy malnutrition in the elderly. For instance, bread can be enriched with pea proteins that are cheap, sustainable and easily digestible. Non-conventional technologies, such as high pressure homogenization (HPH), can improve the digestibility of plant proteins. To characterize the health functionality of pea-enriched bread, a functional bread tailored to elderly consumers was developed by substituting 5% wheat flour with untreated or HPH-treated pea protein concentrate. Protein digestibility and colon fermentability were assessed by mimicking elderly in vitro gastrointestinal and gut microbiota conditions and compared with adult conditions. Bread reformulation with pea proteins affected physical and chemical properties and produced an increase in hardness, which is one of the key features for the acceptability of bread by the elderly. The highest hardness value was observed for pea protein bread, followed by HPH-treated pea protein bread and wheat bread. In vitro protein digestibility and fermentability were affected by reformulation and by physiological digestive conditions, with lower digestibility under elderly conditions compared to adult ones. The obtained results may contribute to a better understanding of food digestibility under different gastrointestinal conditions and its dependence on physiological and formulation factors, and ultimately would help to design age-tailored foods.

The effect of two processing methods of Jack beans ( i.e. cooked bean (CB) and cooked tempeh (CT)) on the in vitro digestibility of protein and starch, as well as the production of short chain fatty acids (SCFAs), γ-aminobutyric acid (GABA), and tryptophan (Trp) metabolites after in vitro colonic fermentation, was investigated. CT was obtained by fungal fermentation after cooking under acidic conditions. CT had significantly higher protein, lower digestible starch, lower total fiber, higher free phenolic compounds, and higher ash content compared to CB. CT exhibited better in vitro protein digestibility than CB and less glucose release during in vitro digestion than CB. A comparable concentration of total SCFAs and GABA was produced after in vitro fermentation of CB and CT, but CB produced more indole than CT, resulting in higher amounts of total Trp metabolites. In summary, our findings show that tempeh fermentation improves the nutritional quality of Jack beans and describe the impact of fermentation on the digestibility of nutrients and the formation of metabolites during colonic fermentation.

Lipids in almonds are naturally encapsulated by cell walls which may reduce the actual metabolizable energy content of almonds. Oral processing increases the accessibility of lipids to digestive enzymes by grinding the almond matrix. This study aimed to investigate the effect of adding accompanying foods (chocolate and iceberg lettuce) to almonds on oral processing behaviour, bolus properties and predicted lipid release. Natural chewing times of four almond model foods including one almond (1.3 g), four almonds (4.6 g), one almond with chocolate (4.3 g) and one almond with iceberg lettuce (4.3 g) were collected from n = 59 participants in duplicate. Expectorated boli at the moment of swallowing were characterized for number and mean area of almond bolus particles. Predicted lipid bioaccessibility was estimated using a previously validated model. At similar bite size (4.3-4.6 g), the addition of chocolate and iceberg lettuce to almonds significantly decreased ( p < 0.05) chewing time and significantly increased ( p < 0.05) eating rate compared to consumption of almonds alone. Almond bolus particle sizes were similar for almonds consumed alone (one and four almonds) and with chocolate, while consuming almonds with lettuce generated significantly fewer and larger almond bolus particles ( p < 0.05). Predicted lipid bioaccessibility of almonds consumed with iceberg lettuce was significantly lower ( p < 0.05) than for almonds consumed alone (one and four almonds) and almonds consumed with chocolate. Eating rate correlated significantly and positively with the mean area of bolus particles and significantly and negatively with predicted lipid release. In conclusion, combining almonds with other foods such as chocolate and lettuce influences oral processing behaviour and bolus properties and consequently predicted lipid bioaccessibility of almonds, highlighting the impact of food matrix and consumption context on these aspects.

This research assessed the influence of fermentation and germination as well as of particle size on lignan bioaccessibility from flaxseed by simulated in vitro gastrointestinal digestion. In vitro simulated colonic fermentation was used to study lignan release and its conversion into enterolignans. In addition, tea was included as a representative sample to investigate the stability of lignans in the gastrointestinal tract. Only secoisolariciresinol (SECO) was detected in flaxseed samples. SECO bioaccessibility in fermented flaxseed was highest among all matrices but limited to ≈1% ( P < 0.001). Lignan bioaccessibility was significantly influenced by particle size too ( P < 0.001 for both). In the colon, fermented flaxseed produced the highest SECO release among all flaxseed samples (≈65%), and the highest conversion of enterolignan (≈1.0%), whereas the conversion of lignans in tea brew was relatively high (≈15%). Lignan conversion varies greatly among donors due to inter-individual differences in microbiota activity. Food fermentation could be a viable strategy for increasing lignan release and conversion to enterolignan.

It has been recognized that, next to dietary fibre and proteins, gut microbiota can metabolize lipids producing bioactive metabolites. However, the metabolism of dietary lipids by human gut microbiota has been poorly explored so far. This study aimed to examine the change in lipids, particularly linoleic acid (LA), induced by the chemical form of lipids and the presence of the plant matrix. Short-chain fatty acid (SCFA) production was monitored to get an insight into microbial activity. Free LA, glyceryl trilinoleate and soybean oil as well as digested intact (DS) and broken (BS) soybean cells were subjected to in vitro fermentation using human faecal inoculums. Confocal microscopy was used to visualize the soybean cell integrity. Three LA metabolites, including two conjugated fatty acids (CLAs, 9z,11e and 9e,11e) and 12hydroxy, 9z C18:1, were identified and monitored. Free LA addition improved the LA metabolite production but reduced SCFA concentrations compared to trilinoleate and soybean oil. Breaking cell integrity had impacts on CLA, hydroxy C18:1 and SCFA production and free fatty acid release within the first 24 h of fermentation, but this effect vanished with time. In contrast, soybean oil only increased free LA release and hydroxy C18:1 production. The content of several FAs decreased during fermentation suggesting a substantial conversion in microbial metabolites. Besides, LA metabolites were also identified in the fermentation pellets suggesting the incorporation of microbial FA metabolites into bacterial cells. This study expands our understanding of microbial metabolism of dietary lipids with a special emphasis on the role of food- and diet-related factors.

Immature grain represents a precious nutritional source in many rural Africa areas. To optimize processing of immature rice into pepeta (a traditional rice-flakes produced from immature rice grains), immature rice (TXD306 variety) harvested at 18 and 26 days after 50% heading were processed in the laboratory under different soaking (0 and 12 h) and roasting temperature (80, 100 and 120 °C) regimes. Riboflavin, nicotinic acid, nicotinamide and iron concentration increased with severity of roasting temperature, while thiamine has an opposite trend. Heating promoted the transformation of insoluble into soluble dietary fiber, increased lipid digestibility decreasing protein one, which showed the highest value when rice was roasted at 100 °C. Soaking before roasting significantly increased moisture and iron content while slightly increased riboflavin, nicotinic acid and nicotinamide when compared to unsoaked products. Among roasted products, starch digestibility increased with roasting temperature. Microstructure analysis indicated a complete loss of cell wall integrity in cooked rice, determining a complete starch and protein digestion while this is delayed in raw rice and roasted products. We concluded that roasting at 100 °C is the optimum temperature to produce pepeta of the highest protein digestibility and low starch digestibility. Soaking before roasting at 120 °C is best when retaining micronutrients is considered.

In the present study we investigated the effect of cellular integrity on microbial utilization of proteins and carbohydrates by gut microbiota. Cotyledon cells from red kidney beans with different levels of structural integrity were fermented in-vitro by microbial communities previously adapted to the conditions of ascending, transverse and descending colon. The effect of bacterial adaptation to substrate was also assessed by using microbiota exposed to a diet rich in bean cells. Microscopy analyses indicate that cell integrity was maintained during fermentation. The amount of gas generated and the rate of total gas production was higher in broken cells compared to intact cells which suggest a faster and more extensive utilization of nutrients when cell wall is broken. A significantly higher butyric and propionic acid level was detected in broken cells at the end of the fermentation. Moreover, adapted bacterial communities were more efficient in fermenting bean cells where higher amounts of butyrate were produced in all colon regions independently of sample integrity. Bacterial communities of the distal colon appeared to be the most efficient in carbohydrate and protein fermentation as witnessed by the higher levels of gas, and short chain fatty acids. It was also found that cell integrity and adaptation to bean cells modulate the hierarchy of nutrient utilization, with non-starch polysaccharides preferred over starch and proteins by microbiota exposed to bean cells. Our results demonstrated that structural aspects of foods, such as cell integrity in plant tissues, may modulate nutrients utilization by gut microbiota.

Rice is commonly consumed as fully mature grain, but immature rice is considered to have better nutrient and technological properties. This is attributed to changes in content and profile of nutritional and functional compounds during maturation. This study assessed the effect of maturity on nutrient content of rice grains, and in vitro digestibility of starch and protein, for immature rice grains of TXD306 and Lawama varieties. The effect of processing of immature rice into so-called pepeta, traditionally produced from immature rice grains and widely consumed in Tanzania, was studied as well. The results showed reductions in lipid, protein, ash, thiamine, nicotinic acid, nicotinamide, and soluble and insoluble dietary fibre contents during rice grain development. However, no effect of maturity on in vitro starch and protein digestibility was observed. The contents of protein, ash, lipid, nicotinamide, iron, zinc, and total, soluble and insoluble dietary fibre were higher in pepeta from both varieties than in the corresponding rice grains. Protein digestibility of pepeta flour was 58.9% higher than that of cooked rice for variety TXD306, and 73.8% higher for Lawama. Differential scanning calorimetry indicated that starch of processed immature rice was completely gelatinized whereas its susceptibility to digestion in vitro was slightly lower than for cooked rice, possibly due to the higher cellular integrity retained after processing. These results demonstrate that pepeta-type processing improves the nutritional properties of rice and its potential use as a snack or ingredient in cereal-based formulas.

The inhibitory effect of tea polyphenols on starch digestibility can contribute to the control of the glycaemic index of starchy food. In this study, wheat bread and gluten-free bread were co-digested in vitro with different amounts of tea polyphenols. The kinetics of starch digestion and polyphenol bio-accessibility during in vitro digestion were monitored. The results showed that co-digestion of bread with tea polyphenols dose-dependently slowed the starch digestion kinetics and this effect is influenced by the types of polyphenols and the presence of gluten. The presence of gluten lowered the inhibitory efficacy of tannins on starch digestibility to 7.4% and 47.5% when 25 mg of tannins were co-digested with wheat bread and gluten-free bread, respectively. In contrast, the presence of gluten had little impact on the inhibitory efficacy of monomeric polyphenols. This study shows that the release of tea polyphenols in the digestive environment is a promising strategy for controlling the glycaemic index of starchy food and that monomeric and polymeric tea polyphenols differently affect starch digestion according to the presence of gluten.

Broccoli is rich in glucosinolates, which can be converted upon chewing and processing into Aryl hydrocarbon Receptor (AhR) ligands. Activation of AhR plays an important role in overall gut homeostasis but the role of broccoli processing on the generation of AhR ligands is still largely unknown. In this study, the effects of temperature, cooking method (steaming versus boiling), gastric pH and further digestion of broccoli on AhR activation were investigated in vitro and in ileostomy subjects. For the in vitro study, raw, steamed (t = 3 min and t = 6 min) and boiled (t = 3 min and t = 6 min) broccoli were digested in vitro with different gastric pH. In the in vivo ileostomy study, 8 subjects received a broccoli soup or a broccoli soup plus an exogenous myrosinase source. AhR activation was measured in both in vitro and in vivo samples by using HepG2-Lucia™ AhR reporter cells. Cooking broccoli reduced the AhR activation measured after gastric digestion in vitro, but no effect of gastric pH was found. Indole AhR ligands were not detected or detected at very low levels both after intestinal in vitro digestion and in the ileostomy patient samples, which resulted in no AhR activation. This suggests that the evaluation of the relevance of glucosinolates for AhR modulation in the gut cannot prescind from the way broccoli is processed, and that broccoli consumption does not necessarily produce substantial amounts of AhR ligands in the large intestine.

Apart from the presence of antinutritional factors, digestibility of soybean proteins is limited in intact cells by cell wall permeability to proteolitic enzymes. Food processing may modulate cell wall permeability and hence the accessibility of protease enzymes to intracellular proteins. In this study, soybeans were processed in various ways, e.g. cooking applied alone or with either germination or fermentation processes, and the modification in cell wall permeability was investigated using confocal microscopy to visualize the penetration of FITC-dextran probes into isolated cells/cell clusters. Diffusion of fluorescently labelled trypsin into cells and cell clusters was also monitored. Microscopy observations showed that fermentation and germination as well as proteolitic enzymes increase the permeability of boiled soybean cotyledon cells. The diffusion of trypsin into all the isolated cells was observed at an early stage of simulated in vitro digestion, whereas diffusion into cell clusters was delayed due to a bigger size and limited permeability of cell clusters. A modest, although significant, increase in protein digestibility was observed when boiling was combined with fermentation or germination likely due to pre-digestion of storage proteins and inactivation of trypsin inhibitors. This study highlights the positive role of fermentation and germination in improving protein digestibility in soybeans but overall suggests that cell wall permeability to trypsin plays a minor role in the extent of protein digestion of intact soybean cells.

Soybeans represent the largest source of plant proteins on the planet but their proteins are associated with low digestibility. Although several studies addressed the limiting factors affecting the rate and extent of soy protein digestion, the net effect of the food matrix, especially of an intact cell wall, has been poorly investigated so far. The purpose of the present study was to examine the relationship between the cell matrix and protein hydrolysis during simulated in vitro digestion of soybean particles of different sizes prepared from unheated and boiled cotyledons. In addition, intact cells were isolated from unheated and autoclaved cotyledons and then digested with and without lipase inhibitors to understand the impact of an intact cell wall and the presence of oil bodies on soybean protein digestibility. Protein digestibility was the highest in the particles prepared after boiling of previously milled cotyledons compared to particles of the same size obtained by milling previously cooked cotyledons as well as of uncooked cotyledons. Protein digestibility in isolated intact cells was lower than that of extracted proteins regardless of the thermal load applied whereas inhibition of pancreatic lipase reduces protein digestibility only slightly. The data indicated that the cell wall could contribute to limit protein digestion in soybean tissues; however, it is not an absolute barrier to pancreatic proteases. An accurate design of the milling and cooking process could be instrumental to modulate the digestion kinetics of soybean proteins.

Previous studies have proved that the physical encapsulation of nutrients by the cell walls of plant foods modulates macronutrient bioaccessibility during human digestion. In this study, we investigated structural factors that modulate lipid hydrolysis during in vitro digestion of raw and roasted hazelnut particles and isolated oil bodies. Isolated oil bodies exhibited a significantly higher lipid hydrolysis compared to hazelnut particles. Moreover, roasting had an impact on the structure of hazelnut cell walls implying a more efficient diffusion of digestive fluids and enzymes into the hazelnut cells. Heat treatment also caused destabilization of oil body interfacial protein membranes, facilitating their proteolysis under gastric conditions, altering the emulsion properties and enhancing fatty acid release during intestinal digestion. This study underlined the barrier role played by the plant cell wall as well as the impact of heat processing on lipid bioaccessibility in hazelnuts.